Research Article Lack of Association between Genetic...
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Research Article Lack of Association between Genetic Polymorphisms of JAK-STAT Signaling Pathway Genes and Acute Anterior Uveitis in Han Chinese Ling Cheng, Hongsong Yu, Yan Jiang, Juan He, Sisi Pu, Xin Li, and Li Zhang e First Affiliated Hospital of Chongqing Medical University, Chongqing Key Laboratory of Ophthalmology and Chongqing Eye Institute, Chongqing, China Correspondence should be addressed to Li Zhang; [email protected] Received 2 June 2016; Revised 14 September 2016; Accepted 16 October 2016 Academic Editor: Juan-Manuel Anaya Copyright © 2016 Ling Cheng et al. is is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Purpose. is study aimed to investigate the association between single nucleotide polymorphisms (SNPs) of JAK-STAT signaling pathway genes and acute anterior uveitis (AAU) with or without ankylosing spondylitis (AS) in the Han Chinese population. Methods. Eleven SNPs of the JAK1, JAK2, STAT1, IRF1, and NOS2 genes were analyzed in 443 AAU patients with AS, 486 AAU patients without AS, and 714 healthy controls. Genotyping was performed by PCR-RFLP assay or TaqMan probe assay. e Chi- squared ( 2 ) test and multivariate logistic regression analysis were used to compare the distributions of alleles and genotypes between patients and controls. values were adjusted using Bonferroni correction. Results. We did not observe significant differences in the genotype and allele frequencies of any SNP between AAU patients with or without AS and healthy controls. Stratification analyses by gender and HLA-B27 status showed a boundary significant association between two SNPs (rs10975003 and rs10758669) in JAK2 and AAU ( = 0.052 and = 0.053, resp.). Conclusions. Our results indicated that genetic polymorphisms of the JAK-STAT signaling pathway genes may not be associated with AAU in the Han Chinese population. 1. Introduction Uveitis is one of the major ocular diseases leading to blindness and visual impairment. e prevalence of uveitis is 111.3 per 100,000 persons in Taiwan [1] compared with 40.4 per 100,000 persons in Japan [2] and 115.3 per 100,000 persons in United States [3]. In the clinic, acute anterior uveitis (AAU), which may be accompanied by complicated phenotypes including cataract and glaucoma [4], is the most common type of uveitis [5]. Evidence suggests that the occurrence of AAU is associated with the prognosis of ankylosing spondylitis (AS) [6, 7]. e frequency of AAU, which is characterized by positive human leukocyte antigen- (HLA-) B27, varies across different ethnic populations [8–10]. In the United States and Western Europe, the prevalence of HLA- B27 with AAU is up to 50% [5, 8, 11]. Previous studies have reported that there is a strong association between AS and HLA-B27 in various ethnic groups [12–14]. Further study showed that the percentage of AAU accompanied by AS is 30–40%, suggesting that there may be linked pathogenesis between AAU and AS [15]. AAU and AS may share certain genetic associations, but several susceptibility genes seem to be unique for each disease [16]. Genes including TNFSF15, TRAF5, and FoxO1 have been reported to be associated with AAU [17–19]. However, a lack of association with AAU has been demonstrated for other genes, including CTLA4 and PTPN22 [20, 21]. A recent study revealed that T lymphocyte subsets (1 and 17) and CD4 + CD25 + Treg cells were involved in the development of HLA-B27 positive AAU [22, 23]. Furthermore, a higher level of 17 cells has been observed in the peripheral blood of patients with AS [24]. e Janus kinase-signal transducer and activator of tran- scription (JAK-STAT) signaling pathway plays a major role in T lymphocyte differentiation and function [25, 26]. JAK1 and JAK2 have been reported to play an important role in 1 and 17 cell differentiation [27]. STAT1 is critical to T lymphocyte differentiation and function [25, 26, 28]. STAT1 is activated by type I interferons (IFNs) and IFN- and plays Hindawi Publishing Corporation BioMed Research International Volume 2016, Article ID 5896906, 9 pages http://dx.doi.org/10.1155/2016/5896906
Transcript of Research Article Lack of Association between Genetic...
Research ArticleLack of Association between Genetic Polymorphisms ofJAK-STAT Signaling Pathway Genes and Acute AnteriorUveitis in Han Chinese
Ling Cheng Hongsong Yu Yan Jiang Juan He Sisi Pu Xin Li and Li Zhang
The First Affiliated Hospital of Chongqing Medical University Chongqing Key Laboratory of Ophthalmology andChongqing Eye Institute Chongqing China
Correspondence should be addressed to Li Zhang zhangli298hotmailcom
Received 2 June 2016 Revised 14 September 2016 Accepted 16 October 2016
Academic Editor Juan-Manuel Anaya
Copyright copy 2016 Ling Cheng et al This is an open access article distributed under the Creative Commons Attribution Licensewhich permits unrestricted use distribution and reproduction in any medium provided the original work is properly cited
PurposeThis study aimed to investigate the association between single nucleotide polymorphisms (SNPs) of JAK-STAT signalingpathway genes and acute anterior uveitis (AAU) with or without ankylosing spondylitis (AS) in the Han Chinese populationMethods Eleven SNPs of the JAK1 JAK2 STAT1 IRF1 and NOS2 genes were analyzed in 443 AAU patients with AS 486 AAUpatients without AS and 714 healthy controls Genotyping was performed by PCR-RFLP assay or TaqMan probe assay The Chi-squared (1205942) test and multivariate logistic regression analysis were used to compare the distributions of alleles and genotypesbetween patients and controls 119875 values were adjusted using Bonferroni correction Results We did not observe significantdifferences in the genotype and allele frequencies of any SNP between AAU patients with or without AS and healthy controlsStratification analyses by gender and HLA-B27 status showed a boundary significant association between two SNPs (rs10975003and rs10758669) in JAK2 and AAU (119875 = 0052 and119875 = 0053 resp)Conclusions Our results indicated that genetic polymorphismsof the JAK-STAT signaling pathway genes may not be associated with AAU in the Han Chinese population
1 Introduction
Uveitis is one of themajor ocular diseases leading to blindnessand visual impairment The prevalence of uveitis is 1113per 100000 persons in Taiwan [1] compared with 404 per100000 persons in Japan [2] and 1153 per 100000 persons inUnited States [3] In the clinic acute anterior uveitis (AAU)which may be accompanied by complicated phenotypesincluding cataract and glaucoma [4] is the most commontype of uveitis [5] Evidence suggests that the occurrenceof AAU is associated with the prognosis of ankylosingspondylitis (AS) [6 7] The frequency of AAU which ischaracterized by positive human leukocyte antigen- (HLA-)B27 varies across different ethnic populations [8ndash10] In theUnited States and Western Europe the prevalence of HLA-B27 with AAU is up to 50 [5 8 11] Previous studies havereported that there is a strong association between AS andHLA-B27 in various ethnic groups [12ndash14] Further studyshowed that the percentage of AAU accompanied by AS is
30ndash40 suggesting that there may be linked pathogenesisbetween AAU and AS [15] AAU and AS may share certaingenetic associations but several susceptibility genes seem tobe unique for each disease [16] Genes including TNFSF15TRAF5 and FoxO1 have been reported to be associated withAAU [17ndash19] However a lack of association with AAU hasbeen demonstrated for other genes including CTLA4 andPTPN22 [20 21] A recent study revealed that T lymphocytesubsets (Th1 and Th17) and CD4+ CD25+ Treg cells wereinvolved in the development of HLA-B27 positive AAU[22 23] Furthermore a higher level of Th17 cells has beenobserved in the peripheral blood of patients with AS [24]
The Janus kinase-signal transducer and activator of tran-scription (JAK-STAT) signaling pathway plays a major rolein T lymphocyte differentiation and function [25 26] JAK1and JAK2 have been reported to play an important role inTh1 and Th17 cell differentiation [27] STAT1 is critical to Tlymphocyte differentiation and function [25 26 28] STAT1is activated by type I interferons (IFNs) and IFN-120574 and plays
Hindawi Publishing CorporationBioMed Research InternationalVolume 2016 Article ID 5896906 9 pageshttpdxdoiorg10115520165896906
2 BioMed Research International
an important role in immune responses [29] IRF1 is the firstmember identified in the IRF family and is involved in manyinnate and adaptive immune responses [30] Impaired orabsent Th1-type immune responses favor Th2 differentiationin IRF1-deficientmice [31 32]NOS2-derivedNO a key factorin immunoregulation [33] can inhibit Th1 as well as Th2cytokine production and regulate the development of FoxP3+Treg cells [34 35] In summary JAK-STAT signaling pathwaygenes including JAK1 JAK2 STAT1 IRF1 and NOS2 havebeen suggested to be strongly linked with T cells and may beinvolved in the pathophysiology of AAU with or without AS
Thus we conducted the present case-control study toinvestigate whether JAK-STAT signaling pathway genes con-fer susceptibility to AAU risk in a Chinese Han population
2 Materials and Methods
21 Subjects A total of 929 AAU patients were enrolled inthis study including 443 patients with AS (AAU+AS+) and486 patients without AS (AAU+ASminus) as well as 714 gender-and race-matched healthy controls All subjects were HanChinese recruited from the Department of Ophthalmologyin the First Affiliated Hospital of Chongqing Medical Uni-versity (Chongqing China) between June 2008 and May2015 All AAU patients were diagnosed based on medicalrecords physical examinations and the anatomic locationof inflammation as previously described by Jabs et al [36]The diagnosis of AS followed the modified New York Criteria[37] All subjects gave written informed consent before bloodcollection This study was approved by the Human EthicsCommittee of the First Affiliated Hospital of ChongqingMedical University (Approval number 2009-201008) andfollowed the tenets of the Declaration of Helsinki
22 SNP Selection We selected candidate single nucleotidepolymorphisms (SNPs) based on previously published stud-ies and included only those SNPs significantly associatedwith autoimmune diseases [38ndash45] We used HaploView 42software to evaluate the linkage disequilibrium (LD) andminor allele frequency (MAF) of the SNPs Five SNPs ofJAK1 rs2780815 rs3790532 rs310230 rs310236 and rs310241[41 42] were selected Since the SNPs rs3790532 rs310230rs310236 and rs310241 are in strong LD with each other (1199032 gt08 Figure 1) we only used rs310241 in our study Further-more we also eliminated SNPs that were not polymorphicin the Chinese population Finally eleven SNPs in five JAK-STAT signaling pathway genes were tested in our studyincluding two SNPs in the intron region of the JAK1 gene(rs310241 rs2780815) [41] two SNPs in the exon region and31015840UTR of the JAK2 gene (rs10758669 rs10975003) [38 45]one SNP in the intron region of the IRF1 gene (rs2070721)four SNPs in the exon region and intron region of the STAT1gene (rs2066802 rs1547550 rs6718902 and rs10199181) [3940] and two SNPs in the exon region and intron region ofthe NOS2 gene (rs2297518 rs4795067) [43 44]
23 DNA Extraction and Genotyping Peripheral blood sam-ples were collected from subjects and genomic DNA extrac-tion was performed using the QIAamp DNA Blood Mini
84
84 83
83
11
11
10
3 11 12 71 83
rs2780815
rs310241
rs3790532
rs310236
rs310230
10
Block 1 (37kb)
Figure 1 Linkage disequilibrium (LD) analysis of rs2780815rs310241 rs3790532 rs310236 and rs310230 in the JAK1 gene TheLD block was estimated by HaploView software version 42 usingthe Chinese Han HapMap data The number in the square indicatesthe 1199032 value
Kit (Qiagen Valencia CA USA) The genomic DNA wasquantified with NanoDrop 2000 (Thermal Fisher Scien-tific Delaware USA) and stored at minus20∘C until use ThreeSNPs (rs2780815 rs2070721 and rs10199181) were genotypedby TaqMan probe (Applied Biosystems Foster City CA)and the others were genotyped by polymerase chain reac-tion restriction fragment length polymorphism (PCR-RFLP)assay The specific primers for PCR and restriction enzymesare described in Table 1 The PCR reactions were performedunder the following conditions denaturation at 95∘C for5 minutes 33 to 36 cycles of denaturation at 95∘C for 30seconds annealing at 56ndash64∘C for 30 s and extension at 72∘Cfor 30 seconds and a final extension at 72∘C for 5 minutesThe enzyme-digested products were visualized on 3 or4 agarose gels and stained with GoldView (SBS GenetechBeijing China) Direct sequencing was carried out randomlyon 10 of the study samples to assure the validity of theSNP genotyping method used The success rate of all SNPgenotyping ranged from 973 to 100
24 StatisticalAnalysis Hardy-Weinberg equilibrium (HWE)was analyzed by the 1205942 test The distributions of the alleleand genotype frequencies between the patients and controlswere compared by the 1205942 test Multivariate logistic regressionmodel adjusted for age and gender was further adopted totest the associations between the SNPs and AAU The riskeffect of each SNP was measured by odds ratios (OR) and95confidence intervals (CI)Therewere four differentmod-els of inheritance in our study including additive modelscodominantmodels dominantmodels and recessivemodels119875 values were corrected using the Bonferroni correction
BioMed Research International 3
Table 1 PCR primer sequences and restriction enzymes
SNP Primers Restriction enzyme
rs310241 51015840 AACCACCAGCTCAACATTCCTAG 3101584051015840 CAGCCAGGTCTCCCGTAGG 31015840 BseDI
rs6718902 51015840 CGGACAAAAGCATGCACTAGA 3101584051015840 CCACCACCATTAATAGGTGACTTTA 31015840 DraI
rs2297518 51015840 TGAGCTCTTTCAGCATGAAGATC 3101584051015840 CTTCCGTGGTGGGCTGTG 31015840 TaqI
rs10758669 51015840 TGATGTAGAGACAAGGACATGCTGAGGTAC 3101584051015840 GCCAAAAGACAAAGGCAAGGGG 31015840 BanI
rs10975003 51015840 GGCCAGTCAAGAAAAAACCAGTT 3101584051015840 TTCGGAGTCTTGTCTGAGCATGT 31015840 HpaI
rs4795067 51015840 GCACTCATTCATTCATGCAAACATA 3101584051015840 GGCAGAACTTGAACCCCAGCT 31015840 NdeI
rs1547550 51015840 CTTCTCTAGGAGGCCCAGCA 3101584051015840 TGGGCACCACGATATGAGAG 31015840 BstMAI
rs2066802 51015840 ATTCCTGGAGCAGGTTCACAAG 3101584051015840 AAACATGGCCCCAAGTCACT 31015840 HindIII
method considering multiple tests Statistical significancelevel was defined as a corrected 119875 value lt 005 Statisticalanalyses were performed using SPSS version 170 (SPSS IncChicago IL USA)
3 Results
31 Clinical Features of AAU Patients The detailed clinicalfeatures and demographic characteristics of the AAUpatientsare presented in Table 2 All 929 AAU patients include 569(612) males and 360 (388) females The 714 controlsubjects consisted of 428 (599) males and 286 (401)females The average age was 398 plusmn 123 in AAU patientsand 395 plusmn 108 in the controls respectively There were nosignificant differences in age and gender between the casesand controls In addition 546 AAU patients (689) wereHLA-B27-positive whereas 246 AAU patients (311) wereHLA-B27-negative
32 The Genotype and Allele Frequency Distribution of theTested SNPs in AAU Eleven SNPs of the JAK-STAT signalingpathway genes (JAK1 JAK2 STAT1 IRF1 and NOS2) weresuccessfully genotyped There were no significant deviationsof HWE in either the cases or controls We did not observesignificant differences in the genotype and allele distribu-tions of any of the SNPs between the AAU patients andcontrol subjects after Bonferroni correction (see Supplemen-tary Table S1 in Supplementary Material available onlineat httpdxdoiorg10115520165896906) Further stratifiedanalyses of gender AS andHLA-B27 status showed a bound-ary significant association of two SNPs (rs10975003 andrs10758669) of JAK2 with AAU In female AS-positive AAUpatients there was a decreased frequency of the TT genotypeof rs10975003 compared to the female controls (OR = 055119875 = 159 times 10minus3 119875Bonferroni = 0052 Table 3) whereas nosignificant differences were observed in the genotype andallele frequencies of the other ten SNPs (119875Bonferroni gt 005Table 3) Similarly there were no significant differences in
Table 2 Clinical characteristics of the investigated subjects
Clinical features Total PercentageAAU patients 929 100Mean age plusmn SD (years) 398 plusmn 123AAU with AS 443 477AAU without AS 486 523AAU male 569 612AAU female 360 388AAU with AS (male) 326 (443 tested) 736AAU with AS (female) 117 (443 tested) 264AAU without AS (male) 243 (486 tested) 50AAU without AS (female) 243 (486 tested) 50HLA-B27+ AAU 546 (792 tested) 689HLA-B27+ AAU AS+ 348 (428 tested) 813HLA-B27+ AAU ASminus 198 (364 tested) 544Control 714 100Mean age plusmn SD (years) 395 plusmn 108Male 428 599Female 286 401
the genotype and allele frequencies of the SNPs between themale AAU patients and male controls (119875Bonferroni gt 005Supplementary Table S2)
In addition an increased frequency of theAC genotype inrs10758669 was observed in HLA-B27-positive AAU patientscompared to the healthy controls (OR = 144119875 = 162times10minus3119875Bonferroni = 0053 Table 4) whereas there were no significantdifferences in the genotype and allele frequencies of the otherten SNPs between the HLA-B27-positive AAU patients andthe control subjects (119875Bonferroni gt 005 Table 4)
However an increased frequency of the rs10758669ACgenotype was observed in HLA-B27-positive AS-positiveAAU patients compared to healthy controls (OR = 149 119875 =256times10minus3119875Bonferroni = 0084 Table 5) whereas no significant
4 BioMed Research InternationalTa
ble3Alleleandgeno
type
frequ
encies
infemaleA
AUpatie
ntsa
ndfemalec
ontro
ls
Gene
SNP
Alleleandgeno
type
AAU+AS+
(female)
AAU+ASminus
(female)
Con
trols(fe
male)119875(A
S+)
Pc(A
S+)
OR(95
CI)119875(A
Sminus)
Pc(A
Sminus)
OR(95
CI)
JAK1
rs310241
C80
(333
)138(296
)
156(287
)019
NS
124(090ndash
172)
074
NS
105(080ndash
137)
CC12
(100
)14
(60)
17(62)
019
NS
167(077ndash361)
091
NS
096
(046ndash
200)
CT56
(467
)110(472
)
122(449
)074
NS
108(070ndash
166)
060
NS
110(077ndash15
6)TT
52(433
)109(468
)133(489
)031
NS
080
(052
ndash123)
064
NS
092
(065ndash13
0)
rs2780815
G159(787
)40
0(870
)
495(853
)003
NS
064
(042ndash096)
045
NS
115(080ndash
163)
GG
63(624
)175(761
)212(731
)004
NS
061
(038ndash
099)
044
NS
117(079ndash
175)
GT
33(326
)50
(217
)
71(245
)011
NS
150(091ndash245)
046
NS
086
(057ndash12
9)TT
5(50)
5(22)
7(24)
020
NS
211(065ndash680)
086
NS
090
(028ndash287)
JAK2
rs10758669
A170(739
)328(675
)
399(723
)064
NS
109(077ndash15
4)010
NS
080
(061ndash10
4)AA
59(513
)
104(428
)144(522
)088
NS
097
(063ndash14
9)003
NS
069
(049ndash
100)
AC52
(452
)120(494
)
111(40
2)
036
NS
123(079ndash
190)
004
NS
145(102ndash205)
CC4(35)
19(78
)21
(76
)013
NS
044
(015
ndash130)
093
NS
103(054ndash
197)
rs10975003
C44
(206
)117
(258
)93
(179
)
040
NS
119(080ndash
177)
283times10minus3
009
159(117ndash217
)CC
2(19)
11(48)
8(31
)052
NS
060
(013
ndash287)
032
NS
160(063ndash406)
CT40
(374
)
95(419
)
77(296
)
015
NS
142(088ndash228)
483times10minus3
NS
171(118ndash249)
TT65
(607
)121(533)
175(673
)
023
NS
075
(047ndash12
0)15
9times10minus3
0052
055
(038ndash
080)
STAT
1
rs1547550
C205(869
)411(886)
423(870
)
095
NS
099
(062ndash15
6)047
NS
116(078ndash17
1)CC
88(746
)182(784
)185(761
)075
NS
092
(055ndash15
3)055
NS
114(074
ndash175)
CG29
(246
)47
(203
)53
(218
)
056
NS
117(070ndash
196)
068
NS
091
(059ndash
142)
GG
1(08
)3(13)
5(21
)040
NS
041
(005ndash352)
056
NS
065
(015
ndash276
)
rs2066
802
C46
(204
)81
(174
)
113(212
)
080
NS
095
(065ndash14
0)013
NS
078
(057ndash10
8)CC
1(09
)6(26)
8(30)
022
NS
029
(004ndash
234)
078
NS
086
(029ndash
250)
CT44
(389
)69
(296
)
97(363
)063
NS
112(071ndash17
6)011
NS
074
(051ndash10
7)TT
68(602
)158(678
)
162(607
)093
NS
098
(063ndash15
4)010
NS
137(095ndash19
7)
rs6718902
C132(600
)259(542
)308(538
)012
NS
129(094ndash
176)
091
NS
101(079ndash129)
CC36
(327
)72
(301
)82
(287
)043
NS
121(075ndash194)
072
NS
107(074
ndash156)
CT60
(545
)115(481
)144(503
)045
NS
118(076
ndash184)
061
NS
092
(065ndash12
9)TT
14(127
)52
(218
)
60(210
)
006
NS
055
(029ndash
103)
083
NS
105(069ndash
159)
rs10199181
A51
(248
)117
(261
)148(252
)091
NS
098
(068ndash14
1)073
NS
105(079ndash
139)
AA
6(58)
20(89)
19(65)
082
NS
090
(035ndash231)
029
NS
142(074
ndash273)
AT39
(379
)
77(344
)110(374
)
094
NS
102(064ndash
162)
048
NS
088
(061ndash12
6)TT
58(563
)127(567
)165(561
)097
NS
101(064
ndash158)
090
NS
102(072ndash14
5)
IRF1
rs2070721
A86
(406
)154(339
)216(343
)010
NS
131(095ndash180)
090
NS
098
(076
ndash127)
AA
15(14
2)
26(115
)
29(92)
015
NS
163(084ndash
317)
039
NS
128(073ndash223)
AC56
(528
)102(449
)158(502
)063
NS
111(072ndash173)
023
NS
081
(058ndash114)
CC35
(330
)99
(436
)128(406
)016
NS
072
(045ndash114)
049
NS
113(080ndash
160)
NOS2
rs2297518
A52
(205
)69
(152
)74
(157
)010
NS
139(094ndash
205)
084
NS
096
(068ndash13
8)AA
3(24)
6(26)
2(08)
024
NS
283
(047ndash1717)
014
NS
318
(063ndash1590)
AG46
(362
)57
(252
)70
(297
)
020
NS
135(085ndash213)
027
NS
080
(053
ndash120)
GG
78(614
)
164(722
)164(695
)
012
NS
070
(045ndash110)
051
NS
114(077ndash17
1)
rs4795067
A173(739
)369(759
)439(778
)
024
NS
081
(057ndash115)
046
NS
090
(067ndash12
0)AA
62(530
)145(597
)
169(599
)
020
NS
075
(049ndash
116)
095
NS
099
(070ndash
140)
AG49
(419
)
79(325
)101(358)
026
NS
129(083ndash201)
043
NS
086
(060ndash
124)
GG
6(51
)19
(78
)12
(43)
070
NS
122(045ndash332)
008
NS
191(091ndash4
02)
OR=od
dsratio
95
CI=95
confi
denceinterval
Pc=119875valuea
djustedby
Bonferroni
correctio
n
BioMed Research International 5
Table 4 Allele and genotype frequencies of SNPs in patients with AAU versus control subjects stratified by HLA-B27 status
Gene SNP Allele and genotype AAUHLA-B27 Control 119875 Pc OR (95 CI)
JAK1
rs310241
C 324 (310) 392 (275) 006 NS 118 (100ndash141)CC 41 (79) 45 (63) 029 NS 127 (082ndash196)CT 242 (464) 302 (424) 016 NS 118 (094ndash148)TT 239 (457) 366 (513) 005 NS 080 (064ndash100)
rs2780815
G 878 (862) 1250 (875) 035 NS 089 (070ndash113)GG 381 (748) 548 (767) 044 NS 090 (069ndash118)GT 116 (228) 154 (216) 061 NS 107 (082ndash141)TT 12 (24) 12 (17) 040 NS 141 (063ndash317)
JAK2
rs10758669
A 705 (659) 980 (690) 010 NS 087 (073ndash103)AA 220 (411) 346 (487) 008 NS 074 (059ndash092)AC 265 (495) 288 (406) 162 times 10minus3 0053 144 (115ndash180)CC 50 (94) 76 (107) 043 NS 086 (059ndash125)
rs10975003
C 251 (245) 272 (212) 005 NS 121 (100ndash147)CC 26 (51) 28 (44) 056 NS 118 (068ndash203)CT 199 (388) 216 (335) 006 NS 126 (099ndash160)TT 287 (561) 399 (621) 003 NS 077 (061ndash098)
STAT1
rs1547550
C 937 (872) 1254 (878) 067 NS 095 (075ndash121)CC 407 (758) 555 (777) 042 NS 090 (069ndash117)CG 123 (229) 144 (202) 024 NS 118 (090ndash154)GG 7 (13) 15 (21) 029 NS 062 (025ndash152)
rs2066802
C 190 (193) 288 (207) 039 NS 091 (074ndash112)CC 15 (30) 27 (39) 044 NS 078 (041ndash148)CT 160 (325) 234 (337) 066 NS 095 (074ndash121)TT 318 (645) 434 (624) 047 NS 109 (086ndash139)
rs6718902
C 584 (565) 778 (545) 033 NS 108 (092ndash127)CC 163 (315) 209 (293) 040 NS 111 (087ndash142)CT 258 (499) 360 (504) 086 NS 098 (078ndash123)TT 96 (186) 145 (203) 045 NS 090 (067ndash119)
rs10199181
A 270 (272) 397 (278) 075 NS 097 (081ndash116)AA 43 (87) 55 (77) 055 NS 114 (075ndash173)AT 184 (371) 287 (402) 028 NS 088 (069ndash111)TT 269 (542) 372 (521) 047 NS 109 (087ndash137)
IRF1 rs2070721
A 356 (357) 487 (341) 040 NS 108(091ndash127)AA 61 (122) 74 (104) 030 NS 121 (084ndash173)AC 234 (470) 339 (474) 087 NS 098 (078ndash123)CC 203 (408) 301 (422) 063 NS 094 (075ndash119)
NOS2
rs2297518
A 194 (181) 209 (162) 021 NS 115 (093ndash142)AA 12 (23) 12 (19) 064 NS 121 (054ndash272)AG 170 (317) 185 (286) 024 NS 116 (090ndash149)GG 354 (660) 450 (695) 020 NS 085 (067ndash109)
rs4795067
A 797 (743) 1008 (761) 035 NS 092 (076ndash110)AA 293 (546) 382 (577) 033 NS 089 (071ndash123)AG 211 (394) 244 (369) 037 NS 111 (088ndash141)GG 32 (60) 36 (54) 069 NS 110 (068ndash180)
OR = odds ratio 95 CI = 95 confidence intervalPc = 119875 value adjusted by Bonferroni correction
6 BioMed Research InternationalTa
ble5Alleleandgeno
type
frequ
encies
ofSN
Psin
patientsw
ithAAU
versus
controlsub
jectsstratified
byASandHLA
-B27
status
Gene
SNP
Alleleandgeno
type
AAU+AS+
HLA
-B27
AAU+ASminus
HLA
-B27
Con
trol
119875(A
S+)
Pc(A
S+)
OR(95
CI)119875(A
Sminus)
Pc(A
Sminus)
OR(95
CI)
JAK1
rs310241
C205(316
)
119(299
)
392(275
)
005
NS
122(100ndash
149)
032
NS
113(088ndash14
5)CC
30(93)
11(55)
45(63)
009
NS
152(094ndash
245)
070
NS
087
(044ndash
172)
CT145(447
)97
(487
)302(424
)047
NS
110(085ndash14
4)010
NS
131(095ndash179)
TT149(460
)90
(457
)366(513
)
011
NS
081
(062ndash10
5)014
NS
079
(058ndash10
8)
rs2780815
G543(843
)335(896
)
1250
(875
)
005
NS
077
(059ndash
100)
028
NS
122(085ndash17
7)GG
230(714
)
151(807)
548(767
)007
NS
076
(056ndash
102)
024
NS
127(085ndash19
0)GT
83(258
)33
(176
)
154(216
)
014
NS
126(093ndash17
2)023
NS
078
(051ndash118)
TT9(28)
3(17)
12(17)
024
NS
168(070ndash
403)
094
NS
095
(027ndash342)
JAK2
rs10758669
A44
9(662
)256(653
)980(690
)
020
NS
088
(072ndash17
0)016
NS
085
(067ndash10
7)AA
139(410
)
81(413
)
346(487
)002
NS
073
(056ndash
095)
007
NS
074
(054ndash
102)
AC171(504)
94(480
)288(406
)256times10minus3
0084
149(115
ndash194)
006
NS
135(098ndash18
6)CC
29(86)
21(107
)76
(107
)028
NS
078
(050ndash
122)
100
NS
100(060ndash
167)
rs10975003
C165(257
)86
(225
)272(212
)
003
NS
129(103ndash16
1)057
NS
108(082ndash14
3)CC
18(56)
8(42)
28(44)
039
NS
131(071ndash240)
092
NS
096
(043ndash214)
CT129(402
)70
(366
)216(335
)004
NS
133(101ndash17
5)044
NS
114(082ndash16
0)TT
174(542
)113(592
)
399(621
)001
NS
072
(055ndash095)
047
NS
089
(064ndash
123)
STAT
1
rs1547550
C608(881
)329(857
)1254
(878
)
084
NS
103(078ndash13
6)026
NS
083
(060ndash
115)
CC264(765
)143(745
)555(777
)
066
NS
093
(069ndash
127)
034
NS
084
(058ndash12
1)CG
80(232
)43
(224
)144(202
)026
NS
120(088ndash16
3)050
NS
114(078ndash16
8)GG
1(03
)6(31
)15
(21
)002
NS
014
(002ndash103)
040
NS
150(058ndash393)
rs2066
802
C130(207
)60
(168
)288(207
)099
NS
100(079ndash
126)
009
NS
077
(057ndash10
5)CC
11(35)
4(22)
27(39)
077
NS
090
(044ndash
183)
029
NS
057
(020ndash
164)
CT108(344
)52
(291
)
234(337
)082
NS
103(078ndash13
7)024
NS
081
(056ndash
116)
TT195(621
)123(687
)434(624
)092
NS
099
(075ndash13
0)012
NS
132(093ndash18
8)
rs6718902
C368(577
)
216(545
)778(545
)018
NS
114(094ndash
138)
098
NS
100(080ndash
125)
CC103(323
)60
(303
)209(293
)
033
NS
115(087ndash15
3)078
NS
105(075ndash14
8)CT
162(508
)96
(485
)360(504
)091
NS
102(078ndash13
2)063
NS
093
(068ndash12
7)TT
54(169
)42
(212
)
145(203
)020
NS
080
(057ndash113)
078
NS
106(072ndash15
6)
rs10199181
A170(269
)100(278
)
397(278
)
067
NS
096
(077ndash118)
099
NS
100(077ndash12
9)AA
25(79
)18
(100
)55
(77
)091
NS
103(063ndash16
8)032
NS
133(076
ndash233
)AT
120(380
)64
(356
)287(402
)050
NS
091
(069ndash
120)
026
NS
082
(058ndash115)
TT171(541
)98
(544
)372(521
)055
NS
108(083ndash14
1)057
NS
110(079ndash
153)
IRF1
rs2070721
A221(349)
135(373
)
487(341
)073
NS
103(085ndash12
6)025
NS
115(090-14
6)AA
37(117
)
24(133
)74
(104
)053
NS
114(075ndash174)
026
NS
132(081ndash216)
AC147(464
)87
(481
)339(474
)
074
NS
096
(073ndash12
5)089
NS
102(074
ndash142)
CC133(420
)70
(386
)301(422)
095
NS
099
(076
ndash130)
040
NS
097
(062ndash12
1)
NOS2
rs2297518
A130(19
0)
64(165
)209(162
)011
NS
122(096ndash
155)
087
NS
103(076
ndash139
)AA
7(20)
5(26)
12(19)
083
NS
111(043ndash284)
053
NS
140(049ndash
402)
AG116(339
)54
(278
)
185(286
)008
NS
128(097ndash17
0)084
NS
096
(067ndash13
8)GG
219(641
)135(696
)
450(695
)
007
NS
078
(059ndash
103)
099
NS
100(071ndash14
2)
rs4795067
A512(736
)295(745
)1008
(761
)020
NS
087
(071ndash10
8)061
NS
092
(071ndash119)
AA
184(529
)109(551
)382(577
)
014
NS
082
(063ndash10
7)051
NS
090
(065ndash12
4)AG
144(414
)
77(388
)244(369
)016
NS
121(093ndash158)
060
NS
109(079ndash
151)
GG
20(57)
12(61
)36
(54)
084
NS
106(060ndash
186)
074
NS
112(057ndash220)
OR=od
dsratio
95
CI=95
confi
denceinterval
Pc=119875valuea
djustedby
Bonferroni
correctio
n
BioMed Research International 7
differences in the genotype and allele frequencies of the other10 SNPs were observed between the HLA-B27-positive AS-positiveAAUpatients and healthy controls (119875Bonferroni gt 005Table 5) In addition there were no significant differencesin the genotype and allele frequencies of the tested SNPsbetween the AS-positive AAU patients and control subjects(119875Bonferroni gt 005 Supplementary Table S3)
33 Logistic Regression Analysis of SNPs in AAU We furtherinvestigated the SNPs rs10758669 and rs10975003 using addi-tive codominant dominant and recessive genetic modelsusing a multivariate logistic regression model adjusted forage and gender We observed that the frequency of the CAgenotype of rs10758669 was significantly higher in AAUpatients which suggests that patients with the rs10758669CA genotype have increased susceptibility to AAU (468versus 406 OR = 128 119875 = 002 SupplementaryTable S4) An increased frequency of the AC genotype ofrs10758669 was also observed in HLA-B27-positive AAUpatients and AS-positive AAU patients compared to thehealthy controls (495 versus 406 OR = 143 119875 =350 times 10minus3 and 492 versus 406 OR = 136 119875 = 002Supplementary Table S4) For SNP rs10975003 the frequencyof the heterozygous CT genotype was significantly higherin female AAU patients compared to the healthy controls(404 versus 336 OR = 157 119875 = 001 SupplementaryTable S5) A similar result was observed when we combinedCT and CC to construct a dominant model (443 versus380 OR = 157 119875 = 910times 10minus3 Supplementary Table S5)However none of the observed associations for the two SNPsretained statistical significance after Bonferroni correction(119875Bonferroni gt 005) Furthermore no significance associationswere found between the other SNPs and AAU even afterstratification by gender AS and HLA-B27 status (data notshown)
4 Discussion
In this study we first investigated whether genetic poly-morphisms of JAK-STAT signaling pathway genes includingJAK1 JAK2 STAT1 IRF1 and NOS2 confer susceptibilityto AAU with or without AS in a Chinese Han populationOur results suggest that none of these SNPs exhibit statisti-cally different frequencies of genotypes and alleles betweenhealthy controls and AAU patients However we observeda boundary significant association for two SNPs (rs10975003and rs10758669) of JAK2 by stratification analysis by genderand HLA-B27 status
We highlighted two issues at the time of study designto obtain unbiased association results First we followedstrict criteria for the diagnosis of AAU patients Patients withAAU were diagnosed as previously described by Jabs et al[36] and patients with AS were diagnosed with the modifiedNew York Criteria [37] In addition the AAU patients andhealthy controls were strictly matched by ethnicity and ageto avoid a possible influence of population stratificationFurthermore we only enrolled controls who had detailedhistories and physical examinations and excluded controlswith any autoimmune or immune-related diseases Finally
20 of the samples were randomly chosen and analyzed bydirect sequencing and the results of different genotypingmethods were consistent
AAU is defined as inflammation confined to the anteriorsegment of the eye that involves the iris and anterior part ofthe ciliary body The HLA-B27 is considered to be stronglyassociated with both AAU and AS [5 8 10 14] Recentgenetic studies have revealed that five candidate genes inthe JAK-STAT signaling pathway were considered geneticpredisposing factors for different autoimmune-mediated dis-eases [38ndash45] SNPs (rs10758669 and 10975003) in JAK2 areconsidered susceptibility factors for Crohnrsquos disease (CD) inthe German population and ulcerative colitis (UC) in theKorean population [38 45] One SNP (rs2070721) in IRF1 andSNPs (rs2297518 and rs4795067) in NOS2 are also associatedwith autoimmune diseases such as multiple sclerosis (MS)in Italy AS in Europe and psoriasis in Pakistan [39 43 44]Four SNPs (rs6718902 rs10199181 rs2066802 and rs1547550)in the STAT1 gene were observed to be associated with MSin Italy and IgA nephropathy (IgAN) in Korea [39 40] Inaddition an association was found between SNPs (rs2780815rs310241 rs3790532 rs310230 and rs310236) in JAK1 andBehcet disease (BD) as well as Vogt-Koyanagi-Harada (VKH)syndrome [41 42] two other common uveitis entities inChina There has been no report of associations betweenJAK-STAT signaling pathway genes and AAU and thus weperformed this case-control study to detect whether the fivecandidate genes were associated with AAU in a Chinese Hanpopulation Our results showed that there were no significantassociations between the genetic polymorphisms of the fivecandidate genes in the JAK-STAT signaling pathway andAAUThese results are not consistent with those observed forother autoimmune diseases reported in German Europeanand some other Asian populations [38 43 45] This discrep-ancy may be attributable to differences in the etiology andpathogenesis of AAU compared with BD VKH syndromeand autoimmune-mediated diseases
Consistent with our results a recent study also reportedno significant association of JAK-STAT signaling pathwaygene polymorphisms with rheumatoid arthritis stratified bythe presenceabsence of cardiovascular disease [46] Con-versely an influence of NFKB1 signaling pathway poly-morphisms on the development of cardiovascular events inpatients with rheumatoid arthritis has been observed [47]Furthermore NFKB1 signaling pathway polymorphismshave been described to play a critical role in the developmentof many autoimmune and inflammatory diseases and thusthe evaluation of the potential relationships between NFKB1polymorphisms and the development of AAU could be apromising research line for the future
There were several limitations of our studyWe had a lim-ited sample size to detect SNPs with weak effects while con-sidering multiple corrections Even for SNPs rs10758669 andrs10975003 we only had 700 power using a genetic powercalculator [48] In addition our samples were restricted to theHan Chinese population and all patients were enrolled fromthe ophthalmology department Further studies with a largersample size and other ethnic populations as well as patientsenrolled from multiple sources are warranted to confirm our
8 BioMed Research International
findings Additionally we only focused on eleven SNPs in theJAK-STAT pathway and it is possible that other unknownSNPs might be associated with AAU risk
In conclusion this study reveals that genetic polymor-phisms of JAK-STAT pathway genes including JAK1 JAK2STAT1 IRF1 andNOS2 may not be involved in susceptibilityto AAU risk in the Han Chinese population
Competing Interests
The authors declare that there is no conflict of interestsregarding the publication of this paper
Authorsrsquo Contributions
Ling Cheng and Hongsong Yu contributed equally to thiswork
Acknowledgments
The authors would like to thank Professor Peizeng Yang forhis great help in our study They are also grateful to alldonors enrolled in this study This work was supported bythe National Natural Science Foundation Project (8120067881670844) Fundamental and Advanced Research Programof Chongqing (cstc2015jcyjA10022) Science and TechnologyProject of Chongqing Municipal Education Commission(KJ1500236) Scientific Research Program of Science andTechnology Commission of Yuzhong District of Chongqing(20150102) and National Key Clinical Specialties Construc-tion Program of China
References
[1] D-K Hwang Y-J Chou C-Y Pu and P Chou ldquoEpidemi-ology of uveitis among the Chinese population in Taiwan APopulation-Based Studyrdquo Ophthalmology vol 119 no 11 pp2371ndash2376 2012
[2] K Nakao and N Ohba ldquoPrevalence of endogenous uveitis inKagoshima Prefecture Southwest JapanrdquoNipponGankaGakkaiZasshi vol 100 no 2 pp 150ndash155 1996
[3] D C Gritz and I GWong ldquoIncidence and prevalence of uveitisin Northern California the Northern California Epidemiologyof Uveitis Studyrdquo Ophthalmology vol 111 no 3 pp 491ndash5002004
[4] R B Nussenblatt and I Gery ldquoExperimental autoimmuneuveitis and its relationship to clinical ocular inflammatorydiseaserdquo Journal of Autoimmunity vol 9 no 5 pp 575ndash5851996
[5] P Yang Z Zhang H Zhou et al ldquoClinical patterns andcharacteristics of uveitis in a tertiary center for uveitis in ChinardquoCurrent Eye Research vol 30 no 11 pp 943ndash948 2005
[6] J T Gran and J F Skomsvoll ldquoThe outcome of ankylosingspondylitis a study of 100 patientsrdquo British Journal of Rheuma-tology vol 36 no 7 pp 766ndash771 1997
[7] L P Robertson andM J Davis ldquoA longitudinal study of diseaseactivity and functional status in a hospital cohort of patientswith ankylosing spondylitisrdquo Rheumatology vol 43 no 12 pp1565ndash1568 2004
[8] J H Chang P J McCluskey and D Wakefield ldquoAcute anterioruveitis and HLA-B27rdquo Survey of Ophthalmology vol 50 no 4pp 364ndash388 2005
[9] M Huhtinen and A Karma ldquoHLA-B27 typing in the categori-sation of uveitis in a HLA-B27 rich populationrdquo British Journalof Ophthalmology vol 84 no 4 pp 413ndash416 2000
[10] S Torres S Borges and A Artiles ldquoHLA-B27 and clinicalfeatures of acute anterior uveitis in Cubardquo Ocular Immunologyand Inflammation vol 21 no 2 pp 119ndash123 2013
[11] R Saari R Lahti K M Saari et al ldquoFrequency of rheumaticdiseases in patients with acute anterior uveitisrdquo ScandinavianJournal of Rheumatology vol 11 no 2 pp 121ndash123 1982
[12] A B Beckingsale J Davies J M Gibson and A RalphRosenthal ldquoAcute anterior uveitis ankylosing spondylitis backpain and HLA-B27rdquo British Journal of Ophthalmology vol 68no 10 pp 741ndash745 1984
[13] G P Thomas and M A Brown ldquoGenetics and genomics ofankylosing spondylitisrdquo Immunological Reviews vol 233 no 1pp 162ndash180 2010
[14] P C Robinson and M A Brown ldquoGenetics of ankylosingspondylitisrdquoMolecular Immunology vol 57 no 1 pp 2ndash11 2014
[15] C Lopez-Larrea K Sujirachato N K Mehra et al ldquoHLA-B27 subtypes in Asian patients with ankylosing spondylitisEvidence for new associationsrdquo Tissue Antigens vol 45 no 3pp 169ndash176 1995
[16] P C Robinson T A M Claushuis A Cortes et al ldquoGeneticdissection of acute anterior uveitis reveals similarities and dif-ferences in associations observed with ankylosing spondylitisrdquoArthritis and Rheumatology vol 67 no 1 pp 140ndash151 2015
[17] H Li S Hou H Yu et al ldquoAssociation of genetic variationsin TNFSF15 with acute anterior uveitis in Chinese HanrdquoInvestigative Ophthalmology and Visual Science vol 56 no 8pp 4605ndash4610 2015
[18] Q Xiang L Chen J Fang et al ldquoTNF receptor-associated factor5 gene confers genetic predisposition to acute anterior uveitisand pediatric uveitisrdquo Arthritis Research and Therapy vol 15no 5 article R113 2013
[19] H Yu Y Liu L Zhang et al ldquoFoxO1 gene confers genetic predis-position to acute anterior uveitis with ankylosing spondylitisrdquoInvestigative Ophthalmology amp Visual Science vol 55 no 12 pp7970ndash7974 2014
[20] T M Martin L Bye N Modi et al ldquoGenotype analysis ofpolymorphisms in autoimmune susceptibility genes CTLA-4and PTPN22 in an Acute anterior uveitis cohortrdquo MolecularVision vol 15 pp 208ndash212 2009
[21] Q Zhang J Qi S Hou et al ldquoA functional variant of PTPN22confers risk for Vogt-Koyanagi-Harada syndrome but not forankylosing spondylitisrdquo PLoS ONE vol 9 no 5 article e969432014
[22] S-S Zhao J-WHu JWang X-J Lou and L-L Zhou ldquoInversecorrelation between CD4+CD25 highCD127lowminus regulatory T-cells and serum immunoglobulin A in patients with new-onset ankylosing spondylitisrdquo Journal of International MedicalResearch vol 39 no 5 pp 1968ndash1974 2011
[23] W Zou Z Wu X Xiang S Sun and J Zhang ldquoThe expressionand significance of T helper cell subsets and regulatory Tcells CD4+CD25+ in peripheral blood of patients with humanleukocyte antigen B27-positive acute anterior uveitisrdquo GraefersquosArchive for Clinical and Experimental Ophthalmology vol 252no 4 pp 665ndash672 2014
BioMed Research International 9
[24] C Wang Q Liao Y Hu and D Zhong ldquoT lymphocyte subsetimbalances in patients contribute to ankylosing spondylitisrdquoExperimental and Therapeutic Medicine vol 9 no 1 pp 250ndash256 2015
[25] A Tedgui and Z Mallat ldquoCytokines in atherosclerosispathogenic and regulatory pathwaysrdquoPhysiological Reviews vol86 no 2 pp 515ndash581 2006
[26] P J Murray ldquoThe JAK-STAT signaling pathway Input andoutput integrationrdquo Journal of Immunology vol 178 no 5 pp2623ndash2629 2007
[27] A N Mathur H-C Chang D G Zisoulis et al ldquoStat3 andStat4 direct development of IL-17-secreting Th cellsrdquo Journal ofImmunology vol 178 no 8 pp 4901ndash4907 2007
[28] G K Hansson ldquoRegulation of immune mechanisms inatherosclerosisrdquo Annals of the New York Academy of Sciencesvol 947 pp 157ndash165 2001
[29] T Shea-Donohue A Fasano A Smith and A Zhao ldquoEntericpathogens and gut function role of cytokines and STATsrdquo GutMicrobes vol 1 no 5 pp 316ndash324 2010
[30] T Taniguchi K Ogasawara A Takaoka and N Tanaka ldquoIRFfamily of transcription factors as regulators of host defenserdquoAnnual Review of Immunology vol 19 pp 623ndash655 2001
[31] M Lohoff D Ferrick H-W Mittrucker et al ldquoInterferonregulatory factor-1 is required for a T helper 1 immune responsein vivordquo Immunity vol 6 no 6 pp 681ndash689 1997
[32] S Taki T Sato K Ogasawara et al ldquoMultistage regulation ofTh1-type immune responses by the transcription factor IRF-1rdquoImmunity vol 6 no 6 pp 673ndash679 1997
[33] C Bogdan ldquoNitric oxide and the immune responserdquo NatureImmunology vol 2 no 10 pp 907ndash916 2001
[34] S Brahmachari and K Pahan ldquoMyelin basic protein primingreduces the expression of Foxp3 in T cells via nitric oxiderdquoTheJournal of Immunology vol 184 no 4 pp 1799ndash1809 2010
[35] S-W Lee H Choi S-Y Eun S Fukuyama and M CroftldquoNitric oxide modulates TGF-120573-directive signals to suppressFoxp3 + regulatory T cell differentiation and potentiate Th1developmentrdquo Journal of Immunology vol 186 no 12 pp 6972ndash6980 2011
[36] D A Jabs R B Nussenblatt J T Rosenbaum and Standard-ization of Uveitis Nomenclature Working G ldquoStandardizationof uveitis nomenclature for reporting clinical data Resultsof the First International Workshoprdquo American Journal ofOphthalmology vol 140 pp 509ndash516 2005
[37] S Van Der Linden H A Valkenburg and A Cats ldquoEvaluationof diagnostic criteria for ankylosing spondylitis A proposal formodification of the New York criteriardquo Arthritis and Rheuma-tism vol 27 no 4 pp 361ndash368 1984
[38] D Ellinghaus E Ellinghaus R P Nair et al ldquoCombined anal-ysis of genome-wide association studies for Crohn disease andpsoriasis identifies seven shared susceptibility locirdquo AmericanJournal of Human Genetics vol 90 no 4 pp 636ndash647 2012
[39] G Fortunato G Calcagno V Bresciamorra et al ldquoMultiplesclerosis and hepatitis C virus infection are associated withsingle nucleotide polymorphisms in interferon pathway genesrdquoJournal of Interferon and Cytokine Research vol 28 no 3 pp141ndash152 2008
[40] W-H Hahn J-S Suh S H Cho B-S Cho and S-D KimldquoPolymorphisms of signal transducers and activators of tran-scription 1 and 4 (STAT1 and STAT4) contribute to progressionof childhood IgA nephropathyrdquo Cytokine vol 50 no 1 pp 69ndash74 2010
[41] S Hou J Qi Q Zhang et al ldquoGenetic variants in the JAK1 geneconfer higher risk of Behcetrsquos disease with ocular involvementin Han ChineserdquoHumanGenetics vol 132 no 9 pp 1049ndash10582013
[42] K Hu S Hou F Li Q Xiang A Kijlstra and P YangldquoJAK1 but not JAK2 and STAT3 confers susceptibility toVogt-Koyanagi-Harada (VKH) syndrome in a Han Chinesepopulationrdquo Investigative Ophthalmology and Visual Sciencevol 54 no 5 pp 3360ndash3365 2013
[43] A Cortes J Hadler J P Pointon et al ldquoIdentification ofmultiple risk variants for ankylosing spondylitis through high-density genotyping of immune-related locirdquo Nature Geneticsvol 45 no 7 pp 730ndash738 2013
[44] P A Shaiq P E Stuart A Latif et al ldquoGenetic associations ofpsoriasis in a Pakistani populationrdquo British Journal of Derma-tology vol 169 no 2 pp 406ndash411 2013
[45] S-K Yang Y Jung H Kim M Hong B D Ye and KSong ldquoAssociation of FCGR2A JAK2 or HNF4A variants withulcerative colitis in Koreansrdquo Digestive and Liver Disease vol43 no 11 pp 856ndash861 2011
[46] M Garcıa-Bermudez R Lopez-Mejıas F Genre et al ldquoLackof association between JAK3 gene polymorphisms and cardio-vascular disease in Spanish patients with rheumatoid arthritisrdquoBioMed Research International vol 2015 Article ID 318364 11pages 2015
[47] R Lopez-Mejıas M Garcıa-Bermudez C Gonzalez-Juanateyet al ldquoNFKB1-94ATTG insdel polymorphism (rs28362491) isassociated with cardiovascular disease in patients with rheuma-toid arthritisrdquo Atherosclerosis vol 224 no 2 pp 426ndash429 2012
[48] H J Park J W Kim B-S Cho and J-H Chung ldquoAssociationof BH3 interacting domain death agonist (BID) gene polymor-phisms with proteinuria of immunoglobulin A nephropathyrdquoScandinavian Journal of Clinical and Laboratory Investigationvol 74 no 4 pp 329ndash335 2014
Submit your manuscripts athttpwwwhindawicom
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
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PeptidesInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporation httpwwwhindawicom
International Journal of
Volume 2014
Zoology
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Molecular Biology International
GenomicsInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
BioinformaticsAdvances in
Marine BiologyJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Signal TransductionJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
BioMed Research International
Evolutionary BiologyInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
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Biochemistry Research International
ArchaeaHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Genetics Research International
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Advances in
Virolog y
Hindawi Publishing Corporationhttpwwwhindawicom
Nucleic AcidsJournal of
Volume 2014
Stem CellsInternational
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
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Enzyme Research
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
International Journal of
Microbiology
2 BioMed Research International
an important role in immune responses [29] IRF1 is the firstmember identified in the IRF family and is involved in manyinnate and adaptive immune responses [30] Impaired orabsent Th1-type immune responses favor Th2 differentiationin IRF1-deficientmice [31 32]NOS2-derivedNO a key factorin immunoregulation [33] can inhibit Th1 as well as Th2cytokine production and regulate the development of FoxP3+Treg cells [34 35] In summary JAK-STAT signaling pathwaygenes including JAK1 JAK2 STAT1 IRF1 and NOS2 havebeen suggested to be strongly linked with T cells and may beinvolved in the pathophysiology of AAU with or without AS
Thus we conducted the present case-control study toinvestigate whether JAK-STAT signaling pathway genes con-fer susceptibility to AAU risk in a Chinese Han population
2 Materials and Methods
21 Subjects A total of 929 AAU patients were enrolled inthis study including 443 patients with AS (AAU+AS+) and486 patients without AS (AAU+ASminus) as well as 714 gender-and race-matched healthy controls All subjects were HanChinese recruited from the Department of Ophthalmologyin the First Affiliated Hospital of Chongqing Medical Uni-versity (Chongqing China) between June 2008 and May2015 All AAU patients were diagnosed based on medicalrecords physical examinations and the anatomic locationof inflammation as previously described by Jabs et al [36]The diagnosis of AS followed the modified New York Criteria[37] All subjects gave written informed consent before bloodcollection This study was approved by the Human EthicsCommittee of the First Affiliated Hospital of ChongqingMedical University (Approval number 2009-201008) andfollowed the tenets of the Declaration of Helsinki
22 SNP Selection We selected candidate single nucleotidepolymorphisms (SNPs) based on previously published stud-ies and included only those SNPs significantly associatedwith autoimmune diseases [38ndash45] We used HaploView 42software to evaluate the linkage disequilibrium (LD) andminor allele frequency (MAF) of the SNPs Five SNPs ofJAK1 rs2780815 rs3790532 rs310230 rs310236 and rs310241[41 42] were selected Since the SNPs rs3790532 rs310230rs310236 and rs310241 are in strong LD with each other (1199032 gt08 Figure 1) we only used rs310241 in our study Further-more we also eliminated SNPs that were not polymorphicin the Chinese population Finally eleven SNPs in five JAK-STAT signaling pathway genes were tested in our studyincluding two SNPs in the intron region of the JAK1 gene(rs310241 rs2780815) [41] two SNPs in the exon region and31015840UTR of the JAK2 gene (rs10758669 rs10975003) [38 45]one SNP in the intron region of the IRF1 gene (rs2070721)four SNPs in the exon region and intron region of the STAT1gene (rs2066802 rs1547550 rs6718902 and rs10199181) [3940] and two SNPs in the exon region and intron region ofthe NOS2 gene (rs2297518 rs4795067) [43 44]
23 DNA Extraction and Genotyping Peripheral blood sam-ples were collected from subjects and genomic DNA extrac-tion was performed using the QIAamp DNA Blood Mini
84
84 83
83
11
11
10
3 11 12 71 83
rs2780815
rs310241
rs3790532
rs310236
rs310230
10
Block 1 (37kb)
Figure 1 Linkage disequilibrium (LD) analysis of rs2780815rs310241 rs3790532 rs310236 and rs310230 in the JAK1 gene TheLD block was estimated by HaploView software version 42 usingthe Chinese Han HapMap data The number in the square indicatesthe 1199032 value
Kit (Qiagen Valencia CA USA) The genomic DNA wasquantified with NanoDrop 2000 (Thermal Fisher Scien-tific Delaware USA) and stored at minus20∘C until use ThreeSNPs (rs2780815 rs2070721 and rs10199181) were genotypedby TaqMan probe (Applied Biosystems Foster City CA)and the others were genotyped by polymerase chain reac-tion restriction fragment length polymorphism (PCR-RFLP)assay The specific primers for PCR and restriction enzymesare described in Table 1 The PCR reactions were performedunder the following conditions denaturation at 95∘C for5 minutes 33 to 36 cycles of denaturation at 95∘C for 30seconds annealing at 56ndash64∘C for 30 s and extension at 72∘Cfor 30 seconds and a final extension at 72∘C for 5 minutesThe enzyme-digested products were visualized on 3 or4 agarose gels and stained with GoldView (SBS GenetechBeijing China) Direct sequencing was carried out randomlyon 10 of the study samples to assure the validity of theSNP genotyping method used The success rate of all SNPgenotyping ranged from 973 to 100
24 StatisticalAnalysis Hardy-Weinberg equilibrium (HWE)was analyzed by the 1205942 test The distributions of the alleleand genotype frequencies between the patients and controlswere compared by the 1205942 test Multivariate logistic regressionmodel adjusted for age and gender was further adopted totest the associations between the SNPs and AAU The riskeffect of each SNP was measured by odds ratios (OR) and95confidence intervals (CI)Therewere four differentmod-els of inheritance in our study including additive modelscodominantmodels dominantmodels and recessivemodels119875 values were corrected using the Bonferroni correction
BioMed Research International 3
Table 1 PCR primer sequences and restriction enzymes
SNP Primers Restriction enzyme
rs310241 51015840 AACCACCAGCTCAACATTCCTAG 3101584051015840 CAGCCAGGTCTCCCGTAGG 31015840 BseDI
rs6718902 51015840 CGGACAAAAGCATGCACTAGA 3101584051015840 CCACCACCATTAATAGGTGACTTTA 31015840 DraI
rs2297518 51015840 TGAGCTCTTTCAGCATGAAGATC 3101584051015840 CTTCCGTGGTGGGCTGTG 31015840 TaqI
rs10758669 51015840 TGATGTAGAGACAAGGACATGCTGAGGTAC 3101584051015840 GCCAAAAGACAAAGGCAAGGGG 31015840 BanI
rs10975003 51015840 GGCCAGTCAAGAAAAAACCAGTT 3101584051015840 TTCGGAGTCTTGTCTGAGCATGT 31015840 HpaI
rs4795067 51015840 GCACTCATTCATTCATGCAAACATA 3101584051015840 GGCAGAACTTGAACCCCAGCT 31015840 NdeI
rs1547550 51015840 CTTCTCTAGGAGGCCCAGCA 3101584051015840 TGGGCACCACGATATGAGAG 31015840 BstMAI
rs2066802 51015840 ATTCCTGGAGCAGGTTCACAAG 3101584051015840 AAACATGGCCCCAAGTCACT 31015840 HindIII
method considering multiple tests Statistical significancelevel was defined as a corrected 119875 value lt 005 Statisticalanalyses were performed using SPSS version 170 (SPSS IncChicago IL USA)
3 Results
31 Clinical Features of AAU Patients The detailed clinicalfeatures and demographic characteristics of the AAUpatientsare presented in Table 2 All 929 AAU patients include 569(612) males and 360 (388) females The 714 controlsubjects consisted of 428 (599) males and 286 (401)females The average age was 398 plusmn 123 in AAU patientsand 395 plusmn 108 in the controls respectively There were nosignificant differences in age and gender between the casesand controls In addition 546 AAU patients (689) wereHLA-B27-positive whereas 246 AAU patients (311) wereHLA-B27-negative
32 The Genotype and Allele Frequency Distribution of theTested SNPs in AAU Eleven SNPs of the JAK-STAT signalingpathway genes (JAK1 JAK2 STAT1 IRF1 and NOS2) weresuccessfully genotyped There were no significant deviationsof HWE in either the cases or controls We did not observesignificant differences in the genotype and allele distribu-tions of any of the SNPs between the AAU patients andcontrol subjects after Bonferroni correction (see Supplemen-tary Table S1 in Supplementary Material available onlineat httpdxdoiorg10115520165896906) Further stratifiedanalyses of gender AS andHLA-B27 status showed a bound-ary significant association of two SNPs (rs10975003 andrs10758669) of JAK2 with AAU In female AS-positive AAUpatients there was a decreased frequency of the TT genotypeof rs10975003 compared to the female controls (OR = 055119875 = 159 times 10minus3 119875Bonferroni = 0052 Table 3) whereas nosignificant differences were observed in the genotype andallele frequencies of the other ten SNPs (119875Bonferroni gt 005Table 3) Similarly there were no significant differences in
Table 2 Clinical characteristics of the investigated subjects
Clinical features Total PercentageAAU patients 929 100Mean age plusmn SD (years) 398 plusmn 123AAU with AS 443 477AAU without AS 486 523AAU male 569 612AAU female 360 388AAU with AS (male) 326 (443 tested) 736AAU with AS (female) 117 (443 tested) 264AAU without AS (male) 243 (486 tested) 50AAU without AS (female) 243 (486 tested) 50HLA-B27+ AAU 546 (792 tested) 689HLA-B27+ AAU AS+ 348 (428 tested) 813HLA-B27+ AAU ASminus 198 (364 tested) 544Control 714 100Mean age plusmn SD (years) 395 plusmn 108Male 428 599Female 286 401
the genotype and allele frequencies of the SNPs between themale AAU patients and male controls (119875Bonferroni gt 005Supplementary Table S2)
In addition an increased frequency of theAC genotype inrs10758669 was observed in HLA-B27-positive AAU patientscompared to the healthy controls (OR = 144119875 = 162times10minus3119875Bonferroni = 0053 Table 4) whereas there were no significantdifferences in the genotype and allele frequencies of the otherten SNPs between the HLA-B27-positive AAU patients andthe control subjects (119875Bonferroni gt 005 Table 4)
However an increased frequency of the rs10758669ACgenotype was observed in HLA-B27-positive AS-positiveAAU patients compared to healthy controls (OR = 149 119875 =256times10minus3119875Bonferroni = 0084 Table 5) whereas no significant
4 BioMed Research InternationalTa
ble3Alleleandgeno
type
frequ
encies
infemaleA
AUpatie
ntsa
ndfemalec
ontro
ls
Gene
SNP
Alleleandgeno
type
AAU+AS+
(female)
AAU+ASminus
(female)
Con
trols(fe
male)119875(A
S+)
Pc(A
S+)
OR(95
CI)119875(A
Sminus)
Pc(A
Sminus)
OR(95
CI)
JAK1
rs310241
C80
(333
)138(296
)
156(287
)019
NS
124(090ndash
172)
074
NS
105(080ndash
137)
CC12
(100
)14
(60)
17(62)
019
NS
167(077ndash361)
091
NS
096
(046ndash
200)
CT56
(467
)110(472
)
122(449
)074
NS
108(070ndash
166)
060
NS
110(077ndash15
6)TT
52(433
)109(468
)133(489
)031
NS
080
(052
ndash123)
064
NS
092
(065ndash13
0)
rs2780815
G159(787
)40
0(870
)
495(853
)003
NS
064
(042ndash096)
045
NS
115(080ndash
163)
GG
63(624
)175(761
)212(731
)004
NS
061
(038ndash
099)
044
NS
117(079ndash
175)
GT
33(326
)50
(217
)
71(245
)011
NS
150(091ndash245)
046
NS
086
(057ndash12
9)TT
5(50)
5(22)
7(24)
020
NS
211(065ndash680)
086
NS
090
(028ndash287)
JAK2
rs10758669
A170(739
)328(675
)
399(723
)064
NS
109(077ndash15
4)010
NS
080
(061ndash10
4)AA
59(513
)
104(428
)144(522
)088
NS
097
(063ndash14
9)003
NS
069
(049ndash
100)
AC52
(452
)120(494
)
111(40
2)
036
NS
123(079ndash
190)
004
NS
145(102ndash205)
CC4(35)
19(78
)21
(76
)013
NS
044
(015
ndash130)
093
NS
103(054ndash
197)
rs10975003
C44
(206
)117
(258
)93
(179
)
040
NS
119(080ndash
177)
283times10minus3
009
159(117ndash217
)CC
2(19)
11(48)
8(31
)052
NS
060
(013
ndash287)
032
NS
160(063ndash406)
CT40
(374
)
95(419
)
77(296
)
015
NS
142(088ndash228)
483times10minus3
NS
171(118ndash249)
TT65
(607
)121(533)
175(673
)
023
NS
075
(047ndash12
0)15
9times10minus3
0052
055
(038ndash
080)
STAT
1
rs1547550
C205(869
)411(886)
423(870
)
095
NS
099
(062ndash15
6)047
NS
116(078ndash17
1)CC
88(746
)182(784
)185(761
)075
NS
092
(055ndash15
3)055
NS
114(074
ndash175)
CG29
(246
)47
(203
)53
(218
)
056
NS
117(070ndash
196)
068
NS
091
(059ndash
142)
GG
1(08
)3(13)
5(21
)040
NS
041
(005ndash352)
056
NS
065
(015
ndash276
)
rs2066
802
C46
(204
)81
(174
)
113(212
)
080
NS
095
(065ndash14
0)013
NS
078
(057ndash10
8)CC
1(09
)6(26)
8(30)
022
NS
029
(004ndash
234)
078
NS
086
(029ndash
250)
CT44
(389
)69
(296
)
97(363
)063
NS
112(071ndash17
6)011
NS
074
(051ndash10
7)TT
68(602
)158(678
)
162(607
)093
NS
098
(063ndash15
4)010
NS
137(095ndash19
7)
rs6718902
C132(600
)259(542
)308(538
)012
NS
129(094ndash
176)
091
NS
101(079ndash129)
CC36
(327
)72
(301
)82
(287
)043
NS
121(075ndash194)
072
NS
107(074
ndash156)
CT60
(545
)115(481
)144(503
)045
NS
118(076
ndash184)
061
NS
092
(065ndash12
9)TT
14(127
)52
(218
)
60(210
)
006
NS
055
(029ndash
103)
083
NS
105(069ndash
159)
rs10199181
A51
(248
)117
(261
)148(252
)091
NS
098
(068ndash14
1)073
NS
105(079ndash
139)
AA
6(58)
20(89)
19(65)
082
NS
090
(035ndash231)
029
NS
142(074
ndash273)
AT39
(379
)
77(344
)110(374
)
094
NS
102(064ndash
162)
048
NS
088
(061ndash12
6)TT
58(563
)127(567
)165(561
)097
NS
101(064
ndash158)
090
NS
102(072ndash14
5)
IRF1
rs2070721
A86
(406
)154(339
)216(343
)010
NS
131(095ndash180)
090
NS
098
(076
ndash127)
AA
15(14
2)
26(115
)
29(92)
015
NS
163(084ndash
317)
039
NS
128(073ndash223)
AC56
(528
)102(449
)158(502
)063
NS
111(072ndash173)
023
NS
081
(058ndash114)
CC35
(330
)99
(436
)128(406
)016
NS
072
(045ndash114)
049
NS
113(080ndash
160)
NOS2
rs2297518
A52
(205
)69
(152
)74
(157
)010
NS
139(094ndash
205)
084
NS
096
(068ndash13
8)AA
3(24)
6(26)
2(08)
024
NS
283
(047ndash1717)
014
NS
318
(063ndash1590)
AG46
(362
)57
(252
)70
(297
)
020
NS
135(085ndash213)
027
NS
080
(053
ndash120)
GG
78(614
)
164(722
)164(695
)
012
NS
070
(045ndash110)
051
NS
114(077ndash17
1)
rs4795067
A173(739
)369(759
)439(778
)
024
NS
081
(057ndash115)
046
NS
090
(067ndash12
0)AA
62(530
)145(597
)
169(599
)
020
NS
075
(049ndash
116)
095
NS
099
(070ndash
140)
AG49
(419
)
79(325
)101(358)
026
NS
129(083ndash201)
043
NS
086
(060ndash
124)
GG
6(51
)19
(78
)12
(43)
070
NS
122(045ndash332)
008
NS
191(091ndash4
02)
OR=od
dsratio
95
CI=95
confi
denceinterval
Pc=119875valuea
djustedby
Bonferroni
correctio
n
BioMed Research International 5
Table 4 Allele and genotype frequencies of SNPs in patients with AAU versus control subjects stratified by HLA-B27 status
Gene SNP Allele and genotype AAUHLA-B27 Control 119875 Pc OR (95 CI)
JAK1
rs310241
C 324 (310) 392 (275) 006 NS 118 (100ndash141)CC 41 (79) 45 (63) 029 NS 127 (082ndash196)CT 242 (464) 302 (424) 016 NS 118 (094ndash148)TT 239 (457) 366 (513) 005 NS 080 (064ndash100)
rs2780815
G 878 (862) 1250 (875) 035 NS 089 (070ndash113)GG 381 (748) 548 (767) 044 NS 090 (069ndash118)GT 116 (228) 154 (216) 061 NS 107 (082ndash141)TT 12 (24) 12 (17) 040 NS 141 (063ndash317)
JAK2
rs10758669
A 705 (659) 980 (690) 010 NS 087 (073ndash103)AA 220 (411) 346 (487) 008 NS 074 (059ndash092)AC 265 (495) 288 (406) 162 times 10minus3 0053 144 (115ndash180)CC 50 (94) 76 (107) 043 NS 086 (059ndash125)
rs10975003
C 251 (245) 272 (212) 005 NS 121 (100ndash147)CC 26 (51) 28 (44) 056 NS 118 (068ndash203)CT 199 (388) 216 (335) 006 NS 126 (099ndash160)TT 287 (561) 399 (621) 003 NS 077 (061ndash098)
STAT1
rs1547550
C 937 (872) 1254 (878) 067 NS 095 (075ndash121)CC 407 (758) 555 (777) 042 NS 090 (069ndash117)CG 123 (229) 144 (202) 024 NS 118 (090ndash154)GG 7 (13) 15 (21) 029 NS 062 (025ndash152)
rs2066802
C 190 (193) 288 (207) 039 NS 091 (074ndash112)CC 15 (30) 27 (39) 044 NS 078 (041ndash148)CT 160 (325) 234 (337) 066 NS 095 (074ndash121)TT 318 (645) 434 (624) 047 NS 109 (086ndash139)
rs6718902
C 584 (565) 778 (545) 033 NS 108 (092ndash127)CC 163 (315) 209 (293) 040 NS 111 (087ndash142)CT 258 (499) 360 (504) 086 NS 098 (078ndash123)TT 96 (186) 145 (203) 045 NS 090 (067ndash119)
rs10199181
A 270 (272) 397 (278) 075 NS 097 (081ndash116)AA 43 (87) 55 (77) 055 NS 114 (075ndash173)AT 184 (371) 287 (402) 028 NS 088 (069ndash111)TT 269 (542) 372 (521) 047 NS 109 (087ndash137)
IRF1 rs2070721
A 356 (357) 487 (341) 040 NS 108(091ndash127)AA 61 (122) 74 (104) 030 NS 121 (084ndash173)AC 234 (470) 339 (474) 087 NS 098 (078ndash123)CC 203 (408) 301 (422) 063 NS 094 (075ndash119)
NOS2
rs2297518
A 194 (181) 209 (162) 021 NS 115 (093ndash142)AA 12 (23) 12 (19) 064 NS 121 (054ndash272)AG 170 (317) 185 (286) 024 NS 116 (090ndash149)GG 354 (660) 450 (695) 020 NS 085 (067ndash109)
rs4795067
A 797 (743) 1008 (761) 035 NS 092 (076ndash110)AA 293 (546) 382 (577) 033 NS 089 (071ndash123)AG 211 (394) 244 (369) 037 NS 111 (088ndash141)GG 32 (60) 36 (54) 069 NS 110 (068ndash180)
OR = odds ratio 95 CI = 95 confidence intervalPc = 119875 value adjusted by Bonferroni correction
6 BioMed Research InternationalTa
ble5Alleleandgeno
type
frequ
encies
ofSN
Psin
patientsw
ithAAU
versus
controlsub
jectsstratified
byASandHLA
-B27
status
Gene
SNP
Alleleandgeno
type
AAU+AS+
HLA
-B27
AAU+ASminus
HLA
-B27
Con
trol
119875(A
S+)
Pc(A
S+)
OR(95
CI)119875(A
Sminus)
Pc(A
Sminus)
OR(95
CI)
JAK1
rs310241
C205(316
)
119(299
)
392(275
)
005
NS
122(100ndash
149)
032
NS
113(088ndash14
5)CC
30(93)
11(55)
45(63)
009
NS
152(094ndash
245)
070
NS
087
(044ndash
172)
CT145(447
)97
(487
)302(424
)047
NS
110(085ndash14
4)010
NS
131(095ndash179)
TT149(460
)90
(457
)366(513
)
011
NS
081
(062ndash10
5)014
NS
079
(058ndash10
8)
rs2780815
G543(843
)335(896
)
1250
(875
)
005
NS
077
(059ndash
100)
028
NS
122(085ndash17
7)GG
230(714
)
151(807)
548(767
)007
NS
076
(056ndash
102)
024
NS
127(085ndash19
0)GT
83(258
)33
(176
)
154(216
)
014
NS
126(093ndash17
2)023
NS
078
(051ndash118)
TT9(28)
3(17)
12(17)
024
NS
168(070ndash
403)
094
NS
095
(027ndash342)
JAK2
rs10758669
A44
9(662
)256(653
)980(690
)
020
NS
088
(072ndash17
0)016
NS
085
(067ndash10
7)AA
139(410
)
81(413
)
346(487
)002
NS
073
(056ndash
095)
007
NS
074
(054ndash
102)
AC171(504)
94(480
)288(406
)256times10minus3
0084
149(115
ndash194)
006
NS
135(098ndash18
6)CC
29(86)
21(107
)76
(107
)028
NS
078
(050ndash
122)
100
NS
100(060ndash
167)
rs10975003
C165(257
)86
(225
)272(212
)
003
NS
129(103ndash16
1)057
NS
108(082ndash14
3)CC
18(56)
8(42)
28(44)
039
NS
131(071ndash240)
092
NS
096
(043ndash214)
CT129(402
)70
(366
)216(335
)004
NS
133(101ndash17
5)044
NS
114(082ndash16
0)TT
174(542
)113(592
)
399(621
)001
NS
072
(055ndash095)
047
NS
089
(064ndash
123)
STAT
1
rs1547550
C608(881
)329(857
)1254
(878
)
084
NS
103(078ndash13
6)026
NS
083
(060ndash
115)
CC264(765
)143(745
)555(777
)
066
NS
093
(069ndash
127)
034
NS
084
(058ndash12
1)CG
80(232
)43
(224
)144(202
)026
NS
120(088ndash16
3)050
NS
114(078ndash16
8)GG
1(03
)6(31
)15
(21
)002
NS
014
(002ndash103)
040
NS
150(058ndash393)
rs2066
802
C130(207
)60
(168
)288(207
)099
NS
100(079ndash
126)
009
NS
077
(057ndash10
5)CC
11(35)
4(22)
27(39)
077
NS
090
(044ndash
183)
029
NS
057
(020ndash
164)
CT108(344
)52
(291
)
234(337
)082
NS
103(078ndash13
7)024
NS
081
(056ndash
116)
TT195(621
)123(687
)434(624
)092
NS
099
(075ndash13
0)012
NS
132(093ndash18
8)
rs6718902
C368(577
)
216(545
)778(545
)018
NS
114(094ndash
138)
098
NS
100(080ndash
125)
CC103(323
)60
(303
)209(293
)
033
NS
115(087ndash15
3)078
NS
105(075ndash14
8)CT
162(508
)96
(485
)360(504
)091
NS
102(078ndash13
2)063
NS
093
(068ndash12
7)TT
54(169
)42
(212
)
145(203
)020
NS
080
(057ndash113)
078
NS
106(072ndash15
6)
rs10199181
A170(269
)100(278
)
397(278
)
067
NS
096
(077ndash118)
099
NS
100(077ndash12
9)AA
25(79
)18
(100
)55
(77
)091
NS
103(063ndash16
8)032
NS
133(076
ndash233
)AT
120(380
)64
(356
)287(402
)050
NS
091
(069ndash
120)
026
NS
082
(058ndash115)
TT171(541
)98
(544
)372(521
)055
NS
108(083ndash14
1)057
NS
110(079ndash
153)
IRF1
rs2070721
A221(349)
135(373
)
487(341
)073
NS
103(085ndash12
6)025
NS
115(090-14
6)AA
37(117
)
24(133
)74
(104
)053
NS
114(075ndash174)
026
NS
132(081ndash216)
AC147(464
)87
(481
)339(474
)
074
NS
096
(073ndash12
5)089
NS
102(074
ndash142)
CC133(420
)70
(386
)301(422)
095
NS
099
(076
ndash130)
040
NS
097
(062ndash12
1)
NOS2
rs2297518
A130(19
0)
64(165
)209(162
)011
NS
122(096ndash
155)
087
NS
103(076
ndash139
)AA
7(20)
5(26)
12(19)
083
NS
111(043ndash284)
053
NS
140(049ndash
402)
AG116(339
)54
(278
)
185(286
)008
NS
128(097ndash17
0)084
NS
096
(067ndash13
8)GG
219(641
)135(696
)
450(695
)
007
NS
078
(059ndash
103)
099
NS
100(071ndash14
2)
rs4795067
A512(736
)295(745
)1008
(761
)020
NS
087
(071ndash10
8)061
NS
092
(071ndash119)
AA
184(529
)109(551
)382(577
)
014
NS
082
(063ndash10
7)051
NS
090
(065ndash12
4)AG
144(414
)
77(388
)244(369
)016
NS
121(093ndash158)
060
NS
109(079ndash
151)
GG
20(57)
12(61
)36
(54)
084
NS
106(060ndash
186)
074
NS
112(057ndash220)
OR=od
dsratio
95
CI=95
confi
denceinterval
Pc=119875valuea
djustedby
Bonferroni
correctio
n
BioMed Research International 7
differences in the genotype and allele frequencies of the other10 SNPs were observed between the HLA-B27-positive AS-positiveAAUpatients and healthy controls (119875Bonferroni gt 005Table 5) In addition there were no significant differencesin the genotype and allele frequencies of the tested SNPsbetween the AS-positive AAU patients and control subjects(119875Bonferroni gt 005 Supplementary Table S3)
33 Logistic Regression Analysis of SNPs in AAU We furtherinvestigated the SNPs rs10758669 and rs10975003 using addi-tive codominant dominant and recessive genetic modelsusing a multivariate logistic regression model adjusted forage and gender We observed that the frequency of the CAgenotype of rs10758669 was significantly higher in AAUpatients which suggests that patients with the rs10758669CA genotype have increased susceptibility to AAU (468versus 406 OR = 128 119875 = 002 SupplementaryTable S4) An increased frequency of the AC genotype ofrs10758669 was also observed in HLA-B27-positive AAUpatients and AS-positive AAU patients compared to thehealthy controls (495 versus 406 OR = 143 119875 =350 times 10minus3 and 492 versus 406 OR = 136 119875 = 002Supplementary Table S4) For SNP rs10975003 the frequencyof the heterozygous CT genotype was significantly higherin female AAU patients compared to the healthy controls(404 versus 336 OR = 157 119875 = 001 SupplementaryTable S5) A similar result was observed when we combinedCT and CC to construct a dominant model (443 versus380 OR = 157 119875 = 910times 10minus3 Supplementary Table S5)However none of the observed associations for the two SNPsretained statistical significance after Bonferroni correction(119875Bonferroni gt 005) Furthermore no significance associationswere found between the other SNPs and AAU even afterstratification by gender AS and HLA-B27 status (data notshown)
4 Discussion
In this study we first investigated whether genetic poly-morphisms of JAK-STAT signaling pathway genes includingJAK1 JAK2 STAT1 IRF1 and NOS2 confer susceptibilityto AAU with or without AS in a Chinese Han populationOur results suggest that none of these SNPs exhibit statisti-cally different frequencies of genotypes and alleles betweenhealthy controls and AAU patients However we observeda boundary significant association for two SNPs (rs10975003and rs10758669) of JAK2 by stratification analysis by genderand HLA-B27 status
We highlighted two issues at the time of study designto obtain unbiased association results First we followedstrict criteria for the diagnosis of AAU patients Patients withAAU were diagnosed as previously described by Jabs et al[36] and patients with AS were diagnosed with the modifiedNew York Criteria [37] In addition the AAU patients andhealthy controls were strictly matched by ethnicity and ageto avoid a possible influence of population stratificationFurthermore we only enrolled controls who had detailedhistories and physical examinations and excluded controlswith any autoimmune or immune-related diseases Finally
20 of the samples were randomly chosen and analyzed bydirect sequencing and the results of different genotypingmethods were consistent
AAU is defined as inflammation confined to the anteriorsegment of the eye that involves the iris and anterior part ofthe ciliary body The HLA-B27 is considered to be stronglyassociated with both AAU and AS [5 8 10 14] Recentgenetic studies have revealed that five candidate genes inthe JAK-STAT signaling pathway were considered geneticpredisposing factors for different autoimmune-mediated dis-eases [38ndash45] SNPs (rs10758669 and 10975003) in JAK2 areconsidered susceptibility factors for Crohnrsquos disease (CD) inthe German population and ulcerative colitis (UC) in theKorean population [38 45] One SNP (rs2070721) in IRF1 andSNPs (rs2297518 and rs4795067) in NOS2 are also associatedwith autoimmune diseases such as multiple sclerosis (MS)in Italy AS in Europe and psoriasis in Pakistan [39 43 44]Four SNPs (rs6718902 rs10199181 rs2066802 and rs1547550)in the STAT1 gene were observed to be associated with MSin Italy and IgA nephropathy (IgAN) in Korea [39 40] Inaddition an association was found between SNPs (rs2780815rs310241 rs3790532 rs310230 and rs310236) in JAK1 andBehcet disease (BD) as well as Vogt-Koyanagi-Harada (VKH)syndrome [41 42] two other common uveitis entities inChina There has been no report of associations betweenJAK-STAT signaling pathway genes and AAU and thus weperformed this case-control study to detect whether the fivecandidate genes were associated with AAU in a Chinese Hanpopulation Our results showed that there were no significantassociations between the genetic polymorphisms of the fivecandidate genes in the JAK-STAT signaling pathway andAAUThese results are not consistent with those observed forother autoimmune diseases reported in German Europeanand some other Asian populations [38 43 45] This discrep-ancy may be attributable to differences in the etiology andpathogenesis of AAU compared with BD VKH syndromeand autoimmune-mediated diseases
Consistent with our results a recent study also reportedno significant association of JAK-STAT signaling pathwaygene polymorphisms with rheumatoid arthritis stratified bythe presenceabsence of cardiovascular disease [46] Con-versely an influence of NFKB1 signaling pathway poly-morphisms on the development of cardiovascular events inpatients with rheumatoid arthritis has been observed [47]Furthermore NFKB1 signaling pathway polymorphismshave been described to play a critical role in the developmentof many autoimmune and inflammatory diseases and thusthe evaluation of the potential relationships between NFKB1polymorphisms and the development of AAU could be apromising research line for the future
There were several limitations of our studyWe had a lim-ited sample size to detect SNPs with weak effects while con-sidering multiple corrections Even for SNPs rs10758669 andrs10975003 we only had 700 power using a genetic powercalculator [48] In addition our samples were restricted to theHan Chinese population and all patients were enrolled fromthe ophthalmology department Further studies with a largersample size and other ethnic populations as well as patientsenrolled from multiple sources are warranted to confirm our
8 BioMed Research International
findings Additionally we only focused on eleven SNPs in theJAK-STAT pathway and it is possible that other unknownSNPs might be associated with AAU risk
In conclusion this study reveals that genetic polymor-phisms of JAK-STAT pathway genes including JAK1 JAK2STAT1 IRF1 andNOS2 may not be involved in susceptibilityto AAU risk in the Han Chinese population
Competing Interests
The authors declare that there is no conflict of interestsregarding the publication of this paper
Authorsrsquo Contributions
Ling Cheng and Hongsong Yu contributed equally to thiswork
Acknowledgments
The authors would like to thank Professor Peizeng Yang forhis great help in our study They are also grateful to alldonors enrolled in this study This work was supported bythe National Natural Science Foundation Project (8120067881670844) Fundamental and Advanced Research Programof Chongqing (cstc2015jcyjA10022) Science and TechnologyProject of Chongqing Municipal Education Commission(KJ1500236) Scientific Research Program of Science andTechnology Commission of Yuzhong District of Chongqing(20150102) and National Key Clinical Specialties Construc-tion Program of China
References
[1] D-K Hwang Y-J Chou C-Y Pu and P Chou ldquoEpidemi-ology of uveitis among the Chinese population in Taiwan APopulation-Based Studyrdquo Ophthalmology vol 119 no 11 pp2371ndash2376 2012
[2] K Nakao and N Ohba ldquoPrevalence of endogenous uveitis inKagoshima Prefecture Southwest JapanrdquoNipponGankaGakkaiZasshi vol 100 no 2 pp 150ndash155 1996
[3] D C Gritz and I GWong ldquoIncidence and prevalence of uveitisin Northern California the Northern California Epidemiologyof Uveitis Studyrdquo Ophthalmology vol 111 no 3 pp 491ndash5002004
[4] R B Nussenblatt and I Gery ldquoExperimental autoimmuneuveitis and its relationship to clinical ocular inflammatorydiseaserdquo Journal of Autoimmunity vol 9 no 5 pp 575ndash5851996
[5] P Yang Z Zhang H Zhou et al ldquoClinical patterns andcharacteristics of uveitis in a tertiary center for uveitis in ChinardquoCurrent Eye Research vol 30 no 11 pp 943ndash948 2005
[6] J T Gran and J F Skomsvoll ldquoThe outcome of ankylosingspondylitis a study of 100 patientsrdquo British Journal of Rheuma-tology vol 36 no 7 pp 766ndash771 1997
[7] L P Robertson andM J Davis ldquoA longitudinal study of diseaseactivity and functional status in a hospital cohort of patientswith ankylosing spondylitisrdquo Rheumatology vol 43 no 12 pp1565ndash1568 2004
[8] J H Chang P J McCluskey and D Wakefield ldquoAcute anterioruveitis and HLA-B27rdquo Survey of Ophthalmology vol 50 no 4pp 364ndash388 2005
[9] M Huhtinen and A Karma ldquoHLA-B27 typing in the categori-sation of uveitis in a HLA-B27 rich populationrdquo British Journalof Ophthalmology vol 84 no 4 pp 413ndash416 2000
[10] S Torres S Borges and A Artiles ldquoHLA-B27 and clinicalfeatures of acute anterior uveitis in Cubardquo Ocular Immunologyand Inflammation vol 21 no 2 pp 119ndash123 2013
[11] R Saari R Lahti K M Saari et al ldquoFrequency of rheumaticdiseases in patients with acute anterior uveitisrdquo ScandinavianJournal of Rheumatology vol 11 no 2 pp 121ndash123 1982
[12] A B Beckingsale J Davies J M Gibson and A RalphRosenthal ldquoAcute anterior uveitis ankylosing spondylitis backpain and HLA-B27rdquo British Journal of Ophthalmology vol 68no 10 pp 741ndash745 1984
[13] G P Thomas and M A Brown ldquoGenetics and genomics ofankylosing spondylitisrdquo Immunological Reviews vol 233 no 1pp 162ndash180 2010
[14] P C Robinson and M A Brown ldquoGenetics of ankylosingspondylitisrdquoMolecular Immunology vol 57 no 1 pp 2ndash11 2014
[15] C Lopez-Larrea K Sujirachato N K Mehra et al ldquoHLA-B27 subtypes in Asian patients with ankylosing spondylitisEvidence for new associationsrdquo Tissue Antigens vol 45 no 3pp 169ndash176 1995
[16] P C Robinson T A M Claushuis A Cortes et al ldquoGeneticdissection of acute anterior uveitis reveals similarities and dif-ferences in associations observed with ankylosing spondylitisrdquoArthritis and Rheumatology vol 67 no 1 pp 140ndash151 2015
[17] H Li S Hou H Yu et al ldquoAssociation of genetic variationsin TNFSF15 with acute anterior uveitis in Chinese HanrdquoInvestigative Ophthalmology and Visual Science vol 56 no 8pp 4605ndash4610 2015
[18] Q Xiang L Chen J Fang et al ldquoTNF receptor-associated factor5 gene confers genetic predisposition to acute anterior uveitisand pediatric uveitisrdquo Arthritis Research and Therapy vol 15no 5 article R113 2013
[19] H Yu Y Liu L Zhang et al ldquoFoxO1 gene confers genetic predis-position to acute anterior uveitis with ankylosing spondylitisrdquoInvestigative Ophthalmology amp Visual Science vol 55 no 12 pp7970ndash7974 2014
[20] T M Martin L Bye N Modi et al ldquoGenotype analysis ofpolymorphisms in autoimmune susceptibility genes CTLA-4and PTPN22 in an Acute anterior uveitis cohortrdquo MolecularVision vol 15 pp 208ndash212 2009
[21] Q Zhang J Qi S Hou et al ldquoA functional variant of PTPN22confers risk for Vogt-Koyanagi-Harada syndrome but not forankylosing spondylitisrdquo PLoS ONE vol 9 no 5 article e969432014
[22] S-S Zhao J-WHu JWang X-J Lou and L-L Zhou ldquoInversecorrelation between CD4+CD25 highCD127lowminus regulatory T-cells and serum immunoglobulin A in patients with new-onset ankylosing spondylitisrdquo Journal of International MedicalResearch vol 39 no 5 pp 1968ndash1974 2011
[23] W Zou Z Wu X Xiang S Sun and J Zhang ldquoThe expressionand significance of T helper cell subsets and regulatory Tcells CD4+CD25+ in peripheral blood of patients with humanleukocyte antigen B27-positive acute anterior uveitisrdquo GraefersquosArchive for Clinical and Experimental Ophthalmology vol 252no 4 pp 665ndash672 2014
BioMed Research International 9
[24] C Wang Q Liao Y Hu and D Zhong ldquoT lymphocyte subsetimbalances in patients contribute to ankylosing spondylitisrdquoExperimental and Therapeutic Medicine vol 9 no 1 pp 250ndash256 2015
[25] A Tedgui and Z Mallat ldquoCytokines in atherosclerosispathogenic and regulatory pathwaysrdquoPhysiological Reviews vol86 no 2 pp 515ndash581 2006
[26] P J Murray ldquoThe JAK-STAT signaling pathway Input andoutput integrationrdquo Journal of Immunology vol 178 no 5 pp2623ndash2629 2007
[27] A N Mathur H-C Chang D G Zisoulis et al ldquoStat3 andStat4 direct development of IL-17-secreting Th cellsrdquo Journal ofImmunology vol 178 no 8 pp 4901ndash4907 2007
[28] G K Hansson ldquoRegulation of immune mechanisms inatherosclerosisrdquo Annals of the New York Academy of Sciencesvol 947 pp 157ndash165 2001
[29] T Shea-Donohue A Fasano A Smith and A Zhao ldquoEntericpathogens and gut function role of cytokines and STATsrdquo GutMicrobes vol 1 no 5 pp 316ndash324 2010
[30] T Taniguchi K Ogasawara A Takaoka and N Tanaka ldquoIRFfamily of transcription factors as regulators of host defenserdquoAnnual Review of Immunology vol 19 pp 623ndash655 2001
[31] M Lohoff D Ferrick H-W Mittrucker et al ldquoInterferonregulatory factor-1 is required for a T helper 1 immune responsein vivordquo Immunity vol 6 no 6 pp 681ndash689 1997
[32] S Taki T Sato K Ogasawara et al ldquoMultistage regulation ofTh1-type immune responses by the transcription factor IRF-1rdquoImmunity vol 6 no 6 pp 673ndash679 1997
[33] C Bogdan ldquoNitric oxide and the immune responserdquo NatureImmunology vol 2 no 10 pp 907ndash916 2001
[34] S Brahmachari and K Pahan ldquoMyelin basic protein primingreduces the expression of Foxp3 in T cells via nitric oxiderdquoTheJournal of Immunology vol 184 no 4 pp 1799ndash1809 2010
[35] S-W Lee H Choi S-Y Eun S Fukuyama and M CroftldquoNitric oxide modulates TGF-120573-directive signals to suppressFoxp3 + regulatory T cell differentiation and potentiate Th1developmentrdquo Journal of Immunology vol 186 no 12 pp 6972ndash6980 2011
[36] D A Jabs R B Nussenblatt J T Rosenbaum and Standard-ization of Uveitis Nomenclature Working G ldquoStandardizationof uveitis nomenclature for reporting clinical data Resultsof the First International Workshoprdquo American Journal ofOphthalmology vol 140 pp 509ndash516 2005
[37] S Van Der Linden H A Valkenburg and A Cats ldquoEvaluationof diagnostic criteria for ankylosing spondylitis A proposal formodification of the New York criteriardquo Arthritis and Rheuma-tism vol 27 no 4 pp 361ndash368 1984
[38] D Ellinghaus E Ellinghaus R P Nair et al ldquoCombined anal-ysis of genome-wide association studies for Crohn disease andpsoriasis identifies seven shared susceptibility locirdquo AmericanJournal of Human Genetics vol 90 no 4 pp 636ndash647 2012
[39] G Fortunato G Calcagno V Bresciamorra et al ldquoMultiplesclerosis and hepatitis C virus infection are associated withsingle nucleotide polymorphisms in interferon pathway genesrdquoJournal of Interferon and Cytokine Research vol 28 no 3 pp141ndash152 2008
[40] W-H Hahn J-S Suh S H Cho B-S Cho and S-D KimldquoPolymorphisms of signal transducers and activators of tran-scription 1 and 4 (STAT1 and STAT4) contribute to progressionof childhood IgA nephropathyrdquo Cytokine vol 50 no 1 pp 69ndash74 2010
[41] S Hou J Qi Q Zhang et al ldquoGenetic variants in the JAK1 geneconfer higher risk of Behcetrsquos disease with ocular involvementin Han ChineserdquoHumanGenetics vol 132 no 9 pp 1049ndash10582013
[42] K Hu S Hou F Li Q Xiang A Kijlstra and P YangldquoJAK1 but not JAK2 and STAT3 confers susceptibility toVogt-Koyanagi-Harada (VKH) syndrome in a Han Chinesepopulationrdquo Investigative Ophthalmology and Visual Sciencevol 54 no 5 pp 3360ndash3365 2013
[43] A Cortes J Hadler J P Pointon et al ldquoIdentification ofmultiple risk variants for ankylosing spondylitis through high-density genotyping of immune-related locirdquo Nature Geneticsvol 45 no 7 pp 730ndash738 2013
[44] P A Shaiq P E Stuart A Latif et al ldquoGenetic associations ofpsoriasis in a Pakistani populationrdquo British Journal of Derma-tology vol 169 no 2 pp 406ndash411 2013
[45] S-K Yang Y Jung H Kim M Hong B D Ye and KSong ldquoAssociation of FCGR2A JAK2 or HNF4A variants withulcerative colitis in Koreansrdquo Digestive and Liver Disease vol43 no 11 pp 856ndash861 2011
[46] M Garcıa-Bermudez R Lopez-Mejıas F Genre et al ldquoLackof association between JAK3 gene polymorphisms and cardio-vascular disease in Spanish patients with rheumatoid arthritisrdquoBioMed Research International vol 2015 Article ID 318364 11pages 2015
[47] R Lopez-Mejıas M Garcıa-Bermudez C Gonzalez-Juanateyet al ldquoNFKB1-94ATTG insdel polymorphism (rs28362491) isassociated with cardiovascular disease in patients with rheuma-toid arthritisrdquo Atherosclerosis vol 224 no 2 pp 426ndash429 2012
[48] H J Park J W Kim B-S Cho and J-H Chung ldquoAssociationof BH3 interacting domain death agonist (BID) gene polymor-phisms with proteinuria of immunoglobulin A nephropathyrdquoScandinavian Journal of Clinical and Laboratory Investigationvol 74 no 4 pp 329ndash335 2014
Submit your manuscripts athttpwwwhindawicom
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Anatomy Research International
PeptidesInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporation httpwwwhindawicom
International Journal of
Volume 2014
Zoology
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Molecular Biology International
GenomicsInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
BioinformaticsAdvances in
Marine BiologyJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Signal TransductionJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
BioMed Research International
Evolutionary BiologyInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Biochemistry Research International
ArchaeaHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Genetics Research International
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Advances in
Virolog y
Hindawi Publishing Corporationhttpwwwhindawicom
Nucleic AcidsJournal of
Volume 2014
Stem CellsInternational
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Enzyme Research
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
International Journal of
Microbiology
BioMed Research International 3
Table 1 PCR primer sequences and restriction enzymes
SNP Primers Restriction enzyme
rs310241 51015840 AACCACCAGCTCAACATTCCTAG 3101584051015840 CAGCCAGGTCTCCCGTAGG 31015840 BseDI
rs6718902 51015840 CGGACAAAAGCATGCACTAGA 3101584051015840 CCACCACCATTAATAGGTGACTTTA 31015840 DraI
rs2297518 51015840 TGAGCTCTTTCAGCATGAAGATC 3101584051015840 CTTCCGTGGTGGGCTGTG 31015840 TaqI
rs10758669 51015840 TGATGTAGAGACAAGGACATGCTGAGGTAC 3101584051015840 GCCAAAAGACAAAGGCAAGGGG 31015840 BanI
rs10975003 51015840 GGCCAGTCAAGAAAAAACCAGTT 3101584051015840 TTCGGAGTCTTGTCTGAGCATGT 31015840 HpaI
rs4795067 51015840 GCACTCATTCATTCATGCAAACATA 3101584051015840 GGCAGAACTTGAACCCCAGCT 31015840 NdeI
rs1547550 51015840 CTTCTCTAGGAGGCCCAGCA 3101584051015840 TGGGCACCACGATATGAGAG 31015840 BstMAI
rs2066802 51015840 ATTCCTGGAGCAGGTTCACAAG 3101584051015840 AAACATGGCCCCAAGTCACT 31015840 HindIII
method considering multiple tests Statistical significancelevel was defined as a corrected 119875 value lt 005 Statisticalanalyses were performed using SPSS version 170 (SPSS IncChicago IL USA)
3 Results
31 Clinical Features of AAU Patients The detailed clinicalfeatures and demographic characteristics of the AAUpatientsare presented in Table 2 All 929 AAU patients include 569(612) males and 360 (388) females The 714 controlsubjects consisted of 428 (599) males and 286 (401)females The average age was 398 plusmn 123 in AAU patientsand 395 plusmn 108 in the controls respectively There were nosignificant differences in age and gender between the casesand controls In addition 546 AAU patients (689) wereHLA-B27-positive whereas 246 AAU patients (311) wereHLA-B27-negative
32 The Genotype and Allele Frequency Distribution of theTested SNPs in AAU Eleven SNPs of the JAK-STAT signalingpathway genes (JAK1 JAK2 STAT1 IRF1 and NOS2) weresuccessfully genotyped There were no significant deviationsof HWE in either the cases or controls We did not observesignificant differences in the genotype and allele distribu-tions of any of the SNPs between the AAU patients andcontrol subjects after Bonferroni correction (see Supplemen-tary Table S1 in Supplementary Material available onlineat httpdxdoiorg10115520165896906) Further stratifiedanalyses of gender AS andHLA-B27 status showed a bound-ary significant association of two SNPs (rs10975003 andrs10758669) of JAK2 with AAU In female AS-positive AAUpatients there was a decreased frequency of the TT genotypeof rs10975003 compared to the female controls (OR = 055119875 = 159 times 10minus3 119875Bonferroni = 0052 Table 3) whereas nosignificant differences were observed in the genotype andallele frequencies of the other ten SNPs (119875Bonferroni gt 005Table 3) Similarly there were no significant differences in
Table 2 Clinical characteristics of the investigated subjects
Clinical features Total PercentageAAU patients 929 100Mean age plusmn SD (years) 398 plusmn 123AAU with AS 443 477AAU without AS 486 523AAU male 569 612AAU female 360 388AAU with AS (male) 326 (443 tested) 736AAU with AS (female) 117 (443 tested) 264AAU without AS (male) 243 (486 tested) 50AAU without AS (female) 243 (486 tested) 50HLA-B27+ AAU 546 (792 tested) 689HLA-B27+ AAU AS+ 348 (428 tested) 813HLA-B27+ AAU ASminus 198 (364 tested) 544Control 714 100Mean age plusmn SD (years) 395 plusmn 108Male 428 599Female 286 401
the genotype and allele frequencies of the SNPs between themale AAU patients and male controls (119875Bonferroni gt 005Supplementary Table S2)
In addition an increased frequency of theAC genotype inrs10758669 was observed in HLA-B27-positive AAU patientscompared to the healthy controls (OR = 144119875 = 162times10minus3119875Bonferroni = 0053 Table 4) whereas there were no significantdifferences in the genotype and allele frequencies of the otherten SNPs between the HLA-B27-positive AAU patients andthe control subjects (119875Bonferroni gt 005 Table 4)
However an increased frequency of the rs10758669ACgenotype was observed in HLA-B27-positive AS-positiveAAU patients compared to healthy controls (OR = 149 119875 =256times10minus3119875Bonferroni = 0084 Table 5) whereas no significant
4 BioMed Research InternationalTa
ble3Alleleandgeno
type
frequ
encies
infemaleA
AUpatie
ntsa
ndfemalec
ontro
ls
Gene
SNP
Alleleandgeno
type
AAU+AS+
(female)
AAU+ASminus
(female)
Con
trols(fe
male)119875(A
S+)
Pc(A
S+)
OR(95
CI)119875(A
Sminus)
Pc(A
Sminus)
OR(95
CI)
JAK1
rs310241
C80
(333
)138(296
)
156(287
)019
NS
124(090ndash
172)
074
NS
105(080ndash
137)
CC12
(100
)14
(60)
17(62)
019
NS
167(077ndash361)
091
NS
096
(046ndash
200)
CT56
(467
)110(472
)
122(449
)074
NS
108(070ndash
166)
060
NS
110(077ndash15
6)TT
52(433
)109(468
)133(489
)031
NS
080
(052
ndash123)
064
NS
092
(065ndash13
0)
rs2780815
G159(787
)40
0(870
)
495(853
)003
NS
064
(042ndash096)
045
NS
115(080ndash
163)
GG
63(624
)175(761
)212(731
)004
NS
061
(038ndash
099)
044
NS
117(079ndash
175)
GT
33(326
)50
(217
)
71(245
)011
NS
150(091ndash245)
046
NS
086
(057ndash12
9)TT
5(50)
5(22)
7(24)
020
NS
211(065ndash680)
086
NS
090
(028ndash287)
JAK2
rs10758669
A170(739
)328(675
)
399(723
)064
NS
109(077ndash15
4)010
NS
080
(061ndash10
4)AA
59(513
)
104(428
)144(522
)088
NS
097
(063ndash14
9)003
NS
069
(049ndash
100)
AC52
(452
)120(494
)
111(40
2)
036
NS
123(079ndash
190)
004
NS
145(102ndash205)
CC4(35)
19(78
)21
(76
)013
NS
044
(015
ndash130)
093
NS
103(054ndash
197)
rs10975003
C44
(206
)117
(258
)93
(179
)
040
NS
119(080ndash
177)
283times10minus3
009
159(117ndash217
)CC
2(19)
11(48)
8(31
)052
NS
060
(013
ndash287)
032
NS
160(063ndash406)
CT40
(374
)
95(419
)
77(296
)
015
NS
142(088ndash228)
483times10minus3
NS
171(118ndash249)
TT65
(607
)121(533)
175(673
)
023
NS
075
(047ndash12
0)15
9times10minus3
0052
055
(038ndash
080)
STAT
1
rs1547550
C205(869
)411(886)
423(870
)
095
NS
099
(062ndash15
6)047
NS
116(078ndash17
1)CC
88(746
)182(784
)185(761
)075
NS
092
(055ndash15
3)055
NS
114(074
ndash175)
CG29
(246
)47
(203
)53
(218
)
056
NS
117(070ndash
196)
068
NS
091
(059ndash
142)
GG
1(08
)3(13)
5(21
)040
NS
041
(005ndash352)
056
NS
065
(015
ndash276
)
rs2066
802
C46
(204
)81
(174
)
113(212
)
080
NS
095
(065ndash14
0)013
NS
078
(057ndash10
8)CC
1(09
)6(26)
8(30)
022
NS
029
(004ndash
234)
078
NS
086
(029ndash
250)
CT44
(389
)69
(296
)
97(363
)063
NS
112(071ndash17
6)011
NS
074
(051ndash10
7)TT
68(602
)158(678
)
162(607
)093
NS
098
(063ndash15
4)010
NS
137(095ndash19
7)
rs6718902
C132(600
)259(542
)308(538
)012
NS
129(094ndash
176)
091
NS
101(079ndash129)
CC36
(327
)72
(301
)82
(287
)043
NS
121(075ndash194)
072
NS
107(074
ndash156)
CT60
(545
)115(481
)144(503
)045
NS
118(076
ndash184)
061
NS
092
(065ndash12
9)TT
14(127
)52
(218
)
60(210
)
006
NS
055
(029ndash
103)
083
NS
105(069ndash
159)
rs10199181
A51
(248
)117
(261
)148(252
)091
NS
098
(068ndash14
1)073
NS
105(079ndash
139)
AA
6(58)
20(89)
19(65)
082
NS
090
(035ndash231)
029
NS
142(074
ndash273)
AT39
(379
)
77(344
)110(374
)
094
NS
102(064ndash
162)
048
NS
088
(061ndash12
6)TT
58(563
)127(567
)165(561
)097
NS
101(064
ndash158)
090
NS
102(072ndash14
5)
IRF1
rs2070721
A86
(406
)154(339
)216(343
)010
NS
131(095ndash180)
090
NS
098
(076
ndash127)
AA
15(14
2)
26(115
)
29(92)
015
NS
163(084ndash
317)
039
NS
128(073ndash223)
AC56
(528
)102(449
)158(502
)063
NS
111(072ndash173)
023
NS
081
(058ndash114)
CC35
(330
)99
(436
)128(406
)016
NS
072
(045ndash114)
049
NS
113(080ndash
160)
NOS2
rs2297518
A52
(205
)69
(152
)74
(157
)010
NS
139(094ndash
205)
084
NS
096
(068ndash13
8)AA
3(24)
6(26)
2(08)
024
NS
283
(047ndash1717)
014
NS
318
(063ndash1590)
AG46
(362
)57
(252
)70
(297
)
020
NS
135(085ndash213)
027
NS
080
(053
ndash120)
GG
78(614
)
164(722
)164(695
)
012
NS
070
(045ndash110)
051
NS
114(077ndash17
1)
rs4795067
A173(739
)369(759
)439(778
)
024
NS
081
(057ndash115)
046
NS
090
(067ndash12
0)AA
62(530
)145(597
)
169(599
)
020
NS
075
(049ndash
116)
095
NS
099
(070ndash
140)
AG49
(419
)
79(325
)101(358)
026
NS
129(083ndash201)
043
NS
086
(060ndash
124)
GG
6(51
)19
(78
)12
(43)
070
NS
122(045ndash332)
008
NS
191(091ndash4
02)
OR=od
dsratio
95
CI=95
confi
denceinterval
Pc=119875valuea
djustedby
Bonferroni
correctio
n
BioMed Research International 5
Table 4 Allele and genotype frequencies of SNPs in patients with AAU versus control subjects stratified by HLA-B27 status
Gene SNP Allele and genotype AAUHLA-B27 Control 119875 Pc OR (95 CI)
JAK1
rs310241
C 324 (310) 392 (275) 006 NS 118 (100ndash141)CC 41 (79) 45 (63) 029 NS 127 (082ndash196)CT 242 (464) 302 (424) 016 NS 118 (094ndash148)TT 239 (457) 366 (513) 005 NS 080 (064ndash100)
rs2780815
G 878 (862) 1250 (875) 035 NS 089 (070ndash113)GG 381 (748) 548 (767) 044 NS 090 (069ndash118)GT 116 (228) 154 (216) 061 NS 107 (082ndash141)TT 12 (24) 12 (17) 040 NS 141 (063ndash317)
JAK2
rs10758669
A 705 (659) 980 (690) 010 NS 087 (073ndash103)AA 220 (411) 346 (487) 008 NS 074 (059ndash092)AC 265 (495) 288 (406) 162 times 10minus3 0053 144 (115ndash180)CC 50 (94) 76 (107) 043 NS 086 (059ndash125)
rs10975003
C 251 (245) 272 (212) 005 NS 121 (100ndash147)CC 26 (51) 28 (44) 056 NS 118 (068ndash203)CT 199 (388) 216 (335) 006 NS 126 (099ndash160)TT 287 (561) 399 (621) 003 NS 077 (061ndash098)
STAT1
rs1547550
C 937 (872) 1254 (878) 067 NS 095 (075ndash121)CC 407 (758) 555 (777) 042 NS 090 (069ndash117)CG 123 (229) 144 (202) 024 NS 118 (090ndash154)GG 7 (13) 15 (21) 029 NS 062 (025ndash152)
rs2066802
C 190 (193) 288 (207) 039 NS 091 (074ndash112)CC 15 (30) 27 (39) 044 NS 078 (041ndash148)CT 160 (325) 234 (337) 066 NS 095 (074ndash121)TT 318 (645) 434 (624) 047 NS 109 (086ndash139)
rs6718902
C 584 (565) 778 (545) 033 NS 108 (092ndash127)CC 163 (315) 209 (293) 040 NS 111 (087ndash142)CT 258 (499) 360 (504) 086 NS 098 (078ndash123)TT 96 (186) 145 (203) 045 NS 090 (067ndash119)
rs10199181
A 270 (272) 397 (278) 075 NS 097 (081ndash116)AA 43 (87) 55 (77) 055 NS 114 (075ndash173)AT 184 (371) 287 (402) 028 NS 088 (069ndash111)TT 269 (542) 372 (521) 047 NS 109 (087ndash137)
IRF1 rs2070721
A 356 (357) 487 (341) 040 NS 108(091ndash127)AA 61 (122) 74 (104) 030 NS 121 (084ndash173)AC 234 (470) 339 (474) 087 NS 098 (078ndash123)CC 203 (408) 301 (422) 063 NS 094 (075ndash119)
NOS2
rs2297518
A 194 (181) 209 (162) 021 NS 115 (093ndash142)AA 12 (23) 12 (19) 064 NS 121 (054ndash272)AG 170 (317) 185 (286) 024 NS 116 (090ndash149)GG 354 (660) 450 (695) 020 NS 085 (067ndash109)
rs4795067
A 797 (743) 1008 (761) 035 NS 092 (076ndash110)AA 293 (546) 382 (577) 033 NS 089 (071ndash123)AG 211 (394) 244 (369) 037 NS 111 (088ndash141)GG 32 (60) 36 (54) 069 NS 110 (068ndash180)
OR = odds ratio 95 CI = 95 confidence intervalPc = 119875 value adjusted by Bonferroni correction
6 BioMed Research InternationalTa
ble5Alleleandgeno
type
frequ
encies
ofSN
Psin
patientsw
ithAAU
versus
controlsub
jectsstratified
byASandHLA
-B27
status
Gene
SNP
Alleleandgeno
type
AAU+AS+
HLA
-B27
AAU+ASminus
HLA
-B27
Con
trol
119875(A
S+)
Pc(A
S+)
OR(95
CI)119875(A
Sminus)
Pc(A
Sminus)
OR(95
CI)
JAK1
rs310241
C205(316
)
119(299
)
392(275
)
005
NS
122(100ndash
149)
032
NS
113(088ndash14
5)CC
30(93)
11(55)
45(63)
009
NS
152(094ndash
245)
070
NS
087
(044ndash
172)
CT145(447
)97
(487
)302(424
)047
NS
110(085ndash14
4)010
NS
131(095ndash179)
TT149(460
)90
(457
)366(513
)
011
NS
081
(062ndash10
5)014
NS
079
(058ndash10
8)
rs2780815
G543(843
)335(896
)
1250
(875
)
005
NS
077
(059ndash
100)
028
NS
122(085ndash17
7)GG
230(714
)
151(807)
548(767
)007
NS
076
(056ndash
102)
024
NS
127(085ndash19
0)GT
83(258
)33
(176
)
154(216
)
014
NS
126(093ndash17
2)023
NS
078
(051ndash118)
TT9(28)
3(17)
12(17)
024
NS
168(070ndash
403)
094
NS
095
(027ndash342)
JAK2
rs10758669
A44
9(662
)256(653
)980(690
)
020
NS
088
(072ndash17
0)016
NS
085
(067ndash10
7)AA
139(410
)
81(413
)
346(487
)002
NS
073
(056ndash
095)
007
NS
074
(054ndash
102)
AC171(504)
94(480
)288(406
)256times10minus3
0084
149(115
ndash194)
006
NS
135(098ndash18
6)CC
29(86)
21(107
)76
(107
)028
NS
078
(050ndash
122)
100
NS
100(060ndash
167)
rs10975003
C165(257
)86
(225
)272(212
)
003
NS
129(103ndash16
1)057
NS
108(082ndash14
3)CC
18(56)
8(42)
28(44)
039
NS
131(071ndash240)
092
NS
096
(043ndash214)
CT129(402
)70
(366
)216(335
)004
NS
133(101ndash17
5)044
NS
114(082ndash16
0)TT
174(542
)113(592
)
399(621
)001
NS
072
(055ndash095)
047
NS
089
(064ndash
123)
STAT
1
rs1547550
C608(881
)329(857
)1254
(878
)
084
NS
103(078ndash13
6)026
NS
083
(060ndash
115)
CC264(765
)143(745
)555(777
)
066
NS
093
(069ndash
127)
034
NS
084
(058ndash12
1)CG
80(232
)43
(224
)144(202
)026
NS
120(088ndash16
3)050
NS
114(078ndash16
8)GG
1(03
)6(31
)15
(21
)002
NS
014
(002ndash103)
040
NS
150(058ndash393)
rs2066
802
C130(207
)60
(168
)288(207
)099
NS
100(079ndash
126)
009
NS
077
(057ndash10
5)CC
11(35)
4(22)
27(39)
077
NS
090
(044ndash
183)
029
NS
057
(020ndash
164)
CT108(344
)52
(291
)
234(337
)082
NS
103(078ndash13
7)024
NS
081
(056ndash
116)
TT195(621
)123(687
)434(624
)092
NS
099
(075ndash13
0)012
NS
132(093ndash18
8)
rs6718902
C368(577
)
216(545
)778(545
)018
NS
114(094ndash
138)
098
NS
100(080ndash
125)
CC103(323
)60
(303
)209(293
)
033
NS
115(087ndash15
3)078
NS
105(075ndash14
8)CT
162(508
)96
(485
)360(504
)091
NS
102(078ndash13
2)063
NS
093
(068ndash12
7)TT
54(169
)42
(212
)
145(203
)020
NS
080
(057ndash113)
078
NS
106(072ndash15
6)
rs10199181
A170(269
)100(278
)
397(278
)
067
NS
096
(077ndash118)
099
NS
100(077ndash12
9)AA
25(79
)18
(100
)55
(77
)091
NS
103(063ndash16
8)032
NS
133(076
ndash233
)AT
120(380
)64
(356
)287(402
)050
NS
091
(069ndash
120)
026
NS
082
(058ndash115)
TT171(541
)98
(544
)372(521
)055
NS
108(083ndash14
1)057
NS
110(079ndash
153)
IRF1
rs2070721
A221(349)
135(373
)
487(341
)073
NS
103(085ndash12
6)025
NS
115(090-14
6)AA
37(117
)
24(133
)74
(104
)053
NS
114(075ndash174)
026
NS
132(081ndash216)
AC147(464
)87
(481
)339(474
)
074
NS
096
(073ndash12
5)089
NS
102(074
ndash142)
CC133(420
)70
(386
)301(422)
095
NS
099
(076
ndash130)
040
NS
097
(062ndash12
1)
NOS2
rs2297518
A130(19
0)
64(165
)209(162
)011
NS
122(096ndash
155)
087
NS
103(076
ndash139
)AA
7(20)
5(26)
12(19)
083
NS
111(043ndash284)
053
NS
140(049ndash
402)
AG116(339
)54
(278
)
185(286
)008
NS
128(097ndash17
0)084
NS
096
(067ndash13
8)GG
219(641
)135(696
)
450(695
)
007
NS
078
(059ndash
103)
099
NS
100(071ndash14
2)
rs4795067
A512(736
)295(745
)1008
(761
)020
NS
087
(071ndash10
8)061
NS
092
(071ndash119)
AA
184(529
)109(551
)382(577
)
014
NS
082
(063ndash10
7)051
NS
090
(065ndash12
4)AG
144(414
)
77(388
)244(369
)016
NS
121(093ndash158)
060
NS
109(079ndash
151)
GG
20(57)
12(61
)36
(54)
084
NS
106(060ndash
186)
074
NS
112(057ndash220)
OR=od
dsratio
95
CI=95
confi
denceinterval
Pc=119875valuea
djustedby
Bonferroni
correctio
n
BioMed Research International 7
differences in the genotype and allele frequencies of the other10 SNPs were observed between the HLA-B27-positive AS-positiveAAUpatients and healthy controls (119875Bonferroni gt 005Table 5) In addition there were no significant differencesin the genotype and allele frequencies of the tested SNPsbetween the AS-positive AAU patients and control subjects(119875Bonferroni gt 005 Supplementary Table S3)
33 Logistic Regression Analysis of SNPs in AAU We furtherinvestigated the SNPs rs10758669 and rs10975003 using addi-tive codominant dominant and recessive genetic modelsusing a multivariate logistic regression model adjusted forage and gender We observed that the frequency of the CAgenotype of rs10758669 was significantly higher in AAUpatients which suggests that patients with the rs10758669CA genotype have increased susceptibility to AAU (468versus 406 OR = 128 119875 = 002 SupplementaryTable S4) An increased frequency of the AC genotype ofrs10758669 was also observed in HLA-B27-positive AAUpatients and AS-positive AAU patients compared to thehealthy controls (495 versus 406 OR = 143 119875 =350 times 10minus3 and 492 versus 406 OR = 136 119875 = 002Supplementary Table S4) For SNP rs10975003 the frequencyof the heterozygous CT genotype was significantly higherin female AAU patients compared to the healthy controls(404 versus 336 OR = 157 119875 = 001 SupplementaryTable S5) A similar result was observed when we combinedCT and CC to construct a dominant model (443 versus380 OR = 157 119875 = 910times 10minus3 Supplementary Table S5)However none of the observed associations for the two SNPsretained statistical significance after Bonferroni correction(119875Bonferroni gt 005) Furthermore no significance associationswere found between the other SNPs and AAU even afterstratification by gender AS and HLA-B27 status (data notshown)
4 Discussion
In this study we first investigated whether genetic poly-morphisms of JAK-STAT signaling pathway genes includingJAK1 JAK2 STAT1 IRF1 and NOS2 confer susceptibilityto AAU with or without AS in a Chinese Han populationOur results suggest that none of these SNPs exhibit statisti-cally different frequencies of genotypes and alleles betweenhealthy controls and AAU patients However we observeda boundary significant association for two SNPs (rs10975003and rs10758669) of JAK2 by stratification analysis by genderand HLA-B27 status
We highlighted two issues at the time of study designto obtain unbiased association results First we followedstrict criteria for the diagnosis of AAU patients Patients withAAU were diagnosed as previously described by Jabs et al[36] and patients with AS were diagnosed with the modifiedNew York Criteria [37] In addition the AAU patients andhealthy controls were strictly matched by ethnicity and ageto avoid a possible influence of population stratificationFurthermore we only enrolled controls who had detailedhistories and physical examinations and excluded controlswith any autoimmune or immune-related diseases Finally
20 of the samples were randomly chosen and analyzed bydirect sequencing and the results of different genotypingmethods were consistent
AAU is defined as inflammation confined to the anteriorsegment of the eye that involves the iris and anterior part ofthe ciliary body The HLA-B27 is considered to be stronglyassociated with both AAU and AS [5 8 10 14] Recentgenetic studies have revealed that five candidate genes inthe JAK-STAT signaling pathway were considered geneticpredisposing factors for different autoimmune-mediated dis-eases [38ndash45] SNPs (rs10758669 and 10975003) in JAK2 areconsidered susceptibility factors for Crohnrsquos disease (CD) inthe German population and ulcerative colitis (UC) in theKorean population [38 45] One SNP (rs2070721) in IRF1 andSNPs (rs2297518 and rs4795067) in NOS2 are also associatedwith autoimmune diseases such as multiple sclerosis (MS)in Italy AS in Europe and psoriasis in Pakistan [39 43 44]Four SNPs (rs6718902 rs10199181 rs2066802 and rs1547550)in the STAT1 gene were observed to be associated with MSin Italy and IgA nephropathy (IgAN) in Korea [39 40] Inaddition an association was found between SNPs (rs2780815rs310241 rs3790532 rs310230 and rs310236) in JAK1 andBehcet disease (BD) as well as Vogt-Koyanagi-Harada (VKH)syndrome [41 42] two other common uveitis entities inChina There has been no report of associations betweenJAK-STAT signaling pathway genes and AAU and thus weperformed this case-control study to detect whether the fivecandidate genes were associated with AAU in a Chinese Hanpopulation Our results showed that there were no significantassociations between the genetic polymorphisms of the fivecandidate genes in the JAK-STAT signaling pathway andAAUThese results are not consistent with those observed forother autoimmune diseases reported in German Europeanand some other Asian populations [38 43 45] This discrep-ancy may be attributable to differences in the etiology andpathogenesis of AAU compared with BD VKH syndromeand autoimmune-mediated diseases
Consistent with our results a recent study also reportedno significant association of JAK-STAT signaling pathwaygene polymorphisms with rheumatoid arthritis stratified bythe presenceabsence of cardiovascular disease [46] Con-versely an influence of NFKB1 signaling pathway poly-morphisms on the development of cardiovascular events inpatients with rheumatoid arthritis has been observed [47]Furthermore NFKB1 signaling pathway polymorphismshave been described to play a critical role in the developmentof many autoimmune and inflammatory diseases and thusthe evaluation of the potential relationships between NFKB1polymorphisms and the development of AAU could be apromising research line for the future
There were several limitations of our studyWe had a lim-ited sample size to detect SNPs with weak effects while con-sidering multiple corrections Even for SNPs rs10758669 andrs10975003 we only had 700 power using a genetic powercalculator [48] In addition our samples were restricted to theHan Chinese population and all patients were enrolled fromthe ophthalmology department Further studies with a largersample size and other ethnic populations as well as patientsenrolled from multiple sources are warranted to confirm our
8 BioMed Research International
findings Additionally we only focused on eleven SNPs in theJAK-STAT pathway and it is possible that other unknownSNPs might be associated with AAU risk
In conclusion this study reveals that genetic polymor-phisms of JAK-STAT pathway genes including JAK1 JAK2STAT1 IRF1 andNOS2 may not be involved in susceptibilityto AAU risk in the Han Chinese population
Competing Interests
The authors declare that there is no conflict of interestsregarding the publication of this paper
Authorsrsquo Contributions
Ling Cheng and Hongsong Yu contributed equally to thiswork
Acknowledgments
The authors would like to thank Professor Peizeng Yang forhis great help in our study They are also grateful to alldonors enrolled in this study This work was supported bythe National Natural Science Foundation Project (8120067881670844) Fundamental and Advanced Research Programof Chongqing (cstc2015jcyjA10022) Science and TechnologyProject of Chongqing Municipal Education Commission(KJ1500236) Scientific Research Program of Science andTechnology Commission of Yuzhong District of Chongqing(20150102) and National Key Clinical Specialties Construc-tion Program of China
References
[1] D-K Hwang Y-J Chou C-Y Pu and P Chou ldquoEpidemi-ology of uveitis among the Chinese population in Taiwan APopulation-Based Studyrdquo Ophthalmology vol 119 no 11 pp2371ndash2376 2012
[2] K Nakao and N Ohba ldquoPrevalence of endogenous uveitis inKagoshima Prefecture Southwest JapanrdquoNipponGankaGakkaiZasshi vol 100 no 2 pp 150ndash155 1996
[3] D C Gritz and I GWong ldquoIncidence and prevalence of uveitisin Northern California the Northern California Epidemiologyof Uveitis Studyrdquo Ophthalmology vol 111 no 3 pp 491ndash5002004
[4] R B Nussenblatt and I Gery ldquoExperimental autoimmuneuveitis and its relationship to clinical ocular inflammatorydiseaserdquo Journal of Autoimmunity vol 9 no 5 pp 575ndash5851996
[5] P Yang Z Zhang H Zhou et al ldquoClinical patterns andcharacteristics of uveitis in a tertiary center for uveitis in ChinardquoCurrent Eye Research vol 30 no 11 pp 943ndash948 2005
[6] J T Gran and J F Skomsvoll ldquoThe outcome of ankylosingspondylitis a study of 100 patientsrdquo British Journal of Rheuma-tology vol 36 no 7 pp 766ndash771 1997
[7] L P Robertson andM J Davis ldquoA longitudinal study of diseaseactivity and functional status in a hospital cohort of patientswith ankylosing spondylitisrdquo Rheumatology vol 43 no 12 pp1565ndash1568 2004
[8] J H Chang P J McCluskey and D Wakefield ldquoAcute anterioruveitis and HLA-B27rdquo Survey of Ophthalmology vol 50 no 4pp 364ndash388 2005
[9] M Huhtinen and A Karma ldquoHLA-B27 typing in the categori-sation of uveitis in a HLA-B27 rich populationrdquo British Journalof Ophthalmology vol 84 no 4 pp 413ndash416 2000
[10] S Torres S Borges and A Artiles ldquoHLA-B27 and clinicalfeatures of acute anterior uveitis in Cubardquo Ocular Immunologyand Inflammation vol 21 no 2 pp 119ndash123 2013
[11] R Saari R Lahti K M Saari et al ldquoFrequency of rheumaticdiseases in patients with acute anterior uveitisrdquo ScandinavianJournal of Rheumatology vol 11 no 2 pp 121ndash123 1982
[12] A B Beckingsale J Davies J M Gibson and A RalphRosenthal ldquoAcute anterior uveitis ankylosing spondylitis backpain and HLA-B27rdquo British Journal of Ophthalmology vol 68no 10 pp 741ndash745 1984
[13] G P Thomas and M A Brown ldquoGenetics and genomics ofankylosing spondylitisrdquo Immunological Reviews vol 233 no 1pp 162ndash180 2010
[14] P C Robinson and M A Brown ldquoGenetics of ankylosingspondylitisrdquoMolecular Immunology vol 57 no 1 pp 2ndash11 2014
[15] C Lopez-Larrea K Sujirachato N K Mehra et al ldquoHLA-B27 subtypes in Asian patients with ankylosing spondylitisEvidence for new associationsrdquo Tissue Antigens vol 45 no 3pp 169ndash176 1995
[16] P C Robinson T A M Claushuis A Cortes et al ldquoGeneticdissection of acute anterior uveitis reveals similarities and dif-ferences in associations observed with ankylosing spondylitisrdquoArthritis and Rheumatology vol 67 no 1 pp 140ndash151 2015
[17] H Li S Hou H Yu et al ldquoAssociation of genetic variationsin TNFSF15 with acute anterior uveitis in Chinese HanrdquoInvestigative Ophthalmology and Visual Science vol 56 no 8pp 4605ndash4610 2015
[18] Q Xiang L Chen J Fang et al ldquoTNF receptor-associated factor5 gene confers genetic predisposition to acute anterior uveitisand pediatric uveitisrdquo Arthritis Research and Therapy vol 15no 5 article R113 2013
[19] H Yu Y Liu L Zhang et al ldquoFoxO1 gene confers genetic predis-position to acute anterior uveitis with ankylosing spondylitisrdquoInvestigative Ophthalmology amp Visual Science vol 55 no 12 pp7970ndash7974 2014
[20] T M Martin L Bye N Modi et al ldquoGenotype analysis ofpolymorphisms in autoimmune susceptibility genes CTLA-4and PTPN22 in an Acute anterior uveitis cohortrdquo MolecularVision vol 15 pp 208ndash212 2009
[21] Q Zhang J Qi S Hou et al ldquoA functional variant of PTPN22confers risk for Vogt-Koyanagi-Harada syndrome but not forankylosing spondylitisrdquo PLoS ONE vol 9 no 5 article e969432014
[22] S-S Zhao J-WHu JWang X-J Lou and L-L Zhou ldquoInversecorrelation between CD4+CD25 highCD127lowminus regulatory T-cells and serum immunoglobulin A in patients with new-onset ankylosing spondylitisrdquo Journal of International MedicalResearch vol 39 no 5 pp 1968ndash1974 2011
[23] W Zou Z Wu X Xiang S Sun and J Zhang ldquoThe expressionand significance of T helper cell subsets and regulatory Tcells CD4+CD25+ in peripheral blood of patients with humanleukocyte antigen B27-positive acute anterior uveitisrdquo GraefersquosArchive for Clinical and Experimental Ophthalmology vol 252no 4 pp 665ndash672 2014
BioMed Research International 9
[24] C Wang Q Liao Y Hu and D Zhong ldquoT lymphocyte subsetimbalances in patients contribute to ankylosing spondylitisrdquoExperimental and Therapeutic Medicine vol 9 no 1 pp 250ndash256 2015
[25] A Tedgui and Z Mallat ldquoCytokines in atherosclerosispathogenic and regulatory pathwaysrdquoPhysiological Reviews vol86 no 2 pp 515ndash581 2006
[26] P J Murray ldquoThe JAK-STAT signaling pathway Input andoutput integrationrdquo Journal of Immunology vol 178 no 5 pp2623ndash2629 2007
[27] A N Mathur H-C Chang D G Zisoulis et al ldquoStat3 andStat4 direct development of IL-17-secreting Th cellsrdquo Journal ofImmunology vol 178 no 8 pp 4901ndash4907 2007
[28] G K Hansson ldquoRegulation of immune mechanisms inatherosclerosisrdquo Annals of the New York Academy of Sciencesvol 947 pp 157ndash165 2001
[29] T Shea-Donohue A Fasano A Smith and A Zhao ldquoEntericpathogens and gut function role of cytokines and STATsrdquo GutMicrobes vol 1 no 5 pp 316ndash324 2010
[30] T Taniguchi K Ogasawara A Takaoka and N Tanaka ldquoIRFfamily of transcription factors as regulators of host defenserdquoAnnual Review of Immunology vol 19 pp 623ndash655 2001
[31] M Lohoff D Ferrick H-W Mittrucker et al ldquoInterferonregulatory factor-1 is required for a T helper 1 immune responsein vivordquo Immunity vol 6 no 6 pp 681ndash689 1997
[32] S Taki T Sato K Ogasawara et al ldquoMultistage regulation ofTh1-type immune responses by the transcription factor IRF-1rdquoImmunity vol 6 no 6 pp 673ndash679 1997
[33] C Bogdan ldquoNitric oxide and the immune responserdquo NatureImmunology vol 2 no 10 pp 907ndash916 2001
[34] S Brahmachari and K Pahan ldquoMyelin basic protein primingreduces the expression of Foxp3 in T cells via nitric oxiderdquoTheJournal of Immunology vol 184 no 4 pp 1799ndash1809 2010
[35] S-W Lee H Choi S-Y Eun S Fukuyama and M CroftldquoNitric oxide modulates TGF-120573-directive signals to suppressFoxp3 + regulatory T cell differentiation and potentiate Th1developmentrdquo Journal of Immunology vol 186 no 12 pp 6972ndash6980 2011
[36] D A Jabs R B Nussenblatt J T Rosenbaum and Standard-ization of Uveitis Nomenclature Working G ldquoStandardizationof uveitis nomenclature for reporting clinical data Resultsof the First International Workshoprdquo American Journal ofOphthalmology vol 140 pp 509ndash516 2005
[37] S Van Der Linden H A Valkenburg and A Cats ldquoEvaluationof diagnostic criteria for ankylosing spondylitis A proposal formodification of the New York criteriardquo Arthritis and Rheuma-tism vol 27 no 4 pp 361ndash368 1984
[38] D Ellinghaus E Ellinghaus R P Nair et al ldquoCombined anal-ysis of genome-wide association studies for Crohn disease andpsoriasis identifies seven shared susceptibility locirdquo AmericanJournal of Human Genetics vol 90 no 4 pp 636ndash647 2012
[39] G Fortunato G Calcagno V Bresciamorra et al ldquoMultiplesclerosis and hepatitis C virus infection are associated withsingle nucleotide polymorphisms in interferon pathway genesrdquoJournal of Interferon and Cytokine Research vol 28 no 3 pp141ndash152 2008
[40] W-H Hahn J-S Suh S H Cho B-S Cho and S-D KimldquoPolymorphisms of signal transducers and activators of tran-scription 1 and 4 (STAT1 and STAT4) contribute to progressionof childhood IgA nephropathyrdquo Cytokine vol 50 no 1 pp 69ndash74 2010
[41] S Hou J Qi Q Zhang et al ldquoGenetic variants in the JAK1 geneconfer higher risk of Behcetrsquos disease with ocular involvementin Han ChineserdquoHumanGenetics vol 132 no 9 pp 1049ndash10582013
[42] K Hu S Hou F Li Q Xiang A Kijlstra and P YangldquoJAK1 but not JAK2 and STAT3 confers susceptibility toVogt-Koyanagi-Harada (VKH) syndrome in a Han Chinesepopulationrdquo Investigative Ophthalmology and Visual Sciencevol 54 no 5 pp 3360ndash3365 2013
[43] A Cortes J Hadler J P Pointon et al ldquoIdentification ofmultiple risk variants for ankylosing spondylitis through high-density genotyping of immune-related locirdquo Nature Geneticsvol 45 no 7 pp 730ndash738 2013
[44] P A Shaiq P E Stuart A Latif et al ldquoGenetic associations ofpsoriasis in a Pakistani populationrdquo British Journal of Derma-tology vol 169 no 2 pp 406ndash411 2013
[45] S-K Yang Y Jung H Kim M Hong B D Ye and KSong ldquoAssociation of FCGR2A JAK2 or HNF4A variants withulcerative colitis in Koreansrdquo Digestive and Liver Disease vol43 no 11 pp 856ndash861 2011
[46] M Garcıa-Bermudez R Lopez-Mejıas F Genre et al ldquoLackof association between JAK3 gene polymorphisms and cardio-vascular disease in Spanish patients with rheumatoid arthritisrdquoBioMed Research International vol 2015 Article ID 318364 11pages 2015
[47] R Lopez-Mejıas M Garcıa-Bermudez C Gonzalez-Juanateyet al ldquoNFKB1-94ATTG insdel polymorphism (rs28362491) isassociated with cardiovascular disease in patients with rheuma-toid arthritisrdquo Atherosclerosis vol 224 no 2 pp 426ndash429 2012
[48] H J Park J W Kim B-S Cho and J-H Chung ldquoAssociationof BH3 interacting domain death agonist (BID) gene polymor-phisms with proteinuria of immunoglobulin A nephropathyrdquoScandinavian Journal of Clinical and Laboratory Investigationvol 74 no 4 pp 329ndash335 2014
Submit your manuscripts athttpwwwhindawicom
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
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PeptidesInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
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International Journal of
Volume 2014
Zoology
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Molecular Biology International
GenomicsInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
BioinformaticsAdvances in
Marine BiologyJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Signal TransductionJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
BioMed Research International
Evolutionary BiologyInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Biochemistry Research International
ArchaeaHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Genetics Research International
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
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Volume 2014
Stem CellsInternational
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
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Enzyme Research
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
International Journal of
Microbiology
4 BioMed Research InternationalTa
ble3Alleleandgeno
type
frequ
encies
infemaleA
AUpatie
ntsa
ndfemalec
ontro
ls
Gene
SNP
Alleleandgeno
type
AAU+AS+
(female)
AAU+ASminus
(female)
Con
trols(fe
male)119875(A
S+)
Pc(A
S+)
OR(95
CI)119875(A
Sminus)
Pc(A
Sminus)
OR(95
CI)
JAK1
rs310241
C80
(333
)138(296
)
156(287
)019
NS
124(090ndash
172)
074
NS
105(080ndash
137)
CC12
(100
)14
(60)
17(62)
019
NS
167(077ndash361)
091
NS
096
(046ndash
200)
CT56
(467
)110(472
)
122(449
)074
NS
108(070ndash
166)
060
NS
110(077ndash15
6)TT
52(433
)109(468
)133(489
)031
NS
080
(052
ndash123)
064
NS
092
(065ndash13
0)
rs2780815
G159(787
)40
0(870
)
495(853
)003
NS
064
(042ndash096)
045
NS
115(080ndash
163)
GG
63(624
)175(761
)212(731
)004
NS
061
(038ndash
099)
044
NS
117(079ndash
175)
GT
33(326
)50
(217
)
71(245
)011
NS
150(091ndash245)
046
NS
086
(057ndash12
9)TT
5(50)
5(22)
7(24)
020
NS
211(065ndash680)
086
NS
090
(028ndash287)
JAK2
rs10758669
A170(739
)328(675
)
399(723
)064
NS
109(077ndash15
4)010
NS
080
(061ndash10
4)AA
59(513
)
104(428
)144(522
)088
NS
097
(063ndash14
9)003
NS
069
(049ndash
100)
AC52
(452
)120(494
)
111(40
2)
036
NS
123(079ndash
190)
004
NS
145(102ndash205)
CC4(35)
19(78
)21
(76
)013
NS
044
(015
ndash130)
093
NS
103(054ndash
197)
rs10975003
C44
(206
)117
(258
)93
(179
)
040
NS
119(080ndash
177)
283times10minus3
009
159(117ndash217
)CC
2(19)
11(48)
8(31
)052
NS
060
(013
ndash287)
032
NS
160(063ndash406)
CT40
(374
)
95(419
)
77(296
)
015
NS
142(088ndash228)
483times10minus3
NS
171(118ndash249)
TT65
(607
)121(533)
175(673
)
023
NS
075
(047ndash12
0)15
9times10minus3
0052
055
(038ndash
080)
STAT
1
rs1547550
C205(869
)411(886)
423(870
)
095
NS
099
(062ndash15
6)047
NS
116(078ndash17
1)CC
88(746
)182(784
)185(761
)075
NS
092
(055ndash15
3)055
NS
114(074
ndash175)
CG29
(246
)47
(203
)53
(218
)
056
NS
117(070ndash
196)
068
NS
091
(059ndash
142)
GG
1(08
)3(13)
5(21
)040
NS
041
(005ndash352)
056
NS
065
(015
ndash276
)
rs2066
802
C46
(204
)81
(174
)
113(212
)
080
NS
095
(065ndash14
0)013
NS
078
(057ndash10
8)CC
1(09
)6(26)
8(30)
022
NS
029
(004ndash
234)
078
NS
086
(029ndash
250)
CT44
(389
)69
(296
)
97(363
)063
NS
112(071ndash17
6)011
NS
074
(051ndash10
7)TT
68(602
)158(678
)
162(607
)093
NS
098
(063ndash15
4)010
NS
137(095ndash19
7)
rs6718902
C132(600
)259(542
)308(538
)012
NS
129(094ndash
176)
091
NS
101(079ndash129)
CC36
(327
)72
(301
)82
(287
)043
NS
121(075ndash194)
072
NS
107(074
ndash156)
CT60
(545
)115(481
)144(503
)045
NS
118(076
ndash184)
061
NS
092
(065ndash12
9)TT
14(127
)52
(218
)
60(210
)
006
NS
055
(029ndash
103)
083
NS
105(069ndash
159)
rs10199181
A51
(248
)117
(261
)148(252
)091
NS
098
(068ndash14
1)073
NS
105(079ndash
139)
AA
6(58)
20(89)
19(65)
082
NS
090
(035ndash231)
029
NS
142(074
ndash273)
AT39
(379
)
77(344
)110(374
)
094
NS
102(064ndash
162)
048
NS
088
(061ndash12
6)TT
58(563
)127(567
)165(561
)097
NS
101(064
ndash158)
090
NS
102(072ndash14
5)
IRF1
rs2070721
A86
(406
)154(339
)216(343
)010
NS
131(095ndash180)
090
NS
098
(076
ndash127)
AA
15(14
2)
26(115
)
29(92)
015
NS
163(084ndash
317)
039
NS
128(073ndash223)
AC56
(528
)102(449
)158(502
)063
NS
111(072ndash173)
023
NS
081
(058ndash114)
CC35
(330
)99
(436
)128(406
)016
NS
072
(045ndash114)
049
NS
113(080ndash
160)
NOS2
rs2297518
A52
(205
)69
(152
)74
(157
)010
NS
139(094ndash
205)
084
NS
096
(068ndash13
8)AA
3(24)
6(26)
2(08)
024
NS
283
(047ndash1717)
014
NS
318
(063ndash1590)
AG46
(362
)57
(252
)70
(297
)
020
NS
135(085ndash213)
027
NS
080
(053
ndash120)
GG
78(614
)
164(722
)164(695
)
012
NS
070
(045ndash110)
051
NS
114(077ndash17
1)
rs4795067
A173(739
)369(759
)439(778
)
024
NS
081
(057ndash115)
046
NS
090
(067ndash12
0)AA
62(530
)145(597
)
169(599
)
020
NS
075
(049ndash
116)
095
NS
099
(070ndash
140)
AG49
(419
)
79(325
)101(358)
026
NS
129(083ndash201)
043
NS
086
(060ndash
124)
GG
6(51
)19
(78
)12
(43)
070
NS
122(045ndash332)
008
NS
191(091ndash4
02)
OR=od
dsratio
95
CI=95
confi
denceinterval
Pc=119875valuea
djustedby
Bonferroni
correctio
n
BioMed Research International 5
Table 4 Allele and genotype frequencies of SNPs in patients with AAU versus control subjects stratified by HLA-B27 status
Gene SNP Allele and genotype AAUHLA-B27 Control 119875 Pc OR (95 CI)
JAK1
rs310241
C 324 (310) 392 (275) 006 NS 118 (100ndash141)CC 41 (79) 45 (63) 029 NS 127 (082ndash196)CT 242 (464) 302 (424) 016 NS 118 (094ndash148)TT 239 (457) 366 (513) 005 NS 080 (064ndash100)
rs2780815
G 878 (862) 1250 (875) 035 NS 089 (070ndash113)GG 381 (748) 548 (767) 044 NS 090 (069ndash118)GT 116 (228) 154 (216) 061 NS 107 (082ndash141)TT 12 (24) 12 (17) 040 NS 141 (063ndash317)
JAK2
rs10758669
A 705 (659) 980 (690) 010 NS 087 (073ndash103)AA 220 (411) 346 (487) 008 NS 074 (059ndash092)AC 265 (495) 288 (406) 162 times 10minus3 0053 144 (115ndash180)CC 50 (94) 76 (107) 043 NS 086 (059ndash125)
rs10975003
C 251 (245) 272 (212) 005 NS 121 (100ndash147)CC 26 (51) 28 (44) 056 NS 118 (068ndash203)CT 199 (388) 216 (335) 006 NS 126 (099ndash160)TT 287 (561) 399 (621) 003 NS 077 (061ndash098)
STAT1
rs1547550
C 937 (872) 1254 (878) 067 NS 095 (075ndash121)CC 407 (758) 555 (777) 042 NS 090 (069ndash117)CG 123 (229) 144 (202) 024 NS 118 (090ndash154)GG 7 (13) 15 (21) 029 NS 062 (025ndash152)
rs2066802
C 190 (193) 288 (207) 039 NS 091 (074ndash112)CC 15 (30) 27 (39) 044 NS 078 (041ndash148)CT 160 (325) 234 (337) 066 NS 095 (074ndash121)TT 318 (645) 434 (624) 047 NS 109 (086ndash139)
rs6718902
C 584 (565) 778 (545) 033 NS 108 (092ndash127)CC 163 (315) 209 (293) 040 NS 111 (087ndash142)CT 258 (499) 360 (504) 086 NS 098 (078ndash123)TT 96 (186) 145 (203) 045 NS 090 (067ndash119)
rs10199181
A 270 (272) 397 (278) 075 NS 097 (081ndash116)AA 43 (87) 55 (77) 055 NS 114 (075ndash173)AT 184 (371) 287 (402) 028 NS 088 (069ndash111)TT 269 (542) 372 (521) 047 NS 109 (087ndash137)
IRF1 rs2070721
A 356 (357) 487 (341) 040 NS 108(091ndash127)AA 61 (122) 74 (104) 030 NS 121 (084ndash173)AC 234 (470) 339 (474) 087 NS 098 (078ndash123)CC 203 (408) 301 (422) 063 NS 094 (075ndash119)
NOS2
rs2297518
A 194 (181) 209 (162) 021 NS 115 (093ndash142)AA 12 (23) 12 (19) 064 NS 121 (054ndash272)AG 170 (317) 185 (286) 024 NS 116 (090ndash149)GG 354 (660) 450 (695) 020 NS 085 (067ndash109)
rs4795067
A 797 (743) 1008 (761) 035 NS 092 (076ndash110)AA 293 (546) 382 (577) 033 NS 089 (071ndash123)AG 211 (394) 244 (369) 037 NS 111 (088ndash141)GG 32 (60) 36 (54) 069 NS 110 (068ndash180)
OR = odds ratio 95 CI = 95 confidence intervalPc = 119875 value adjusted by Bonferroni correction
6 BioMed Research InternationalTa
ble5Alleleandgeno
type
frequ
encies
ofSN
Psin
patientsw
ithAAU
versus
controlsub
jectsstratified
byASandHLA
-B27
status
Gene
SNP
Alleleandgeno
type
AAU+AS+
HLA
-B27
AAU+ASminus
HLA
-B27
Con
trol
119875(A
S+)
Pc(A
S+)
OR(95
CI)119875(A
Sminus)
Pc(A
Sminus)
OR(95
CI)
JAK1
rs310241
C205(316
)
119(299
)
392(275
)
005
NS
122(100ndash
149)
032
NS
113(088ndash14
5)CC
30(93)
11(55)
45(63)
009
NS
152(094ndash
245)
070
NS
087
(044ndash
172)
CT145(447
)97
(487
)302(424
)047
NS
110(085ndash14
4)010
NS
131(095ndash179)
TT149(460
)90
(457
)366(513
)
011
NS
081
(062ndash10
5)014
NS
079
(058ndash10
8)
rs2780815
G543(843
)335(896
)
1250
(875
)
005
NS
077
(059ndash
100)
028
NS
122(085ndash17
7)GG
230(714
)
151(807)
548(767
)007
NS
076
(056ndash
102)
024
NS
127(085ndash19
0)GT
83(258
)33
(176
)
154(216
)
014
NS
126(093ndash17
2)023
NS
078
(051ndash118)
TT9(28)
3(17)
12(17)
024
NS
168(070ndash
403)
094
NS
095
(027ndash342)
JAK2
rs10758669
A44
9(662
)256(653
)980(690
)
020
NS
088
(072ndash17
0)016
NS
085
(067ndash10
7)AA
139(410
)
81(413
)
346(487
)002
NS
073
(056ndash
095)
007
NS
074
(054ndash
102)
AC171(504)
94(480
)288(406
)256times10minus3
0084
149(115
ndash194)
006
NS
135(098ndash18
6)CC
29(86)
21(107
)76
(107
)028
NS
078
(050ndash
122)
100
NS
100(060ndash
167)
rs10975003
C165(257
)86
(225
)272(212
)
003
NS
129(103ndash16
1)057
NS
108(082ndash14
3)CC
18(56)
8(42)
28(44)
039
NS
131(071ndash240)
092
NS
096
(043ndash214)
CT129(402
)70
(366
)216(335
)004
NS
133(101ndash17
5)044
NS
114(082ndash16
0)TT
174(542
)113(592
)
399(621
)001
NS
072
(055ndash095)
047
NS
089
(064ndash
123)
STAT
1
rs1547550
C608(881
)329(857
)1254
(878
)
084
NS
103(078ndash13
6)026
NS
083
(060ndash
115)
CC264(765
)143(745
)555(777
)
066
NS
093
(069ndash
127)
034
NS
084
(058ndash12
1)CG
80(232
)43
(224
)144(202
)026
NS
120(088ndash16
3)050
NS
114(078ndash16
8)GG
1(03
)6(31
)15
(21
)002
NS
014
(002ndash103)
040
NS
150(058ndash393)
rs2066
802
C130(207
)60
(168
)288(207
)099
NS
100(079ndash
126)
009
NS
077
(057ndash10
5)CC
11(35)
4(22)
27(39)
077
NS
090
(044ndash
183)
029
NS
057
(020ndash
164)
CT108(344
)52
(291
)
234(337
)082
NS
103(078ndash13
7)024
NS
081
(056ndash
116)
TT195(621
)123(687
)434(624
)092
NS
099
(075ndash13
0)012
NS
132(093ndash18
8)
rs6718902
C368(577
)
216(545
)778(545
)018
NS
114(094ndash
138)
098
NS
100(080ndash
125)
CC103(323
)60
(303
)209(293
)
033
NS
115(087ndash15
3)078
NS
105(075ndash14
8)CT
162(508
)96
(485
)360(504
)091
NS
102(078ndash13
2)063
NS
093
(068ndash12
7)TT
54(169
)42
(212
)
145(203
)020
NS
080
(057ndash113)
078
NS
106(072ndash15
6)
rs10199181
A170(269
)100(278
)
397(278
)
067
NS
096
(077ndash118)
099
NS
100(077ndash12
9)AA
25(79
)18
(100
)55
(77
)091
NS
103(063ndash16
8)032
NS
133(076
ndash233
)AT
120(380
)64
(356
)287(402
)050
NS
091
(069ndash
120)
026
NS
082
(058ndash115)
TT171(541
)98
(544
)372(521
)055
NS
108(083ndash14
1)057
NS
110(079ndash
153)
IRF1
rs2070721
A221(349)
135(373
)
487(341
)073
NS
103(085ndash12
6)025
NS
115(090-14
6)AA
37(117
)
24(133
)74
(104
)053
NS
114(075ndash174)
026
NS
132(081ndash216)
AC147(464
)87
(481
)339(474
)
074
NS
096
(073ndash12
5)089
NS
102(074
ndash142)
CC133(420
)70
(386
)301(422)
095
NS
099
(076
ndash130)
040
NS
097
(062ndash12
1)
NOS2
rs2297518
A130(19
0)
64(165
)209(162
)011
NS
122(096ndash
155)
087
NS
103(076
ndash139
)AA
7(20)
5(26)
12(19)
083
NS
111(043ndash284)
053
NS
140(049ndash
402)
AG116(339
)54
(278
)
185(286
)008
NS
128(097ndash17
0)084
NS
096
(067ndash13
8)GG
219(641
)135(696
)
450(695
)
007
NS
078
(059ndash
103)
099
NS
100(071ndash14
2)
rs4795067
A512(736
)295(745
)1008
(761
)020
NS
087
(071ndash10
8)061
NS
092
(071ndash119)
AA
184(529
)109(551
)382(577
)
014
NS
082
(063ndash10
7)051
NS
090
(065ndash12
4)AG
144(414
)
77(388
)244(369
)016
NS
121(093ndash158)
060
NS
109(079ndash
151)
GG
20(57)
12(61
)36
(54)
084
NS
106(060ndash
186)
074
NS
112(057ndash220)
OR=od
dsratio
95
CI=95
confi
denceinterval
Pc=119875valuea
djustedby
Bonferroni
correctio
n
BioMed Research International 7
differences in the genotype and allele frequencies of the other10 SNPs were observed between the HLA-B27-positive AS-positiveAAUpatients and healthy controls (119875Bonferroni gt 005Table 5) In addition there were no significant differencesin the genotype and allele frequencies of the tested SNPsbetween the AS-positive AAU patients and control subjects(119875Bonferroni gt 005 Supplementary Table S3)
33 Logistic Regression Analysis of SNPs in AAU We furtherinvestigated the SNPs rs10758669 and rs10975003 using addi-tive codominant dominant and recessive genetic modelsusing a multivariate logistic regression model adjusted forage and gender We observed that the frequency of the CAgenotype of rs10758669 was significantly higher in AAUpatients which suggests that patients with the rs10758669CA genotype have increased susceptibility to AAU (468versus 406 OR = 128 119875 = 002 SupplementaryTable S4) An increased frequency of the AC genotype ofrs10758669 was also observed in HLA-B27-positive AAUpatients and AS-positive AAU patients compared to thehealthy controls (495 versus 406 OR = 143 119875 =350 times 10minus3 and 492 versus 406 OR = 136 119875 = 002Supplementary Table S4) For SNP rs10975003 the frequencyof the heterozygous CT genotype was significantly higherin female AAU patients compared to the healthy controls(404 versus 336 OR = 157 119875 = 001 SupplementaryTable S5) A similar result was observed when we combinedCT and CC to construct a dominant model (443 versus380 OR = 157 119875 = 910times 10minus3 Supplementary Table S5)However none of the observed associations for the two SNPsretained statistical significance after Bonferroni correction(119875Bonferroni gt 005) Furthermore no significance associationswere found between the other SNPs and AAU even afterstratification by gender AS and HLA-B27 status (data notshown)
4 Discussion
In this study we first investigated whether genetic poly-morphisms of JAK-STAT signaling pathway genes includingJAK1 JAK2 STAT1 IRF1 and NOS2 confer susceptibilityto AAU with or without AS in a Chinese Han populationOur results suggest that none of these SNPs exhibit statisti-cally different frequencies of genotypes and alleles betweenhealthy controls and AAU patients However we observeda boundary significant association for two SNPs (rs10975003and rs10758669) of JAK2 by stratification analysis by genderand HLA-B27 status
We highlighted two issues at the time of study designto obtain unbiased association results First we followedstrict criteria for the diagnosis of AAU patients Patients withAAU were diagnosed as previously described by Jabs et al[36] and patients with AS were diagnosed with the modifiedNew York Criteria [37] In addition the AAU patients andhealthy controls were strictly matched by ethnicity and ageto avoid a possible influence of population stratificationFurthermore we only enrolled controls who had detailedhistories and physical examinations and excluded controlswith any autoimmune or immune-related diseases Finally
20 of the samples were randomly chosen and analyzed bydirect sequencing and the results of different genotypingmethods were consistent
AAU is defined as inflammation confined to the anteriorsegment of the eye that involves the iris and anterior part ofthe ciliary body The HLA-B27 is considered to be stronglyassociated with both AAU and AS [5 8 10 14] Recentgenetic studies have revealed that five candidate genes inthe JAK-STAT signaling pathway were considered geneticpredisposing factors for different autoimmune-mediated dis-eases [38ndash45] SNPs (rs10758669 and 10975003) in JAK2 areconsidered susceptibility factors for Crohnrsquos disease (CD) inthe German population and ulcerative colitis (UC) in theKorean population [38 45] One SNP (rs2070721) in IRF1 andSNPs (rs2297518 and rs4795067) in NOS2 are also associatedwith autoimmune diseases such as multiple sclerosis (MS)in Italy AS in Europe and psoriasis in Pakistan [39 43 44]Four SNPs (rs6718902 rs10199181 rs2066802 and rs1547550)in the STAT1 gene were observed to be associated with MSin Italy and IgA nephropathy (IgAN) in Korea [39 40] Inaddition an association was found between SNPs (rs2780815rs310241 rs3790532 rs310230 and rs310236) in JAK1 andBehcet disease (BD) as well as Vogt-Koyanagi-Harada (VKH)syndrome [41 42] two other common uveitis entities inChina There has been no report of associations betweenJAK-STAT signaling pathway genes and AAU and thus weperformed this case-control study to detect whether the fivecandidate genes were associated with AAU in a Chinese Hanpopulation Our results showed that there were no significantassociations between the genetic polymorphisms of the fivecandidate genes in the JAK-STAT signaling pathway andAAUThese results are not consistent with those observed forother autoimmune diseases reported in German Europeanand some other Asian populations [38 43 45] This discrep-ancy may be attributable to differences in the etiology andpathogenesis of AAU compared with BD VKH syndromeand autoimmune-mediated diseases
Consistent with our results a recent study also reportedno significant association of JAK-STAT signaling pathwaygene polymorphisms with rheumatoid arthritis stratified bythe presenceabsence of cardiovascular disease [46] Con-versely an influence of NFKB1 signaling pathway poly-morphisms on the development of cardiovascular events inpatients with rheumatoid arthritis has been observed [47]Furthermore NFKB1 signaling pathway polymorphismshave been described to play a critical role in the developmentof many autoimmune and inflammatory diseases and thusthe evaluation of the potential relationships between NFKB1polymorphisms and the development of AAU could be apromising research line for the future
There were several limitations of our studyWe had a lim-ited sample size to detect SNPs with weak effects while con-sidering multiple corrections Even for SNPs rs10758669 andrs10975003 we only had 700 power using a genetic powercalculator [48] In addition our samples were restricted to theHan Chinese population and all patients were enrolled fromthe ophthalmology department Further studies with a largersample size and other ethnic populations as well as patientsenrolled from multiple sources are warranted to confirm our
8 BioMed Research International
findings Additionally we only focused on eleven SNPs in theJAK-STAT pathway and it is possible that other unknownSNPs might be associated with AAU risk
In conclusion this study reveals that genetic polymor-phisms of JAK-STAT pathway genes including JAK1 JAK2STAT1 IRF1 andNOS2 may not be involved in susceptibilityto AAU risk in the Han Chinese population
Competing Interests
The authors declare that there is no conflict of interestsregarding the publication of this paper
Authorsrsquo Contributions
Ling Cheng and Hongsong Yu contributed equally to thiswork
Acknowledgments
The authors would like to thank Professor Peizeng Yang forhis great help in our study They are also grateful to alldonors enrolled in this study This work was supported bythe National Natural Science Foundation Project (8120067881670844) Fundamental and Advanced Research Programof Chongqing (cstc2015jcyjA10022) Science and TechnologyProject of Chongqing Municipal Education Commission(KJ1500236) Scientific Research Program of Science andTechnology Commission of Yuzhong District of Chongqing(20150102) and National Key Clinical Specialties Construc-tion Program of China
References
[1] D-K Hwang Y-J Chou C-Y Pu and P Chou ldquoEpidemi-ology of uveitis among the Chinese population in Taiwan APopulation-Based Studyrdquo Ophthalmology vol 119 no 11 pp2371ndash2376 2012
[2] K Nakao and N Ohba ldquoPrevalence of endogenous uveitis inKagoshima Prefecture Southwest JapanrdquoNipponGankaGakkaiZasshi vol 100 no 2 pp 150ndash155 1996
[3] D C Gritz and I GWong ldquoIncidence and prevalence of uveitisin Northern California the Northern California Epidemiologyof Uveitis Studyrdquo Ophthalmology vol 111 no 3 pp 491ndash5002004
[4] R B Nussenblatt and I Gery ldquoExperimental autoimmuneuveitis and its relationship to clinical ocular inflammatorydiseaserdquo Journal of Autoimmunity vol 9 no 5 pp 575ndash5851996
[5] P Yang Z Zhang H Zhou et al ldquoClinical patterns andcharacteristics of uveitis in a tertiary center for uveitis in ChinardquoCurrent Eye Research vol 30 no 11 pp 943ndash948 2005
[6] J T Gran and J F Skomsvoll ldquoThe outcome of ankylosingspondylitis a study of 100 patientsrdquo British Journal of Rheuma-tology vol 36 no 7 pp 766ndash771 1997
[7] L P Robertson andM J Davis ldquoA longitudinal study of diseaseactivity and functional status in a hospital cohort of patientswith ankylosing spondylitisrdquo Rheumatology vol 43 no 12 pp1565ndash1568 2004
[8] J H Chang P J McCluskey and D Wakefield ldquoAcute anterioruveitis and HLA-B27rdquo Survey of Ophthalmology vol 50 no 4pp 364ndash388 2005
[9] M Huhtinen and A Karma ldquoHLA-B27 typing in the categori-sation of uveitis in a HLA-B27 rich populationrdquo British Journalof Ophthalmology vol 84 no 4 pp 413ndash416 2000
[10] S Torres S Borges and A Artiles ldquoHLA-B27 and clinicalfeatures of acute anterior uveitis in Cubardquo Ocular Immunologyand Inflammation vol 21 no 2 pp 119ndash123 2013
[11] R Saari R Lahti K M Saari et al ldquoFrequency of rheumaticdiseases in patients with acute anterior uveitisrdquo ScandinavianJournal of Rheumatology vol 11 no 2 pp 121ndash123 1982
[12] A B Beckingsale J Davies J M Gibson and A RalphRosenthal ldquoAcute anterior uveitis ankylosing spondylitis backpain and HLA-B27rdquo British Journal of Ophthalmology vol 68no 10 pp 741ndash745 1984
[13] G P Thomas and M A Brown ldquoGenetics and genomics ofankylosing spondylitisrdquo Immunological Reviews vol 233 no 1pp 162ndash180 2010
[14] P C Robinson and M A Brown ldquoGenetics of ankylosingspondylitisrdquoMolecular Immunology vol 57 no 1 pp 2ndash11 2014
[15] C Lopez-Larrea K Sujirachato N K Mehra et al ldquoHLA-B27 subtypes in Asian patients with ankylosing spondylitisEvidence for new associationsrdquo Tissue Antigens vol 45 no 3pp 169ndash176 1995
[16] P C Robinson T A M Claushuis A Cortes et al ldquoGeneticdissection of acute anterior uveitis reveals similarities and dif-ferences in associations observed with ankylosing spondylitisrdquoArthritis and Rheumatology vol 67 no 1 pp 140ndash151 2015
[17] H Li S Hou H Yu et al ldquoAssociation of genetic variationsin TNFSF15 with acute anterior uveitis in Chinese HanrdquoInvestigative Ophthalmology and Visual Science vol 56 no 8pp 4605ndash4610 2015
[18] Q Xiang L Chen J Fang et al ldquoTNF receptor-associated factor5 gene confers genetic predisposition to acute anterior uveitisand pediatric uveitisrdquo Arthritis Research and Therapy vol 15no 5 article R113 2013
[19] H Yu Y Liu L Zhang et al ldquoFoxO1 gene confers genetic predis-position to acute anterior uveitis with ankylosing spondylitisrdquoInvestigative Ophthalmology amp Visual Science vol 55 no 12 pp7970ndash7974 2014
[20] T M Martin L Bye N Modi et al ldquoGenotype analysis ofpolymorphisms in autoimmune susceptibility genes CTLA-4and PTPN22 in an Acute anterior uveitis cohortrdquo MolecularVision vol 15 pp 208ndash212 2009
[21] Q Zhang J Qi S Hou et al ldquoA functional variant of PTPN22confers risk for Vogt-Koyanagi-Harada syndrome but not forankylosing spondylitisrdquo PLoS ONE vol 9 no 5 article e969432014
[22] S-S Zhao J-WHu JWang X-J Lou and L-L Zhou ldquoInversecorrelation between CD4+CD25 highCD127lowminus regulatory T-cells and serum immunoglobulin A in patients with new-onset ankylosing spondylitisrdquo Journal of International MedicalResearch vol 39 no 5 pp 1968ndash1974 2011
[23] W Zou Z Wu X Xiang S Sun and J Zhang ldquoThe expressionand significance of T helper cell subsets and regulatory Tcells CD4+CD25+ in peripheral blood of patients with humanleukocyte antigen B27-positive acute anterior uveitisrdquo GraefersquosArchive for Clinical and Experimental Ophthalmology vol 252no 4 pp 665ndash672 2014
BioMed Research International 9
[24] C Wang Q Liao Y Hu and D Zhong ldquoT lymphocyte subsetimbalances in patients contribute to ankylosing spondylitisrdquoExperimental and Therapeutic Medicine vol 9 no 1 pp 250ndash256 2015
[25] A Tedgui and Z Mallat ldquoCytokines in atherosclerosispathogenic and regulatory pathwaysrdquoPhysiological Reviews vol86 no 2 pp 515ndash581 2006
[26] P J Murray ldquoThe JAK-STAT signaling pathway Input andoutput integrationrdquo Journal of Immunology vol 178 no 5 pp2623ndash2629 2007
[27] A N Mathur H-C Chang D G Zisoulis et al ldquoStat3 andStat4 direct development of IL-17-secreting Th cellsrdquo Journal ofImmunology vol 178 no 8 pp 4901ndash4907 2007
[28] G K Hansson ldquoRegulation of immune mechanisms inatherosclerosisrdquo Annals of the New York Academy of Sciencesvol 947 pp 157ndash165 2001
[29] T Shea-Donohue A Fasano A Smith and A Zhao ldquoEntericpathogens and gut function role of cytokines and STATsrdquo GutMicrobes vol 1 no 5 pp 316ndash324 2010
[30] T Taniguchi K Ogasawara A Takaoka and N Tanaka ldquoIRFfamily of transcription factors as regulators of host defenserdquoAnnual Review of Immunology vol 19 pp 623ndash655 2001
[31] M Lohoff D Ferrick H-W Mittrucker et al ldquoInterferonregulatory factor-1 is required for a T helper 1 immune responsein vivordquo Immunity vol 6 no 6 pp 681ndash689 1997
[32] S Taki T Sato K Ogasawara et al ldquoMultistage regulation ofTh1-type immune responses by the transcription factor IRF-1rdquoImmunity vol 6 no 6 pp 673ndash679 1997
[33] C Bogdan ldquoNitric oxide and the immune responserdquo NatureImmunology vol 2 no 10 pp 907ndash916 2001
[34] S Brahmachari and K Pahan ldquoMyelin basic protein primingreduces the expression of Foxp3 in T cells via nitric oxiderdquoTheJournal of Immunology vol 184 no 4 pp 1799ndash1809 2010
[35] S-W Lee H Choi S-Y Eun S Fukuyama and M CroftldquoNitric oxide modulates TGF-120573-directive signals to suppressFoxp3 + regulatory T cell differentiation and potentiate Th1developmentrdquo Journal of Immunology vol 186 no 12 pp 6972ndash6980 2011
[36] D A Jabs R B Nussenblatt J T Rosenbaum and Standard-ization of Uveitis Nomenclature Working G ldquoStandardizationof uveitis nomenclature for reporting clinical data Resultsof the First International Workshoprdquo American Journal ofOphthalmology vol 140 pp 509ndash516 2005
[37] S Van Der Linden H A Valkenburg and A Cats ldquoEvaluationof diagnostic criteria for ankylosing spondylitis A proposal formodification of the New York criteriardquo Arthritis and Rheuma-tism vol 27 no 4 pp 361ndash368 1984
[38] D Ellinghaus E Ellinghaus R P Nair et al ldquoCombined anal-ysis of genome-wide association studies for Crohn disease andpsoriasis identifies seven shared susceptibility locirdquo AmericanJournal of Human Genetics vol 90 no 4 pp 636ndash647 2012
[39] G Fortunato G Calcagno V Bresciamorra et al ldquoMultiplesclerosis and hepatitis C virus infection are associated withsingle nucleotide polymorphisms in interferon pathway genesrdquoJournal of Interferon and Cytokine Research vol 28 no 3 pp141ndash152 2008
[40] W-H Hahn J-S Suh S H Cho B-S Cho and S-D KimldquoPolymorphisms of signal transducers and activators of tran-scription 1 and 4 (STAT1 and STAT4) contribute to progressionof childhood IgA nephropathyrdquo Cytokine vol 50 no 1 pp 69ndash74 2010
[41] S Hou J Qi Q Zhang et al ldquoGenetic variants in the JAK1 geneconfer higher risk of Behcetrsquos disease with ocular involvementin Han ChineserdquoHumanGenetics vol 132 no 9 pp 1049ndash10582013
[42] K Hu S Hou F Li Q Xiang A Kijlstra and P YangldquoJAK1 but not JAK2 and STAT3 confers susceptibility toVogt-Koyanagi-Harada (VKH) syndrome in a Han Chinesepopulationrdquo Investigative Ophthalmology and Visual Sciencevol 54 no 5 pp 3360ndash3365 2013
[43] A Cortes J Hadler J P Pointon et al ldquoIdentification ofmultiple risk variants for ankylosing spondylitis through high-density genotyping of immune-related locirdquo Nature Geneticsvol 45 no 7 pp 730ndash738 2013
[44] P A Shaiq P E Stuart A Latif et al ldquoGenetic associations ofpsoriasis in a Pakistani populationrdquo British Journal of Derma-tology vol 169 no 2 pp 406ndash411 2013
[45] S-K Yang Y Jung H Kim M Hong B D Ye and KSong ldquoAssociation of FCGR2A JAK2 or HNF4A variants withulcerative colitis in Koreansrdquo Digestive and Liver Disease vol43 no 11 pp 856ndash861 2011
[46] M Garcıa-Bermudez R Lopez-Mejıas F Genre et al ldquoLackof association between JAK3 gene polymorphisms and cardio-vascular disease in Spanish patients with rheumatoid arthritisrdquoBioMed Research International vol 2015 Article ID 318364 11pages 2015
[47] R Lopez-Mejıas M Garcıa-Bermudez C Gonzalez-Juanateyet al ldquoNFKB1-94ATTG insdel polymorphism (rs28362491) isassociated with cardiovascular disease in patients with rheuma-toid arthritisrdquo Atherosclerosis vol 224 no 2 pp 426ndash429 2012
[48] H J Park J W Kim B-S Cho and J-H Chung ldquoAssociationof BH3 interacting domain death agonist (BID) gene polymor-phisms with proteinuria of immunoglobulin A nephropathyrdquoScandinavian Journal of Clinical and Laboratory Investigationvol 74 no 4 pp 329ndash335 2014
Submit your manuscripts athttpwwwhindawicom
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Anatomy Research International
PeptidesInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporation httpwwwhindawicom
International Journal of
Volume 2014
Zoology
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Molecular Biology International
GenomicsInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
BioinformaticsAdvances in
Marine BiologyJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Signal TransductionJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
BioMed Research International
Evolutionary BiologyInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Biochemistry Research International
ArchaeaHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Genetics Research International
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Advances in
Virolog y
Hindawi Publishing Corporationhttpwwwhindawicom
Nucleic AcidsJournal of
Volume 2014
Stem CellsInternational
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
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Enzyme Research
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
International Journal of
Microbiology
BioMed Research International 5
Table 4 Allele and genotype frequencies of SNPs in patients with AAU versus control subjects stratified by HLA-B27 status
Gene SNP Allele and genotype AAUHLA-B27 Control 119875 Pc OR (95 CI)
JAK1
rs310241
C 324 (310) 392 (275) 006 NS 118 (100ndash141)CC 41 (79) 45 (63) 029 NS 127 (082ndash196)CT 242 (464) 302 (424) 016 NS 118 (094ndash148)TT 239 (457) 366 (513) 005 NS 080 (064ndash100)
rs2780815
G 878 (862) 1250 (875) 035 NS 089 (070ndash113)GG 381 (748) 548 (767) 044 NS 090 (069ndash118)GT 116 (228) 154 (216) 061 NS 107 (082ndash141)TT 12 (24) 12 (17) 040 NS 141 (063ndash317)
JAK2
rs10758669
A 705 (659) 980 (690) 010 NS 087 (073ndash103)AA 220 (411) 346 (487) 008 NS 074 (059ndash092)AC 265 (495) 288 (406) 162 times 10minus3 0053 144 (115ndash180)CC 50 (94) 76 (107) 043 NS 086 (059ndash125)
rs10975003
C 251 (245) 272 (212) 005 NS 121 (100ndash147)CC 26 (51) 28 (44) 056 NS 118 (068ndash203)CT 199 (388) 216 (335) 006 NS 126 (099ndash160)TT 287 (561) 399 (621) 003 NS 077 (061ndash098)
STAT1
rs1547550
C 937 (872) 1254 (878) 067 NS 095 (075ndash121)CC 407 (758) 555 (777) 042 NS 090 (069ndash117)CG 123 (229) 144 (202) 024 NS 118 (090ndash154)GG 7 (13) 15 (21) 029 NS 062 (025ndash152)
rs2066802
C 190 (193) 288 (207) 039 NS 091 (074ndash112)CC 15 (30) 27 (39) 044 NS 078 (041ndash148)CT 160 (325) 234 (337) 066 NS 095 (074ndash121)TT 318 (645) 434 (624) 047 NS 109 (086ndash139)
rs6718902
C 584 (565) 778 (545) 033 NS 108 (092ndash127)CC 163 (315) 209 (293) 040 NS 111 (087ndash142)CT 258 (499) 360 (504) 086 NS 098 (078ndash123)TT 96 (186) 145 (203) 045 NS 090 (067ndash119)
rs10199181
A 270 (272) 397 (278) 075 NS 097 (081ndash116)AA 43 (87) 55 (77) 055 NS 114 (075ndash173)AT 184 (371) 287 (402) 028 NS 088 (069ndash111)TT 269 (542) 372 (521) 047 NS 109 (087ndash137)
IRF1 rs2070721
A 356 (357) 487 (341) 040 NS 108(091ndash127)AA 61 (122) 74 (104) 030 NS 121 (084ndash173)AC 234 (470) 339 (474) 087 NS 098 (078ndash123)CC 203 (408) 301 (422) 063 NS 094 (075ndash119)
NOS2
rs2297518
A 194 (181) 209 (162) 021 NS 115 (093ndash142)AA 12 (23) 12 (19) 064 NS 121 (054ndash272)AG 170 (317) 185 (286) 024 NS 116 (090ndash149)GG 354 (660) 450 (695) 020 NS 085 (067ndash109)
rs4795067
A 797 (743) 1008 (761) 035 NS 092 (076ndash110)AA 293 (546) 382 (577) 033 NS 089 (071ndash123)AG 211 (394) 244 (369) 037 NS 111 (088ndash141)GG 32 (60) 36 (54) 069 NS 110 (068ndash180)
OR = odds ratio 95 CI = 95 confidence intervalPc = 119875 value adjusted by Bonferroni correction
6 BioMed Research InternationalTa
ble5Alleleandgeno
type
frequ
encies
ofSN
Psin
patientsw
ithAAU
versus
controlsub
jectsstratified
byASandHLA
-B27
status
Gene
SNP
Alleleandgeno
type
AAU+AS+
HLA
-B27
AAU+ASminus
HLA
-B27
Con
trol
119875(A
S+)
Pc(A
S+)
OR(95
CI)119875(A
Sminus)
Pc(A
Sminus)
OR(95
CI)
JAK1
rs310241
C205(316
)
119(299
)
392(275
)
005
NS
122(100ndash
149)
032
NS
113(088ndash14
5)CC
30(93)
11(55)
45(63)
009
NS
152(094ndash
245)
070
NS
087
(044ndash
172)
CT145(447
)97
(487
)302(424
)047
NS
110(085ndash14
4)010
NS
131(095ndash179)
TT149(460
)90
(457
)366(513
)
011
NS
081
(062ndash10
5)014
NS
079
(058ndash10
8)
rs2780815
G543(843
)335(896
)
1250
(875
)
005
NS
077
(059ndash
100)
028
NS
122(085ndash17
7)GG
230(714
)
151(807)
548(767
)007
NS
076
(056ndash
102)
024
NS
127(085ndash19
0)GT
83(258
)33
(176
)
154(216
)
014
NS
126(093ndash17
2)023
NS
078
(051ndash118)
TT9(28)
3(17)
12(17)
024
NS
168(070ndash
403)
094
NS
095
(027ndash342)
JAK2
rs10758669
A44
9(662
)256(653
)980(690
)
020
NS
088
(072ndash17
0)016
NS
085
(067ndash10
7)AA
139(410
)
81(413
)
346(487
)002
NS
073
(056ndash
095)
007
NS
074
(054ndash
102)
AC171(504)
94(480
)288(406
)256times10minus3
0084
149(115
ndash194)
006
NS
135(098ndash18
6)CC
29(86)
21(107
)76
(107
)028
NS
078
(050ndash
122)
100
NS
100(060ndash
167)
rs10975003
C165(257
)86
(225
)272(212
)
003
NS
129(103ndash16
1)057
NS
108(082ndash14
3)CC
18(56)
8(42)
28(44)
039
NS
131(071ndash240)
092
NS
096
(043ndash214)
CT129(402
)70
(366
)216(335
)004
NS
133(101ndash17
5)044
NS
114(082ndash16
0)TT
174(542
)113(592
)
399(621
)001
NS
072
(055ndash095)
047
NS
089
(064ndash
123)
STAT
1
rs1547550
C608(881
)329(857
)1254
(878
)
084
NS
103(078ndash13
6)026
NS
083
(060ndash
115)
CC264(765
)143(745
)555(777
)
066
NS
093
(069ndash
127)
034
NS
084
(058ndash12
1)CG
80(232
)43
(224
)144(202
)026
NS
120(088ndash16
3)050
NS
114(078ndash16
8)GG
1(03
)6(31
)15
(21
)002
NS
014
(002ndash103)
040
NS
150(058ndash393)
rs2066
802
C130(207
)60
(168
)288(207
)099
NS
100(079ndash
126)
009
NS
077
(057ndash10
5)CC
11(35)
4(22)
27(39)
077
NS
090
(044ndash
183)
029
NS
057
(020ndash
164)
CT108(344
)52
(291
)
234(337
)082
NS
103(078ndash13
7)024
NS
081
(056ndash
116)
TT195(621
)123(687
)434(624
)092
NS
099
(075ndash13
0)012
NS
132(093ndash18
8)
rs6718902
C368(577
)
216(545
)778(545
)018
NS
114(094ndash
138)
098
NS
100(080ndash
125)
CC103(323
)60
(303
)209(293
)
033
NS
115(087ndash15
3)078
NS
105(075ndash14
8)CT
162(508
)96
(485
)360(504
)091
NS
102(078ndash13
2)063
NS
093
(068ndash12
7)TT
54(169
)42
(212
)
145(203
)020
NS
080
(057ndash113)
078
NS
106(072ndash15
6)
rs10199181
A170(269
)100(278
)
397(278
)
067
NS
096
(077ndash118)
099
NS
100(077ndash12
9)AA
25(79
)18
(100
)55
(77
)091
NS
103(063ndash16
8)032
NS
133(076
ndash233
)AT
120(380
)64
(356
)287(402
)050
NS
091
(069ndash
120)
026
NS
082
(058ndash115)
TT171(541
)98
(544
)372(521
)055
NS
108(083ndash14
1)057
NS
110(079ndash
153)
IRF1
rs2070721
A221(349)
135(373
)
487(341
)073
NS
103(085ndash12
6)025
NS
115(090-14
6)AA
37(117
)
24(133
)74
(104
)053
NS
114(075ndash174)
026
NS
132(081ndash216)
AC147(464
)87
(481
)339(474
)
074
NS
096
(073ndash12
5)089
NS
102(074
ndash142)
CC133(420
)70
(386
)301(422)
095
NS
099
(076
ndash130)
040
NS
097
(062ndash12
1)
NOS2
rs2297518
A130(19
0)
64(165
)209(162
)011
NS
122(096ndash
155)
087
NS
103(076
ndash139
)AA
7(20)
5(26)
12(19)
083
NS
111(043ndash284)
053
NS
140(049ndash
402)
AG116(339
)54
(278
)
185(286
)008
NS
128(097ndash17
0)084
NS
096
(067ndash13
8)GG
219(641
)135(696
)
450(695
)
007
NS
078
(059ndash
103)
099
NS
100(071ndash14
2)
rs4795067
A512(736
)295(745
)1008
(761
)020
NS
087
(071ndash10
8)061
NS
092
(071ndash119)
AA
184(529
)109(551
)382(577
)
014
NS
082
(063ndash10
7)051
NS
090
(065ndash12
4)AG
144(414
)
77(388
)244(369
)016
NS
121(093ndash158)
060
NS
109(079ndash
151)
GG
20(57)
12(61
)36
(54)
084
NS
106(060ndash
186)
074
NS
112(057ndash220)
OR=od
dsratio
95
CI=95
confi
denceinterval
Pc=119875valuea
djustedby
Bonferroni
correctio
n
BioMed Research International 7
differences in the genotype and allele frequencies of the other10 SNPs were observed between the HLA-B27-positive AS-positiveAAUpatients and healthy controls (119875Bonferroni gt 005Table 5) In addition there were no significant differencesin the genotype and allele frequencies of the tested SNPsbetween the AS-positive AAU patients and control subjects(119875Bonferroni gt 005 Supplementary Table S3)
33 Logistic Regression Analysis of SNPs in AAU We furtherinvestigated the SNPs rs10758669 and rs10975003 using addi-tive codominant dominant and recessive genetic modelsusing a multivariate logistic regression model adjusted forage and gender We observed that the frequency of the CAgenotype of rs10758669 was significantly higher in AAUpatients which suggests that patients with the rs10758669CA genotype have increased susceptibility to AAU (468versus 406 OR = 128 119875 = 002 SupplementaryTable S4) An increased frequency of the AC genotype ofrs10758669 was also observed in HLA-B27-positive AAUpatients and AS-positive AAU patients compared to thehealthy controls (495 versus 406 OR = 143 119875 =350 times 10minus3 and 492 versus 406 OR = 136 119875 = 002Supplementary Table S4) For SNP rs10975003 the frequencyof the heterozygous CT genotype was significantly higherin female AAU patients compared to the healthy controls(404 versus 336 OR = 157 119875 = 001 SupplementaryTable S5) A similar result was observed when we combinedCT and CC to construct a dominant model (443 versus380 OR = 157 119875 = 910times 10minus3 Supplementary Table S5)However none of the observed associations for the two SNPsretained statistical significance after Bonferroni correction(119875Bonferroni gt 005) Furthermore no significance associationswere found between the other SNPs and AAU even afterstratification by gender AS and HLA-B27 status (data notshown)
4 Discussion
In this study we first investigated whether genetic poly-morphisms of JAK-STAT signaling pathway genes includingJAK1 JAK2 STAT1 IRF1 and NOS2 confer susceptibilityto AAU with or without AS in a Chinese Han populationOur results suggest that none of these SNPs exhibit statisti-cally different frequencies of genotypes and alleles betweenhealthy controls and AAU patients However we observeda boundary significant association for two SNPs (rs10975003and rs10758669) of JAK2 by stratification analysis by genderand HLA-B27 status
We highlighted two issues at the time of study designto obtain unbiased association results First we followedstrict criteria for the diagnosis of AAU patients Patients withAAU were diagnosed as previously described by Jabs et al[36] and patients with AS were diagnosed with the modifiedNew York Criteria [37] In addition the AAU patients andhealthy controls were strictly matched by ethnicity and ageto avoid a possible influence of population stratificationFurthermore we only enrolled controls who had detailedhistories and physical examinations and excluded controlswith any autoimmune or immune-related diseases Finally
20 of the samples were randomly chosen and analyzed bydirect sequencing and the results of different genotypingmethods were consistent
AAU is defined as inflammation confined to the anteriorsegment of the eye that involves the iris and anterior part ofthe ciliary body The HLA-B27 is considered to be stronglyassociated with both AAU and AS [5 8 10 14] Recentgenetic studies have revealed that five candidate genes inthe JAK-STAT signaling pathway were considered geneticpredisposing factors for different autoimmune-mediated dis-eases [38ndash45] SNPs (rs10758669 and 10975003) in JAK2 areconsidered susceptibility factors for Crohnrsquos disease (CD) inthe German population and ulcerative colitis (UC) in theKorean population [38 45] One SNP (rs2070721) in IRF1 andSNPs (rs2297518 and rs4795067) in NOS2 are also associatedwith autoimmune diseases such as multiple sclerosis (MS)in Italy AS in Europe and psoriasis in Pakistan [39 43 44]Four SNPs (rs6718902 rs10199181 rs2066802 and rs1547550)in the STAT1 gene were observed to be associated with MSin Italy and IgA nephropathy (IgAN) in Korea [39 40] Inaddition an association was found between SNPs (rs2780815rs310241 rs3790532 rs310230 and rs310236) in JAK1 andBehcet disease (BD) as well as Vogt-Koyanagi-Harada (VKH)syndrome [41 42] two other common uveitis entities inChina There has been no report of associations betweenJAK-STAT signaling pathway genes and AAU and thus weperformed this case-control study to detect whether the fivecandidate genes were associated with AAU in a Chinese Hanpopulation Our results showed that there were no significantassociations between the genetic polymorphisms of the fivecandidate genes in the JAK-STAT signaling pathway andAAUThese results are not consistent with those observed forother autoimmune diseases reported in German Europeanand some other Asian populations [38 43 45] This discrep-ancy may be attributable to differences in the etiology andpathogenesis of AAU compared with BD VKH syndromeand autoimmune-mediated diseases
Consistent with our results a recent study also reportedno significant association of JAK-STAT signaling pathwaygene polymorphisms with rheumatoid arthritis stratified bythe presenceabsence of cardiovascular disease [46] Con-versely an influence of NFKB1 signaling pathway poly-morphisms on the development of cardiovascular events inpatients with rheumatoid arthritis has been observed [47]Furthermore NFKB1 signaling pathway polymorphismshave been described to play a critical role in the developmentof many autoimmune and inflammatory diseases and thusthe evaluation of the potential relationships between NFKB1polymorphisms and the development of AAU could be apromising research line for the future
There were several limitations of our studyWe had a lim-ited sample size to detect SNPs with weak effects while con-sidering multiple corrections Even for SNPs rs10758669 andrs10975003 we only had 700 power using a genetic powercalculator [48] In addition our samples were restricted to theHan Chinese population and all patients were enrolled fromthe ophthalmology department Further studies with a largersample size and other ethnic populations as well as patientsenrolled from multiple sources are warranted to confirm our
8 BioMed Research International
findings Additionally we only focused on eleven SNPs in theJAK-STAT pathway and it is possible that other unknownSNPs might be associated with AAU risk
In conclusion this study reveals that genetic polymor-phisms of JAK-STAT pathway genes including JAK1 JAK2STAT1 IRF1 andNOS2 may not be involved in susceptibilityto AAU risk in the Han Chinese population
Competing Interests
The authors declare that there is no conflict of interestsregarding the publication of this paper
Authorsrsquo Contributions
Ling Cheng and Hongsong Yu contributed equally to thiswork
Acknowledgments
The authors would like to thank Professor Peizeng Yang forhis great help in our study They are also grateful to alldonors enrolled in this study This work was supported bythe National Natural Science Foundation Project (8120067881670844) Fundamental and Advanced Research Programof Chongqing (cstc2015jcyjA10022) Science and TechnologyProject of Chongqing Municipal Education Commission(KJ1500236) Scientific Research Program of Science andTechnology Commission of Yuzhong District of Chongqing(20150102) and National Key Clinical Specialties Construc-tion Program of China
References
[1] D-K Hwang Y-J Chou C-Y Pu and P Chou ldquoEpidemi-ology of uveitis among the Chinese population in Taiwan APopulation-Based Studyrdquo Ophthalmology vol 119 no 11 pp2371ndash2376 2012
[2] K Nakao and N Ohba ldquoPrevalence of endogenous uveitis inKagoshima Prefecture Southwest JapanrdquoNipponGankaGakkaiZasshi vol 100 no 2 pp 150ndash155 1996
[3] D C Gritz and I GWong ldquoIncidence and prevalence of uveitisin Northern California the Northern California Epidemiologyof Uveitis Studyrdquo Ophthalmology vol 111 no 3 pp 491ndash5002004
[4] R B Nussenblatt and I Gery ldquoExperimental autoimmuneuveitis and its relationship to clinical ocular inflammatorydiseaserdquo Journal of Autoimmunity vol 9 no 5 pp 575ndash5851996
[5] P Yang Z Zhang H Zhou et al ldquoClinical patterns andcharacteristics of uveitis in a tertiary center for uveitis in ChinardquoCurrent Eye Research vol 30 no 11 pp 943ndash948 2005
[6] J T Gran and J F Skomsvoll ldquoThe outcome of ankylosingspondylitis a study of 100 patientsrdquo British Journal of Rheuma-tology vol 36 no 7 pp 766ndash771 1997
[7] L P Robertson andM J Davis ldquoA longitudinal study of diseaseactivity and functional status in a hospital cohort of patientswith ankylosing spondylitisrdquo Rheumatology vol 43 no 12 pp1565ndash1568 2004
[8] J H Chang P J McCluskey and D Wakefield ldquoAcute anterioruveitis and HLA-B27rdquo Survey of Ophthalmology vol 50 no 4pp 364ndash388 2005
[9] M Huhtinen and A Karma ldquoHLA-B27 typing in the categori-sation of uveitis in a HLA-B27 rich populationrdquo British Journalof Ophthalmology vol 84 no 4 pp 413ndash416 2000
[10] S Torres S Borges and A Artiles ldquoHLA-B27 and clinicalfeatures of acute anterior uveitis in Cubardquo Ocular Immunologyand Inflammation vol 21 no 2 pp 119ndash123 2013
[11] R Saari R Lahti K M Saari et al ldquoFrequency of rheumaticdiseases in patients with acute anterior uveitisrdquo ScandinavianJournal of Rheumatology vol 11 no 2 pp 121ndash123 1982
[12] A B Beckingsale J Davies J M Gibson and A RalphRosenthal ldquoAcute anterior uveitis ankylosing spondylitis backpain and HLA-B27rdquo British Journal of Ophthalmology vol 68no 10 pp 741ndash745 1984
[13] G P Thomas and M A Brown ldquoGenetics and genomics ofankylosing spondylitisrdquo Immunological Reviews vol 233 no 1pp 162ndash180 2010
[14] P C Robinson and M A Brown ldquoGenetics of ankylosingspondylitisrdquoMolecular Immunology vol 57 no 1 pp 2ndash11 2014
[15] C Lopez-Larrea K Sujirachato N K Mehra et al ldquoHLA-B27 subtypes in Asian patients with ankylosing spondylitisEvidence for new associationsrdquo Tissue Antigens vol 45 no 3pp 169ndash176 1995
[16] P C Robinson T A M Claushuis A Cortes et al ldquoGeneticdissection of acute anterior uveitis reveals similarities and dif-ferences in associations observed with ankylosing spondylitisrdquoArthritis and Rheumatology vol 67 no 1 pp 140ndash151 2015
[17] H Li S Hou H Yu et al ldquoAssociation of genetic variationsin TNFSF15 with acute anterior uveitis in Chinese HanrdquoInvestigative Ophthalmology and Visual Science vol 56 no 8pp 4605ndash4610 2015
[18] Q Xiang L Chen J Fang et al ldquoTNF receptor-associated factor5 gene confers genetic predisposition to acute anterior uveitisand pediatric uveitisrdquo Arthritis Research and Therapy vol 15no 5 article R113 2013
[19] H Yu Y Liu L Zhang et al ldquoFoxO1 gene confers genetic predis-position to acute anterior uveitis with ankylosing spondylitisrdquoInvestigative Ophthalmology amp Visual Science vol 55 no 12 pp7970ndash7974 2014
[20] T M Martin L Bye N Modi et al ldquoGenotype analysis ofpolymorphisms in autoimmune susceptibility genes CTLA-4and PTPN22 in an Acute anterior uveitis cohortrdquo MolecularVision vol 15 pp 208ndash212 2009
[21] Q Zhang J Qi S Hou et al ldquoA functional variant of PTPN22confers risk for Vogt-Koyanagi-Harada syndrome but not forankylosing spondylitisrdquo PLoS ONE vol 9 no 5 article e969432014
[22] S-S Zhao J-WHu JWang X-J Lou and L-L Zhou ldquoInversecorrelation between CD4+CD25 highCD127lowminus regulatory T-cells and serum immunoglobulin A in patients with new-onset ankylosing spondylitisrdquo Journal of International MedicalResearch vol 39 no 5 pp 1968ndash1974 2011
[23] W Zou Z Wu X Xiang S Sun and J Zhang ldquoThe expressionand significance of T helper cell subsets and regulatory Tcells CD4+CD25+ in peripheral blood of patients with humanleukocyte antigen B27-positive acute anterior uveitisrdquo GraefersquosArchive for Clinical and Experimental Ophthalmology vol 252no 4 pp 665ndash672 2014
BioMed Research International 9
[24] C Wang Q Liao Y Hu and D Zhong ldquoT lymphocyte subsetimbalances in patients contribute to ankylosing spondylitisrdquoExperimental and Therapeutic Medicine vol 9 no 1 pp 250ndash256 2015
[25] A Tedgui and Z Mallat ldquoCytokines in atherosclerosispathogenic and regulatory pathwaysrdquoPhysiological Reviews vol86 no 2 pp 515ndash581 2006
[26] P J Murray ldquoThe JAK-STAT signaling pathway Input andoutput integrationrdquo Journal of Immunology vol 178 no 5 pp2623ndash2629 2007
[27] A N Mathur H-C Chang D G Zisoulis et al ldquoStat3 andStat4 direct development of IL-17-secreting Th cellsrdquo Journal ofImmunology vol 178 no 8 pp 4901ndash4907 2007
[28] G K Hansson ldquoRegulation of immune mechanisms inatherosclerosisrdquo Annals of the New York Academy of Sciencesvol 947 pp 157ndash165 2001
[29] T Shea-Donohue A Fasano A Smith and A Zhao ldquoEntericpathogens and gut function role of cytokines and STATsrdquo GutMicrobes vol 1 no 5 pp 316ndash324 2010
[30] T Taniguchi K Ogasawara A Takaoka and N Tanaka ldquoIRFfamily of transcription factors as regulators of host defenserdquoAnnual Review of Immunology vol 19 pp 623ndash655 2001
[31] M Lohoff D Ferrick H-W Mittrucker et al ldquoInterferonregulatory factor-1 is required for a T helper 1 immune responsein vivordquo Immunity vol 6 no 6 pp 681ndash689 1997
[32] S Taki T Sato K Ogasawara et al ldquoMultistage regulation ofTh1-type immune responses by the transcription factor IRF-1rdquoImmunity vol 6 no 6 pp 673ndash679 1997
[33] C Bogdan ldquoNitric oxide and the immune responserdquo NatureImmunology vol 2 no 10 pp 907ndash916 2001
[34] S Brahmachari and K Pahan ldquoMyelin basic protein primingreduces the expression of Foxp3 in T cells via nitric oxiderdquoTheJournal of Immunology vol 184 no 4 pp 1799ndash1809 2010
[35] S-W Lee H Choi S-Y Eun S Fukuyama and M CroftldquoNitric oxide modulates TGF-120573-directive signals to suppressFoxp3 + regulatory T cell differentiation and potentiate Th1developmentrdquo Journal of Immunology vol 186 no 12 pp 6972ndash6980 2011
[36] D A Jabs R B Nussenblatt J T Rosenbaum and Standard-ization of Uveitis Nomenclature Working G ldquoStandardizationof uveitis nomenclature for reporting clinical data Resultsof the First International Workshoprdquo American Journal ofOphthalmology vol 140 pp 509ndash516 2005
[37] S Van Der Linden H A Valkenburg and A Cats ldquoEvaluationof diagnostic criteria for ankylosing spondylitis A proposal formodification of the New York criteriardquo Arthritis and Rheuma-tism vol 27 no 4 pp 361ndash368 1984
[38] D Ellinghaus E Ellinghaus R P Nair et al ldquoCombined anal-ysis of genome-wide association studies for Crohn disease andpsoriasis identifies seven shared susceptibility locirdquo AmericanJournal of Human Genetics vol 90 no 4 pp 636ndash647 2012
[39] G Fortunato G Calcagno V Bresciamorra et al ldquoMultiplesclerosis and hepatitis C virus infection are associated withsingle nucleotide polymorphisms in interferon pathway genesrdquoJournal of Interferon and Cytokine Research vol 28 no 3 pp141ndash152 2008
[40] W-H Hahn J-S Suh S H Cho B-S Cho and S-D KimldquoPolymorphisms of signal transducers and activators of tran-scription 1 and 4 (STAT1 and STAT4) contribute to progressionof childhood IgA nephropathyrdquo Cytokine vol 50 no 1 pp 69ndash74 2010
[41] S Hou J Qi Q Zhang et al ldquoGenetic variants in the JAK1 geneconfer higher risk of Behcetrsquos disease with ocular involvementin Han ChineserdquoHumanGenetics vol 132 no 9 pp 1049ndash10582013
[42] K Hu S Hou F Li Q Xiang A Kijlstra and P YangldquoJAK1 but not JAK2 and STAT3 confers susceptibility toVogt-Koyanagi-Harada (VKH) syndrome in a Han Chinesepopulationrdquo Investigative Ophthalmology and Visual Sciencevol 54 no 5 pp 3360ndash3365 2013
[43] A Cortes J Hadler J P Pointon et al ldquoIdentification ofmultiple risk variants for ankylosing spondylitis through high-density genotyping of immune-related locirdquo Nature Geneticsvol 45 no 7 pp 730ndash738 2013
[44] P A Shaiq P E Stuart A Latif et al ldquoGenetic associations ofpsoriasis in a Pakistani populationrdquo British Journal of Derma-tology vol 169 no 2 pp 406ndash411 2013
[45] S-K Yang Y Jung H Kim M Hong B D Ye and KSong ldquoAssociation of FCGR2A JAK2 or HNF4A variants withulcerative colitis in Koreansrdquo Digestive and Liver Disease vol43 no 11 pp 856ndash861 2011
[46] M Garcıa-Bermudez R Lopez-Mejıas F Genre et al ldquoLackof association between JAK3 gene polymorphisms and cardio-vascular disease in Spanish patients with rheumatoid arthritisrdquoBioMed Research International vol 2015 Article ID 318364 11pages 2015
[47] R Lopez-Mejıas M Garcıa-Bermudez C Gonzalez-Juanateyet al ldquoNFKB1-94ATTG insdel polymorphism (rs28362491) isassociated with cardiovascular disease in patients with rheuma-toid arthritisrdquo Atherosclerosis vol 224 no 2 pp 426ndash429 2012
[48] H J Park J W Kim B-S Cho and J-H Chung ldquoAssociationof BH3 interacting domain death agonist (BID) gene polymor-phisms with proteinuria of immunoglobulin A nephropathyrdquoScandinavian Journal of Clinical and Laboratory Investigationvol 74 no 4 pp 329ndash335 2014
Submit your manuscripts athttpwwwhindawicom
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Anatomy Research International
PeptidesInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporation httpwwwhindawicom
International Journal of
Volume 2014
Zoology
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Molecular Biology International
GenomicsInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
BioinformaticsAdvances in
Marine BiologyJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Signal TransductionJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
BioMed Research International
Evolutionary BiologyInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Biochemistry Research International
ArchaeaHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Genetics Research International
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Advances in
Virolog y
Hindawi Publishing Corporationhttpwwwhindawicom
Nucleic AcidsJournal of
Volume 2014
Stem CellsInternational
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Enzyme Research
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
International Journal of
Microbiology
6 BioMed Research InternationalTa
ble5Alleleandgeno
type
frequ
encies
ofSN
Psin
patientsw
ithAAU
versus
controlsub
jectsstratified
byASandHLA
-B27
status
Gene
SNP
Alleleandgeno
type
AAU+AS+
HLA
-B27
AAU+ASminus
HLA
-B27
Con
trol
119875(A
S+)
Pc(A
S+)
OR(95
CI)119875(A
Sminus)
Pc(A
Sminus)
OR(95
CI)
JAK1
rs310241
C205(316
)
119(299
)
392(275
)
005
NS
122(100ndash
149)
032
NS
113(088ndash14
5)CC
30(93)
11(55)
45(63)
009
NS
152(094ndash
245)
070
NS
087
(044ndash
172)
CT145(447
)97
(487
)302(424
)047
NS
110(085ndash14
4)010
NS
131(095ndash179)
TT149(460
)90
(457
)366(513
)
011
NS
081
(062ndash10
5)014
NS
079
(058ndash10
8)
rs2780815
G543(843
)335(896
)
1250
(875
)
005
NS
077
(059ndash
100)
028
NS
122(085ndash17
7)GG
230(714
)
151(807)
548(767
)007
NS
076
(056ndash
102)
024
NS
127(085ndash19
0)GT
83(258
)33
(176
)
154(216
)
014
NS
126(093ndash17
2)023
NS
078
(051ndash118)
TT9(28)
3(17)
12(17)
024
NS
168(070ndash
403)
094
NS
095
(027ndash342)
JAK2
rs10758669
A44
9(662
)256(653
)980(690
)
020
NS
088
(072ndash17
0)016
NS
085
(067ndash10
7)AA
139(410
)
81(413
)
346(487
)002
NS
073
(056ndash
095)
007
NS
074
(054ndash
102)
AC171(504)
94(480
)288(406
)256times10minus3
0084
149(115
ndash194)
006
NS
135(098ndash18
6)CC
29(86)
21(107
)76
(107
)028
NS
078
(050ndash
122)
100
NS
100(060ndash
167)
rs10975003
C165(257
)86
(225
)272(212
)
003
NS
129(103ndash16
1)057
NS
108(082ndash14
3)CC
18(56)
8(42)
28(44)
039
NS
131(071ndash240)
092
NS
096
(043ndash214)
CT129(402
)70
(366
)216(335
)004
NS
133(101ndash17
5)044
NS
114(082ndash16
0)TT
174(542
)113(592
)
399(621
)001
NS
072
(055ndash095)
047
NS
089
(064ndash
123)
STAT
1
rs1547550
C608(881
)329(857
)1254
(878
)
084
NS
103(078ndash13
6)026
NS
083
(060ndash
115)
CC264(765
)143(745
)555(777
)
066
NS
093
(069ndash
127)
034
NS
084
(058ndash12
1)CG
80(232
)43
(224
)144(202
)026
NS
120(088ndash16
3)050
NS
114(078ndash16
8)GG
1(03
)6(31
)15
(21
)002
NS
014
(002ndash103)
040
NS
150(058ndash393)
rs2066
802
C130(207
)60
(168
)288(207
)099
NS
100(079ndash
126)
009
NS
077
(057ndash10
5)CC
11(35)
4(22)
27(39)
077
NS
090
(044ndash
183)
029
NS
057
(020ndash
164)
CT108(344
)52
(291
)
234(337
)082
NS
103(078ndash13
7)024
NS
081
(056ndash
116)
TT195(621
)123(687
)434(624
)092
NS
099
(075ndash13
0)012
NS
132(093ndash18
8)
rs6718902
C368(577
)
216(545
)778(545
)018
NS
114(094ndash
138)
098
NS
100(080ndash
125)
CC103(323
)60
(303
)209(293
)
033
NS
115(087ndash15
3)078
NS
105(075ndash14
8)CT
162(508
)96
(485
)360(504
)091
NS
102(078ndash13
2)063
NS
093
(068ndash12
7)TT
54(169
)42
(212
)
145(203
)020
NS
080
(057ndash113)
078
NS
106(072ndash15
6)
rs10199181
A170(269
)100(278
)
397(278
)
067
NS
096
(077ndash118)
099
NS
100(077ndash12
9)AA
25(79
)18
(100
)55
(77
)091
NS
103(063ndash16
8)032
NS
133(076
ndash233
)AT
120(380
)64
(356
)287(402
)050
NS
091
(069ndash
120)
026
NS
082
(058ndash115)
TT171(541
)98
(544
)372(521
)055
NS
108(083ndash14
1)057
NS
110(079ndash
153)
IRF1
rs2070721
A221(349)
135(373
)
487(341
)073
NS
103(085ndash12
6)025
NS
115(090-14
6)AA
37(117
)
24(133
)74
(104
)053
NS
114(075ndash174)
026
NS
132(081ndash216)
AC147(464
)87
(481
)339(474
)
074
NS
096
(073ndash12
5)089
NS
102(074
ndash142)
CC133(420
)70
(386
)301(422)
095
NS
099
(076
ndash130)
040
NS
097
(062ndash12
1)
NOS2
rs2297518
A130(19
0)
64(165
)209(162
)011
NS
122(096ndash
155)
087
NS
103(076
ndash139
)AA
7(20)
5(26)
12(19)
083
NS
111(043ndash284)
053
NS
140(049ndash
402)
AG116(339
)54
(278
)
185(286
)008
NS
128(097ndash17
0)084
NS
096
(067ndash13
8)GG
219(641
)135(696
)
450(695
)
007
NS
078
(059ndash
103)
099
NS
100(071ndash14
2)
rs4795067
A512(736
)295(745
)1008
(761
)020
NS
087
(071ndash10
8)061
NS
092
(071ndash119)
AA
184(529
)109(551
)382(577
)
014
NS
082
(063ndash10
7)051
NS
090
(065ndash12
4)AG
144(414
)
77(388
)244(369
)016
NS
121(093ndash158)
060
NS
109(079ndash
151)
GG
20(57)
12(61
)36
(54)
084
NS
106(060ndash
186)
074
NS
112(057ndash220)
OR=od
dsratio
95
CI=95
confi
denceinterval
Pc=119875valuea
djustedby
Bonferroni
correctio
n
BioMed Research International 7
differences in the genotype and allele frequencies of the other10 SNPs were observed between the HLA-B27-positive AS-positiveAAUpatients and healthy controls (119875Bonferroni gt 005Table 5) In addition there were no significant differencesin the genotype and allele frequencies of the tested SNPsbetween the AS-positive AAU patients and control subjects(119875Bonferroni gt 005 Supplementary Table S3)
33 Logistic Regression Analysis of SNPs in AAU We furtherinvestigated the SNPs rs10758669 and rs10975003 using addi-tive codominant dominant and recessive genetic modelsusing a multivariate logistic regression model adjusted forage and gender We observed that the frequency of the CAgenotype of rs10758669 was significantly higher in AAUpatients which suggests that patients with the rs10758669CA genotype have increased susceptibility to AAU (468versus 406 OR = 128 119875 = 002 SupplementaryTable S4) An increased frequency of the AC genotype ofrs10758669 was also observed in HLA-B27-positive AAUpatients and AS-positive AAU patients compared to thehealthy controls (495 versus 406 OR = 143 119875 =350 times 10minus3 and 492 versus 406 OR = 136 119875 = 002Supplementary Table S4) For SNP rs10975003 the frequencyof the heterozygous CT genotype was significantly higherin female AAU patients compared to the healthy controls(404 versus 336 OR = 157 119875 = 001 SupplementaryTable S5) A similar result was observed when we combinedCT and CC to construct a dominant model (443 versus380 OR = 157 119875 = 910times 10minus3 Supplementary Table S5)However none of the observed associations for the two SNPsretained statistical significance after Bonferroni correction(119875Bonferroni gt 005) Furthermore no significance associationswere found between the other SNPs and AAU even afterstratification by gender AS and HLA-B27 status (data notshown)
4 Discussion
In this study we first investigated whether genetic poly-morphisms of JAK-STAT signaling pathway genes includingJAK1 JAK2 STAT1 IRF1 and NOS2 confer susceptibilityto AAU with or without AS in a Chinese Han populationOur results suggest that none of these SNPs exhibit statisti-cally different frequencies of genotypes and alleles betweenhealthy controls and AAU patients However we observeda boundary significant association for two SNPs (rs10975003and rs10758669) of JAK2 by stratification analysis by genderand HLA-B27 status
We highlighted two issues at the time of study designto obtain unbiased association results First we followedstrict criteria for the diagnosis of AAU patients Patients withAAU were diagnosed as previously described by Jabs et al[36] and patients with AS were diagnosed with the modifiedNew York Criteria [37] In addition the AAU patients andhealthy controls were strictly matched by ethnicity and ageto avoid a possible influence of population stratificationFurthermore we only enrolled controls who had detailedhistories and physical examinations and excluded controlswith any autoimmune or immune-related diseases Finally
20 of the samples were randomly chosen and analyzed bydirect sequencing and the results of different genotypingmethods were consistent
AAU is defined as inflammation confined to the anteriorsegment of the eye that involves the iris and anterior part ofthe ciliary body The HLA-B27 is considered to be stronglyassociated with both AAU and AS [5 8 10 14] Recentgenetic studies have revealed that five candidate genes inthe JAK-STAT signaling pathway were considered geneticpredisposing factors for different autoimmune-mediated dis-eases [38ndash45] SNPs (rs10758669 and 10975003) in JAK2 areconsidered susceptibility factors for Crohnrsquos disease (CD) inthe German population and ulcerative colitis (UC) in theKorean population [38 45] One SNP (rs2070721) in IRF1 andSNPs (rs2297518 and rs4795067) in NOS2 are also associatedwith autoimmune diseases such as multiple sclerosis (MS)in Italy AS in Europe and psoriasis in Pakistan [39 43 44]Four SNPs (rs6718902 rs10199181 rs2066802 and rs1547550)in the STAT1 gene were observed to be associated with MSin Italy and IgA nephropathy (IgAN) in Korea [39 40] Inaddition an association was found between SNPs (rs2780815rs310241 rs3790532 rs310230 and rs310236) in JAK1 andBehcet disease (BD) as well as Vogt-Koyanagi-Harada (VKH)syndrome [41 42] two other common uveitis entities inChina There has been no report of associations betweenJAK-STAT signaling pathway genes and AAU and thus weperformed this case-control study to detect whether the fivecandidate genes were associated with AAU in a Chinese Hanpopulation Our results showed that there were no significantassociations between the genetic polymorphisms of the fivecandidate genes in the JAK-STAT signaling pathway andAAUThese results are not consistent with those observed forother autoimmune diseases reported in German Europeanand some other Asian populations [38 43 45] This discrep-ancy may be attributable to differences in the etiology andpathogenesis of AAU compared with BD VKH syndromeand autoimmune-mediated diseases
Consistent with our results a recent study also reportedno significant association of JAK-STAT signaling pathwaygene polymorphisms with rheumatoid arthritis stratified bythe presenceabsence of cardiovascular disease [46] Con-versely an influence of NFKB1 signaling pathway poly-morphisms on the development of cardiovascular events inpatients with rheumatoid arthritis has been observed [47]Furthermore NFKB1 signaling pathway polymorphismshave been described to play a critical role in the developmentof many autoimmune and inflammatory diseases and thusthe evaluation of the potential relationships between NFKB1polymorphisms and the development of AAU could be apromising research line for the future
There were several limitations of our studyWe had a lim-ited sample size to detect SNPs with weak effects while con-sidering multiple corrections Even for SNPs rs10758669 andrs10975003 we only had 700 power using a genetic powercalculator [48] In addition our samples were restricted to theHan Chinese population and all patients were enrolled fromthe ophthalmology department Further studies with a largersample size and other ethnic populations as well as patientsenrolled from multiple sources are warranted to confirm our
8 BioMed Research International
findings Additionally we only focused on eleven SNPs in theJAK-STAT pathway and it is possible that other unknownSNPs might be associated with AAU risk
In conclusion this study reveals that genetic polymor-phisms of JAK-STAT pathway genes including JAK1 JAK2STAT1 IRF1 andNOS2 may not be involved in susceptibilityto AAU risk in the Han Chinese population
Competing Interests
The authors declare that there is no conflict of interestsregarding the publication of this paper
Authorsrsquo Contributions
Ling Cheng and Hongsong Yu contributed equally to thiswork
Acknowledgments
The authors would like to thank Professor Peizeng Yang forhis great help in our study They are also grateful to alldonors enrolled in this study This work was supported bythe National Natural Science Foundation Project (8120067881670844) Fundamental and Advanced Research Programof Chongqing (cstc2015jcyjA10022) Science and TechnologyProject of Chongqing Municipal Education Commission(KJ1500236) Scientific Research Program of Science andTechnology Commission of Yuzhong District of Chongqing(20150102) and National Key Clinical Specialties Construc-tion Program of China
References
[1] D-K Hwang Y-J Chou C-Y Pu and P Chou ldquoEpidemi-ology of uveitis among the Chinese population in Taiwan APopulation-Based Studyrdquo Ophthalmology vol 119 no 11 pp2371ndash2376 2012
[2] K Nakao and N Ohba ldquoPrevalence of endogenous uveitis inKagoshima Prefecture Southwest JapanrdquoNipponGankaGakkaiZasshi vol 100 no 2 pp 150ndash155 1996
[3] D C Gritz and I GWong ldquoIncidence and prevalence of uveitisin Northern California the Northern California Epidemiologyof Uveitis Studyrdquo Ophthalmology vol 111 no 3 pp 491ndash5002004
[4] R B Nussenblatt and I Gery ldquoExperimental autoimmuneuveitis and its relationship to clinical ocular inflammatorydiseaserdquo Journal of Autoimmunity vol 9 no 5 pp 575ndash5851996
[5] P Yang Z Zhang H Zhou et al ldquoClinical patterns andcharacteristics of uveitis in a tertiary center for uveitis in ChinardquoCurrent Eye Research vol 30 no 11 pp 943ndash948 2005
[6] J T Gran and J F Skomsvoll ldquoThe outcome of ankylosingspondylitis a study of 100 patientsrdquo British Journal of Rheuma-tology vol 36 no 7 pp 766ndash771 1997
[7] L P Robertson andM J Davis ldquoA longitudinal study of diseaseactivity and functional status in a hospital cohort of patientswith ankylosing spondylitisrdquo Rheumatology vol 43 no 12 pp1565ndash1568 2004
[8] J H Chang P J McCluskey and D Wakefield ldquoAcute anterioruveitis and HLA-B27rdquo Survey of Ophthalmology vol 50 no 4pp 364ndash388 2005
[9] M Huhtinen and A Karma ldquoHLA-B27 typing in the categori-sation of uveitis in a HLA-B27 rich populationrdquo British Journalof Ophthalmology vol 84 no 4 pp 413ndash416 2000
[10] S Torres S Borges and A Artiles ldquoHLA-B27 and clinicalfeatures of acute anterior uveitis in Cubardquo Ocular Immunologyand Inflammation vol 21 no 2 pp 119ndash123 2013
[11] R Saari R Lahti K M Saari et al ldquoFrequency of rheumaticdiseases in patients with acute anterior uveitisrdquo ScandinavianJournal of Rheumatology vol 11 no 2 pp 121ndash123 1982
[12] A B Beckingsale J Davies J M Gibson and A RalphRosenthal ldquoAcute anterior uveitis ankylosing spondylitis backpain and HLA-B27rdquo British Journal of Ophthalmology vol 68no 10 pp 741ndash745 1984
[13] G P Thomas and M A Brown ldquoGenetics and genomics ofankylosing spondylitisrdquo Immunological Reviews vol 233 no 1pp 162ndash180 2010
[14] P C Robinson and M A Brown ldquoGenetics of ankylosingspondylitisrdquoMolecular Immunology vol 57 no 1 pp 2ndash11 2014
[15] C Lopez-Larrea K Sujirachato N K Mehra et al ldquoHLA-B27 subtypes in Asian patients with ankylosing spondylitisEvidence for new associationsrdquo Tissue Antigens vol 45 no 3pp 169ndash176 1995
[16] P C Robinson T A M Claushuis A Cortes et al ldquoGeneticdissection of acute anterior uveitis reveals similarities and dif-ferences in associations observed with ankylosing spondylitisrdquoArthritis and Rheumatology vol 67 no 1 pp 140ndash151 2015
[17] H Li S Hou H Yu et al ldquoAssociation of genetic variationsin TNFSF15 with acute anterior uveitis in Chinese HanrdquoInvestigative Ophthalmology and Visual Science vol 56 no 8pp 4605ndash4610 2015
[18] Q Xiang L Chen J Fang et al ldquoTNF receptor-associated factor5 gene confers genetic predisposition to acute anterior uveitisand pediatric uveitisrdquo Arthritis Research and Therapy vol 15no 5 article R113 2013
[19] H Yu Y Liu L Zhang et al ldquoFoxO1 gene confers genetic predis-position to acute anterior uveitis with ankylosing spondylitisrdquoInvestigative Ophthalmology amp Visual Science vol 55 no 12 pp7970ndash7974 2014
[20] T M Martin L Bye N Modi et al ldquoGenotype analysis ofpolymorphisms in autoimmune susceptibility genes CTLA-4and PTPN22 in an Acute anterior uveitis cohortrdquo MolecularVision vol 15 pp 208ndash212 2009
[21] Q Zhang J Qi S Hou et al ldquoA functional variant of PTPN22confers risk for Vogt-Koyanagi-Harada syndrome but not forankylosing spondylitisrdquo PLoS ONE vol 9 no 5 article e969432014
[22] S-S Zhao J-WHu JWang X-J Lou and L-L Zhou ldquoInversecorrelation between CD4+CD25 highCD127lowminus regulatory T-cells and serum immunoglobulin A in patients with new-onset ankylosing spondylitisrdquo Journal of International MedicalResearch vol 39 no 5 pp 1968ndash1974 2011
[23] W Zou Z Wu X Xiang S Sun and J Zhang ldquoThe expressionand significance of T helper cell subsets and regulatory Tcells CD4+CD25+ in peripheral blood of patients with humanleukocyte antigen B27-positive acute anterior uveitisrdquo GraefersquosArchive for Clinical and Experimental Ophthalmology vol 252no 4 pp 665ndash672 2014
BioMed Research International 9
[24] C Wang Q Liao Y Hu and D Zhong ldquoT lymphocyte subsetimbalances in patients contribute to ankylosing spondylitisrdquoExperimental and Therapeutic Medicine vol 9 no 1 pp 250ndash256 2015
[25] A Tedgui and Z Mallat ldquoCytokines in atherosclerosispathogenic and regulatory pathwaysrdquoPhysiological Reviews vol86 no 2 pp 515ndash581 2006
[26] P J Murray ldquoThe JAK-STAT signaling pathway Input andoutput integrationrdquo Journal of Immunology vol 178 no 5 pp2623ndash2629 2007
[27] A N Mathur H-C Chang D G Zisoulis et al ldquoStat3 andStat4 direct development of IL-17-secreting Th cellsrdquo Journal ofImmunology vol 178 no 8 pp 4901ndash4907 2007
[28] G K Hansson ldquoRegulation of immune mechanisms inatherosclerosisrdquo Annals of the New York Academy of Sciencesvol 947 pp 157ndash165 2001
[29] T Shea-Donohue A Fasano A Smith and A Zhao ldquoEntericpathogens and gut function role of cytokines and STATsrdquo GutMicrobes vol 1 no 5 pp 316ndash324 2010
[30] T Taniguchi K Ogasawara A Takaoka and N Tanaka ldquoIRFfamily of transcription factors as regulators of host defenserdquoAnnual Review of Immunology vol 19 pp 623ndash655 2001
[31] M Lohoff D Ferrick H-W Mittrucker et al ldquoInterferonregulatory factor-1 is required for a T helper 1 immune responsein vivordquo Immunity vol 6 no 6 pp 681ndash689 1997
[32] S Taki T Sato K Ogasawara et al ldquoMultistage regulation ofTh1-type immune responses by the transcription factor IRF-1rdquoImmunity vol 6 no 6 pp 673ndash679 1997
[33] C Bogdan ldquoNitric oxide and the immune responserdquo NatureImmunology vol 2 no 10 pp 907ndash916 2001
[34] S Brahmachari and K Pahan ldquoMyelin basic protein primingreduces the expression of Foxp3 in T cells via nitric oxiderdquoTheJournal of Immunology vol 184 no 4 pp 1799ndash1809 2010
[35] S-W Lee H Choi S-Y Eun S Fukuyama and M CroftldquoNitric oxide modulates TGF-120573-directive signals to suppressFoxp3 + regulatory T cell differentiation and potentiate Th1developmentrdquo Journal of Immunology vol 186 no 12 pp 6972ndash6980 2011
[36] D A Jabs R B Nussenblatt J T Rosenbaum and Standard-ization of Uveitis Nomenclature Working G ldquoStandardizationof uveitis nomenclature for reporting clinical data Resultsof the First International Workshoprdquo American Journal ofOphthalmology vol 140 pp 509ndash516 2005
[37] S Van Der Linden H A Valkenburg and A Cats ldquoEvaluationof diagnostic criteria for ankylosing spondylitis A proposal formodification of the New York criteriardquo Arthritis and Rheuma-tism vol 27 no 4 pp 361ndash368 1984
[38] D Ellinghaus E Ellinghaus R P Nair et al ldquoCombined anal-ysis of genome-wide association studies for Crohn disease andpsoriasis identifies seven shared susceptibility locirdquo AmericanJournal of Human Genetics vol 90 no 4 pp 636ndash647 2012
[39] G Fortunato G Calcagno V Bresciamorra et al ldquoMultiplesclerosis and hepatitis C virus infection are associated withsingle nucleotide polymorphisms in interferon pathway genesrdquoJournal of Interferon and Cytokine Research vol 28 no 3 pp141ndash152 2008
[40] W-H Hahn J-S Suh S H Cho B-S Cho and S-D KimldquoPolymorphisms of signal transducers and activators of tran-scription 1 and 4 (STAT1 and STAT4) contribute to progressionof childhood IgA nephropathyrdquo Cytokine vol 50 no 1 pp 69ndash74 2010
[41] S Hou J Qi Q Zhang et al ldquoGenetic variants in the JAK1 geneconfer higher risk of Behcetrsquos disease with ocular involvementin Han ChineserdquoHumanGenetics vol 132 no 9 pp 1049ndash10582013
[42] K Hu S Hou F Li Q Xiang A Kijlstra and P YangldquoJAK1 but not JAK2 and STAT3 confers susceptibility toVogt-Koyanagi-Harada (VKH) syndrome in a Han Chinesepopulationrdquo Investigative Ophthalmology and Visual Sciencevol 54 no 5 pp 3360ndash3365 2013
[43] A Cortes J Hadler J P Pointon et al ldquoIdentification ofmultiple risk variants for ankylosing spondylitis through high-density genotyping of immune-related locirdquo Nature Geneticsvol 45 no 7 pp 730ndash738 2013
[44] P A Shaiq P E Stuart A Latif et al ldquoGenetic associations ofpsoriasis in a Pakistani populationrdquo British Journal of Derma-tology vol 169 no 2 pp 406ndash411 2013
[45] S-K Yang Y Jung H Kim M Hong B D Ye and KSong ldquoAssociation of FCGR2A JAK2 or HNF4A variants withulcerative colitis in Koreansrdquo Digestive and Liver Disease vol43 no 11 pp 856ndash861 2011
[46] M Garcıa-Bermudez R Lopez-Mejıas F Genre et al ldquoLackof association between JAK3 gene polymorphisms and cardio-vascular disease in Spanish patients with rheumatoid arthritisrdquoBioMed Research International vol 2015 Article ID 318364 11pages 2015
[47] R Lopez-Mejıas M Garcıa-Bermudez C Gonzalez-Juanateyet al ldquoNFKB1-94ATTG insdel polymorphism (rs28362491) isassociated with cardiovascular disease in patients with rheuma-toid arthritisrdquo Atherosclerosis vol 224 no 2 pp 426ndash429 2012
[48] H J Park J W Kim B-S Cho and J-H Chung ldquoAssociationof BH3 interacting domain death agonist (BID) gene polymor-phisms with proteinuria of immunoglobulin A nephropathyrdquoScandinavian Journal of Clinical and Laboratory Investigationvol 74 no 4 pp 329ndash335 2014
Submit your manuscripts athttpwwwhindawicom
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Anatomy Research International
PeptidesInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporation httpwwwhindawicom
International Journal of
Volume 2014
Zoology
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Molecular Biology International
GenomicsInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
BioinformaticsAdvances in
Marine BiologyJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Signal TransductionJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
BioMed Research International
Evolutionary BiologyInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Biochemistry Research International
ArchaeaHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Genetics Research International
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Advances in
Virolog y
Hindawi Publishing Corporationhttpwwwhindawicom
Nucleic AcidsJournal of
Volume 2014
Stem CellsInternational
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Enzyme Research
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
International Journal of
Microbiology
BioMed Research International 7
differences in the genotype and allele frequencies of the other10 SNPs were observed between the HLA-B27-positive AS-positiveAAUpatients and healthy controls (119875Bonferroni gt 005Table 5) In addition there were no significant differencesin the genotype and allele frequencies of the tested SNPsbetween the AS-positive AAU patients and control subjects(119875Bonferroni gt 005 Supplementary Table S3)
33 Logistic Regression Analysis of SNPs in AAU We furtherinvestigated the SNPs rs10758669 and rs10975003 using addi-tive codominant dominant and recessive genetic modelsusing a multivariate logistic regression model adjusted forage and gender We observed that the frequency of the CAgenotype of rs10758669 was significantly higher in AAUpatients which suggests that patients with the rs10758669CA genotype have increased susceptibility to AAU (468versus 406 OR = 128 119875 = 002 SupplementaryTable S4) An increased frequency of the AC genotype ofrs10758669 was also observed in HLA-B27-positive AAUpatients and AS-positive AAU patients compared to thehealthy controls (495 versus 406 OR = 143 119875 =350 times 10minus3 and 492 versus 406 OR = 136 119875 = 002Supplementary Table S4) For SNP rs10975003 the frequencyof the heterozygous CT genotype was significantly higherin female AAU patients compared to the healthy controls(404 versus 336 OR = 157 119875 = 001 SupplementaryTable S5) A similar result was observed when we combinedCT and CC to construct a dominant model (443 versus380 OR = 157 119875 = 910times 10minus3 Supplementary Table S5)However none of the observed associations for the two SNPsretained statistical significance after Bonferroni correction(119875Bonferroni gt 005) Furthermore no significance associationswere found between the other SNPs and AAU even afterstratification by gender AS and HLA-B27 status (data notshown)
4 Discussion
In this study we first investigated whether genetic poly-morphisms of JAK-STAT signaling pathway genes includingJAK1 JAK2 STAT1 IRF1 and NOS2 confer susceptibilityto AAU with or without AS in a Chinese Han populationOur results suggest that none of these SNPs exhibit statisti-cally different frequencies of genotypes and alleles betweenhealthy controls and AAU patients However we observeda boundary significant association for two SNPs (rs10975003and rs10758669) of JAK2 by stratification analysis by genderand HLA-B27 status
We highlighted two issues at the time of study designto obtain unbiased association results First we followedstrict criteria for the diagnosis of AAU patients Patients withAAU were diagnosed as previously described by Jabs et al[36] and patients with AS were diagnosed with the modifiedNew York Criteria [37] In addition the AAU patients andhealthy controls were strictly matched by ethnicity and ageto avoid a possible influence of population stratificationFurthermore we only enrolled controls who had detailedhistories and physical examinations and excluded controlswith any autoimmune or immune-related diseases Finally
20 of the samples were randomly chosen and analyzed bydirect sequencing and the results of different genotypingmethods were consistent
AAU is defined as inflammation confined to the anteriorsegment of the eye that involves the iris and anterior part ofthe ciliary body The HLA-B27 is considered to be stronglyassociated with both AAU and AS [5 8 10 14] Recentgenetic studies have revealed that five candidate genes inthe JAK-STAT signaling pathway were considered geneticpredisposing factors for different autoimmune-mediated dis-eases [38ndash45] SNPs (rs10758669 and 10975003) in JAK2 areconsidered susceptibility factors for Crohnrsquos disease (CD) inthe German population and ulcerative colitis (UC) in theKorean population [38 45] One SNP (rs2070721) in IRF1 andSNPs (rs2297518 and rs4795067) in NOS2 are also associatedwith autoimmune diseases such as multiple sclerosis (MS)in Italy AS in Europe and psoriasis in Pakistan [39 43 44]Four SNPs (rs6718902 rs10199181 rs2066802 and rs1547550)in the STAT1 gene were observed to be associated with MSin Italy and IgA nephropathy (IgAN) in Korea [39 40] Inaddition an association was found between SNPs (rs2780815rs310241 rs3790532 rs310230 and rs310236) in JAK1 andBehcet disease (BD) as well as Vogt-Koyanagi-Harada (VKH)syndrome [41 42] two other common uveitis entities inChina There has been no report of associations betweenJAK-STAT signaling pathway genes and AAU and thus weperformed this case-control study to detect whether the fivecandidate genes were associated with AAU in a Chinese Hanpopulation Our results showed that there were no significantassociations between the genetic polymorphisms of the fivecandidate genes in the JAK-STAT signaling pathway andAAUThese results are not consistent with those observed forother autoimmune diseases reported in German Europeanand some other Asian populations [38 43 45] This discrep-ancy may be attributable to differences in the etiology andpathogenesis of AAU compared with BD VKH syndromeand autoimmune-mediated diseases
Consistent with our results a recent study also reportedno significant association of JAK-STAT signaling pathwaygene polymorphisms with rheumatoid arthritis stratified bythe presenceabsence of cardiovascular disease [46] Con-versely an influence of NFKB1 signaling pathway poly-morphisms on the development of cardiovascular events inpatients with rheumatoid arthritis has been observed [47]Furthermore NFKB1 signaling pathway polymorphismshave been described to play a critical role in the developmentof many autoimmune and inflammatory diseases and thusthe evaluation of the potential relationships between NFKB1polymorphisms and the development of AAU could be apromising research line for the future
There were several limitations of our studyWe had a lim-ited sample size to detect SNPs with weak effects while con-sidering multiple corrections Even for SNPs rs10758669 andrs10975003 we only had 700 power using a genetic powercalculator [48] In addition our samples were restricted to theHan Chinese population and all patients were enrolled fromthe ophthalmology department Further studies with a largersample size and other ethnic populations as well as patientsenrolled from multiple sources are warranted to confirm our
8 BioMed Research International
findings Additionally we only focused on eleven SNPs in theJAK-STAT pathway and it is possible that other unknownSNPs might be associated with AAU risk
In conclusion this study reveals that genetic polymor-phisms of JAK-STAT pathway genes including JAK1 JAK2STAT1 IRF1 andNOS2 may not be involved in susceptibilityto AAU risk in the Han Chinese population
Competing Interests
The authors declare that there is no conflict of interestsregarding the publication of this paper
Authorsrsquo Contributions
Ling Cheng and Hongsong Yu contributed equally to thiswork
Acknowledgments
The authors would like to thank Professor Peizeng Yang forhis great help in our study They are also grateful to alldonors enrolled in this study This work was supported bythe National Natural Science Foundation Project (8120067881670844) Fundamental and Advanced Research Programof Chongqing (cstc2015jcyjA10022) Science and TechnologyProject of Chongqing Municipal Education Commission(KJ1500236) Scientific Research Program of Science andTechnology Commission of Yuzhong District of Chongqing(20150102) and National Key Clinical Specialties Construc-tion Program of China
References
[1] D-K Hwang Y-J Chou C-Y Pu and P Chou ldquoEpidemi-ology of uveitis among the Chinese population in Taiwan APopulation-Based Studyrdquo Ophthalmology vol 119 no 11 pp2371ndash2376 2012
[2] K Nakao and N Ohba ldquoPrevalence of endogenous uveitis inKagoshima Prefecture Southwest JapanrdquoNipponGankaGakkaiZasshi vol 100 no 2 pp 150ndash155 1996
[3] D C Gritz and I GWong ldquoIncidence and prevalence of uveitisin Northern California the Northern California Epidemiologyof Uveitis Studyrdquo Ophthalmology vol 111 no 3 pp 491ndash5002004
[4] R B Nussenblatt and I Gery ldquoExperimental autoimmuneuveitis and its relationship to clinical ocular inflammatorydiseaserdquo Journal of Autoimmunity vol 9 no 5 pp 575ndash5851996
[5] P Yang Z Zhang H Zhou et al ldquoClinical patterns andcharacteristics of uveitis in a tertiary center for uveitis in ChinardquoCurrent Eye Research vol 30 no 11 pp 943ndash948 2005
[6] J T Gran and J F Skomsvoll ldquoThe outcome of ankylosingspondylitis a study of 100 patientsrdquo British Journal of Rheuma-tology vol 36 no 7 pp 766ndash771 1997
[7] L P Robertson andM J Davis ldquoA longitudinal study of diseaseactivity and functional status in a hospital cohort of patientswith ankylosing spondylitisrdquo Rheumatology vol 43 no 12 pp1565ndash1568 2004
[8] J H Chang P J McCluskey and D Wakefield ldquoAcute anterioruveitis and HLA-B27rdquo Survey of Ophthalmology vol 50 no 4pp 364ndash388 2005
[9] M Huhtinen and A Karma ldquoHLA-B27 typing in the categori-sation of uveitis in a HLA-B27 rich populationrdquo British Journalof Ophthalmology vol 84 no 4 pp 413ndash416 2000
[10] S Torres S Borges and A Artiles ldquoHLA-B27 and clinicalfeatures of acute anterior uveitis in Cubardquo Ocular Immunologyand Inflammation vol 21 no 2 pp 119ndash123 2013
[11] R Saari R Lahti K M Saari et al ldquoFrequency of rheumaticdiseases in patients with acute anterior uveitisrdquo ScandinavianJournal of Rheumatology vol 11 no 2 pp 121ndash123 1982
[12] A B Beckingsale J Davies J M Gibson and A RalphRosenthal ldquoAcute anterior uveitis ankylosing spondylitis backpain and HLA-B27rdquo British Journal of Ophthalmology vol 68no 10 pp 741ndash745 1984
[13] G P Thomas and M A Brown ldquoGenetics and genomics ofankylosing spondylitisrdquo Immunological Reviews vol 233 no 1pp 162ndash180 2010
[14] P C Robinson and M A Brown ldquoGenetics of ankylosingspondylitisrdquoMolecular Immunology vol 57 no 1 pp 2ndash11 2014
[15] C Lopez-Larrea K Sujirachato N K Mehra et al ldquoHLA-B27 subtypes in Asian patients with ankylosing spondylitisEvidence for new associationsrdquo Tissue Antigens vol 45 no 3pp 169ndash176 1995
[16] P C Robinson T A M Claushuis A Cortes et al ldquoGeneticdissection of acute anterior uveitis reveals similarities and dif-ferences in associations observed with ankylosing spondylitisrdquoArthritis and Rheumatology vol 67 no 1 pp 140ndash151 2015
[17] H Li S Hou H Yu et al ldquoAssociation of genetic variationsin TNFSF15 with acute anterior uveitis in Chinese HanrdquoInvestigative Ophthalmology and Visual Science vol 56 no 8pp 4605ndash4610 2015
[18] Q Xiang L Chen J Fang et al ldquoTNF receptor-associated factor5 gene confers genetic predisposition to acute anterior uveitisand pediatric uveitisrdquo Arthritis Research and Therapy vol 15no 5 article R113 2013
[19] H Yu Y Liu L Zhang et al ldquoFoxO1 gene confers genetic predis-position to acute anterior uveitis with ankylosing spondylitisrdquoInvestigative Ophthalmology amp Visual Science vol 55 no 12 pp7970ndash7974 2014
[20] T M Martin L Bye N Modi et al ldquoGenotype analysis ofpolymorphisms in autoimmune susceptibility genes CTLA-4and PTPN22 in an Acute anterior uveitis cohortrdquo MolecularVision vol 15 pp 208ndash212 2009
[21] Q Zhang J Qi S Hou et al ldquoA functional variant of PTPN22confers risk for Vogt-Koyanagi-Harada syndrome but not forankylosing spondylitisrdquo PLoS ONE vol 9 no 5 article e969432014
[22] S-S Zhao J-WHu JWang X-J Lou and L-L Zhou ldquoInversecorrelation between CD4+CD25 highCD127lowminus regulatory T-cells and serum immunoglobulin A in patients with new-onset ankylosing spondylitisrdquo Journal of International MedicalResearch vol 39 no 5 pp 1968ndash1974 2011
[23] W Zou Z Wu X Xiang S Sun and J Zhang ldquoThe expressionand significance of T helper cell subsets and regulatory Tcells CD4+CD25+ in peripheral blood of patients with humanleukocyte antigen B27-positive acute anterior uveitisrdquo GraefersquosArchive for Clinical and Experimental Ophthalmology vol 252no 4 pp 665ndash672 2014
BioMed Research International 9
[24] C Wang Q Liao Y Hu and D Zhong ldquoT lymphocyte subsetimbalances in patients contribute to ankylosing spondylitisrdquoExperimental and Therapeutic Medicine vol 9 no 1 pp 250ndash256 2015
[25] A Tedgui and Z Mallat ldquoCytokines in atherosclerosispathogenic and regulatory pathwaysrdquoPhysiological Reviews vol86 no 2 pp 515ndash581 2006
[26] P J Murray ldquoThe JAK-STAT signaling pathway Input andoutput integrationrdquo Journal of Immunology vol 178 no 5 pp2623ndash2629 2007
[27] A N Mathur H-C Chang D G Zisoulis et al ldquoStat3 andStat4 direct development of IL-17-secreting Th cellsrdquo Journal ofImmunology vol 178 no 8 pp 4901ndash4907 2007
[28] G K Hansson ldquoRegulation of immune mechanisms inatherosclerosisrdquo Annals of the New York Academy of Sciencesvol 947 pp 157ndash165 2001
[29] T Shea-Donohue A Fasano A Smith and A Zhao ldquoEntericpathogens and gut function role of cytokines and STATsrdquo GutMicrobes vol 1 no 5 pp 316ndash324 2010
[30] T Taniguchi K Ogasawara A Takaoka and N Tanaka ldquoIRFfamily of transcription factors as regulators of host defenserdquoAnnual Review of Immunology vol 19 pp 623ndash655 2001
[31] M Lohoff D Ferrick H-W Mittrucker et al ldquoInterferonregulatory factor-1 is required for a T helper 1 immune responsein vivordquo Immunity vol 6 no 6 pp 681ndash689 1997
[32] S Taki T Sato K Ogasawara et al ldquoMultistage regulation ofTh1-type immune responses by the transcription factor IRF-1rdquoImmunity vol 6 no 6 pp 673ndash679 1997
[33] C Bogdan ldquoNitric oxide and the immune responserdquo NatureImmunology vol 2 no 10 pp 907ndash916 2001
[34] S Brahmachari and K Pahan ldquoMyelin basic protein primingreduces the expression of Foxp3 in T cells via nitric oxiderdquoTheJournal of Immunology vol 184 no 4 pp 1799ndash1809 2010
[35] S-W Lee H Choi S-Y Eun S Fukuyama and M CroftldquoNitric oxide modulates TGF-120573-directive signals to suppressFoxp3 + regulatory T cell differentiation and potentiate Th1developmentrdquo Journal of Immunology vol 186 no 12 pp 6972ndash6980 2011
[36] D A Jabs R B Nussenblatt J T Rosenbaum and Standard-ization of Uveitis Nomenclature Working G ldquoStandardizationof uveitis nomenclature for reporting clinical data Resultsof the First International Workshoprdquo American Journal ofOphthalmology vol 140 pp 509ndash516 2005
[37] S Van Der Linden H A Valkenburg and A Cats ldquoEvaluationof diagnostic criteria for ankylosing spondylitis A proposal formodification of the New York criteriardquo Arthritis and Rheuma-tism vol 27 no 4 pp 361ndash368 1984
[38] D Ellinghaus E Ellinghaus R P Nair et al ldquoCombined anal-ysis of genome-wide association studies for Crohn disease andpsoriasis identifies seven shared susceptibility locirdquo AmericanJournal of Human Genetics vol 90 no 4 pp 636ndash647 2012
[39] G Fortunato G Calcagno V Bresciamorra et al ldquoMultiplesclerosis and hepatitis C virus infection are associated withsingle nucleotide polymorphisms in interferon pathway genesrdquoJournal of Interferon and Cytokine Research vol 28 no 3 pp141ndash152 2008
[40] W-H Hahn J-S Suh S H Cho B-S Cho and S-D KimldquoPolymorphisms of signal transducers and activators of tran-scription 1 and 4 (STAT1 and STAT4) contribute to progressionof childhood IgA nephropathyrdquo Cytokine vol 50 no 1 pp 69ndash74 2010
[41] S Hou J Qi Q Zhang et al ldquoGenetic variants in the JAK1 geneconfer higher risk of Behcetrsquos disease with ocular involvementin Han ChineserdquoHumanGenetics vol 132 no 9 pp 1049ndash10582013
[42] K Hu S Hou F Li Q Xiang A Kijlstra and P YangldquoJAK1 but not JAK2 and STAT3 confers susceptibility toVogt-Koyanagi-Harada (VKH) syndrome in a Han Chinesepopulationrdquo Investigative Ophthalmology and Visual Sciencevol 54 no 5 pp 3360ndash3365 2013
[43] A Cortes J Hadler J P Pointon et al ldquoIdentification ofmultiple risk variants for ankylosing spondylitis through high-density genotyping of immune-related locirdquo Nature Geneticsvol 45 no 7 pp 730ndash738 2013
[44] P A Shaiq P E Stuart A Latif et al ldquoGenetic associations ofpsoriasis in a Pakistani populationrdquo British Journal of Derma-tology vol 169 no 2 pp 406ndash411 2013
[45] S-K Yang Y Jung H Kim M Hong B D Ye and KSong ldquoAssociation of FCGR2A JAK2 or HNF4A variants withulcerative colitis in Koreansrdquo Digestive and Liver Disease vol43 no 11 pp 856ndash861 2011
[46] M Garcıa-Bermudez R Lopez-Mejıas F Genre et al ldquoLackof association between JAK3 gene polymorphisms and cardio-vascular disease in Spanish patients with rheumatoid arthritisrdquoBioMed Research International vol 2015 Article ID 318364 11pages 2015
[47] R Lopez-Mejıas M Garcıa-Bermudez C Gonzalez-Juanateyet al ldquoNFKB1-94ATTG insdel polymorphism (rs28362491) isassociated with cardiovascular disease in patients with rheuma-toid arthritisrdquo Atherosclerosis vol 224 no 2 pp 426ndash429 2012
[48] H J Park J W Kim B-S Cho and J-H Chung ldquoAssociationof BH3 interacting domain death agonist (BID) gene polymor-phisms with proteinuria of immunoglobulin A nephropathyrdquoScandinavian Journal of Clinical and Laboratory Investigationvol 74 no 4 pp 329ndash335 2014
Submit your manuscripts athttpwwwhindawicom
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Anatomy Research International
PeptidesInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporation httpwwwhindawicom
International Journal of
Volume 2014
Zoology
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Molecular Biology International
GenomicsInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
BioinformaticsAdvances in
Marine BiologyJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Signal TransductionJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
BioMed Research International
Evolutionary BiologyInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Biochemistry Research International
ArchaeaHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Genetics Research International
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Advances in
Virolog y
Hindawi Publishing Corporationhttpwwwhindawicom
Nucleic AcidsJournal of
Volume 2014
Stem CellsInternational
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Enzyme Research
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
International Journal of
Microbiology
8 BioMed Research International
findings Additionally we only focused on eleven SNPs in theJAK-STAT pathway and it is possible that other unknownSNPs might be associated with AAU risk
In conclusion this study reveals that genetic polymor-phisms of JAK-STAT pathway genes including JAK1 JAK2STAT1 IRF1 andNOS2 may not be involved in susceptibilityto AAU risk in the Han Chinese population
Competing Interests
The authors declare that there is no conflict of interestsregarding the publication of this paper
Authorsrsquo Contributions
Ling Cheng and Hongsong Yu contributed equally to thiswork
Acknowledgments
The authors would like to thank Professor Peizeng Yang forhis great help in our study They are also grateful to alldonors enrolled in this study This work was supported bythe National Natural Science Foundation Project (8120067881670844) Fundamental and Advanced Research Programof Chongqing (cstc2015jcyjA10022) Science and TechnologyProject of Chongqing Municipal Education Commission(KJ1500236) Scientific Research Program of Science andTechnology Commission of Yuzhong District of Chongqing(20150102) and National Key Clinical Specialties Construc-tion Program of China
References
[1] D-K Hwang Y-J Chou C-Y Pu and P Chou ldquoEpidemi-ology of uveitis among the Chinese population in Taiwan APopulation-Based Studyrdquo Ophthalmology vol 119 no 11 pp2371ndash2376 2012
[2] K Nakao and N Ohba ldquoPrevalence of endogenous uveitis inKagoshima Prefecture Southwest JapanrdquoNipponGankaGakkaiZasshi vol 100 no 2 pp 150ndash155 1996
[3] D C Gritz and I GWong ldquoIncidence and prevalence of uveitisin Northern California the Northern California Epidemiologyof Uveitis Studyrdquo Ophthalmology vol 111 no 3 pp 491ndash5002004
[4] R B Nussenblatt and I Gery ldquoExperimental autoimmuneuveitis and its relationship to clinical ocular inflammatorydiseaserdquo Journal of Autoimmunity vol 9 no 5 pp 575ndash5851996
[5] P Yang Z Zhang H Zhou et al ldquoClinical patterns andcharacteristics of uveitis in a tertiary center for uveitis in ChinardquoCurrent Eye Research vol 30 no 11 pp 943ndash948 2005
[6] J T Gran and J F Skomsvoll ldquoThe outcome of ankylosingspondylitis a study of 100 patientsrdquo British Journal of Rheuma-tology vol 36 no 7 pp 766ndash771 1997
[7] L P Robertson andM J Davis ldquoA longitudinal study of diseaseactivity and functional status in a hospital cohort of patientswith ankylosing spondylitisrdquo Rheumatology vol 43 no 12 pp1565ndash1568 2004
[8] J H Chang P J McCluskey and D Wakefield ldquoAcute anterioruveitis and HLA-B27rdquo Survey of Ophthalmology vol 50 no 4pp 364ndash388 2005
[9] M Huhtinen and A Karma ldquoHLA-B27 typing in the categori-sation of uveitis in a HLA-B27 rich populationrdquo British Journalof Ophthalmology vol 84 no 4 pp 413ndash416 2000
[10] S Torres S Borges and A Artiles ldquoHLA-B27 and clinicalfeatures of acute anterior uveitis in Cubardquo Ocular Immunologyand Inflammation vol 21 no 2 pp 119ndash123 2013
[11] R Saari R Lahti K M Saari et al ldquoFrequency of rheumaticdiseases in patients with acute anterior uveitisrdquo ScandinavianJournal of Rheumatology vol 11 no 2 pp 121ndash123 1982
[12] A B Beckingsale J Davies J M Gibson and A RalphRosenthal ldquoAcute anterior uveitis ankylosing spondylitis backpain and HLA-B27rdquo British Journal of Ophthalmology vol 68no 10 pp 741ndash745 1984
[13] G P Thomas and M A Brown ldquoGenetics and genomics ofankylosing spondylitisrdquo Immunological Reviews vol 233 no 1pp 162ndash180 2010
[14] P C Robinson and M A Brown ldquoGenetics of ankylosingspondylitisrdquoMolecular Immunology vol 57 no 1 pp 2ndash11 2014
[15] C Lopez-Larrea K Sujirachato N K Mehra et al ldquoHLA-B27 subtypes in Asian patients with ankylosing spondylitisEvidence for new associationsrdquo Tissue Antigens vol 45 no 3pp 169ndash176 1995
[16] P C Robinson T A M Claushuis A Cortes et al ldquoGeneticdissection of acute anterior uveitis reveals similarities and dif-ferences in associations observed with ankylosing spondylitisrdquoArthritis and Rheumatology vol 67 no 1 pp 140ndash151 2015
[17] H Li S Hou H Yu et al ldquoAssociation of genetic variationsin TNFSF15 with acute anterior uveitis in Chinese HanrdquoInvestigative Ophthalmology and Visual Science vol 56 no 8pp 4605ndash4610 2015
[18] Q Xiang L Chen J Fang et al ldquoTNF receptor-associated factor5 gene confers genetic predisposition to acute anterior uveitisand pediatric uveitisrdquo Arthritis Research and Therapy vol 15no 5 article R113 2013
[19] H Yu Y Liu L Zhang et al ldquoFoxO1 gene confers genetic predis-position to acute anterior uveitis with ankylosing spondylitisrdquoInvestigative Ophthalmology amp Visual Science vol 55 no 12 pp7970ndash7974 2014
[20] T M Martin L Bye N Modi et al ldquoGenotype analysis ofpolymorphisms in autoimmune susceptibility genes CTLA-4and PTPN22 in an Acute anterior uveitis cohortrdquo MolecularVision vol 15 pp 208ndash212 2009
[21] Q Zhang J Qi S Hou et al ldquoA functional variant of PTPN22confers risk for Vogt-Koyanagi-Harada syndrome but not forankylosing spondylitisrdquo PLoS ONE vol 9 no 5 article e969432014
[22] S-S Zhao J-WHu JWang X-J Lou and L-L Zhou ldquoInversecorrelation between CD4+CD25 highCD127lowminus regulatory T-cells and serum immunoglobulin A in patients with new-onset ankylosing spondylitisrdquo Journal of International MedicalResearch vol 39 no 5 pp 1968ndash1974 2011
[23] W Zou Z Wu X Xiang S Sun and J Zhang ldquoThe expressionand significance of T helper cell subsets and regulatory Tcells CD4+CD25+ in peripheral blood of patients with humanleukocyte antigen B27-positive acute anterior uveitisrdquo GraefersquosArchive for Clinical and Experimental Ophthalmology vol 252no 4 pp 665ndash672 2014
BioMed Research International 9
[24] C Wang Q Liao Y Hu and D Zhong ldquoT lymphocyte subsetimbalances in patients contribute to ankylosing spondylitisrdquoExperimental and Therapeutic Medicine vol 9 no 1 pp 250ndash256 2015
[25] A Tedgui and Z Mallat ldquoCytokines in atherosclerosispathogenic and regulatory pathwaysrdquoPhysiological Reviews vol86 no 2 pp 515ndash581 2006
[26] P J Murray ldquoThe JAK-STAT signaling pathway Input andoutput integrationrdquo Journal of Immunology vol 178 no 5 pp2623ndash2629 2007
[27] A N Mathur H-C Chang D G Zisoulis et al ldquoStat3 andStat4 direct development of IL-17-secreting Th cellsrdquo Journal ofImmunology vol 178 no 8 pp 4901ndash4907 2007
[28] G K Hansson ldquoRegulation of immune mechanisms inatherosclerosisrdquo Annals of the New York Academy of Sciencesvol 947 pp 157ndash165 2001
[29] T Shea-Donohue A Fasano A Smith and A Zhao ldquoEntericpathogens and gut function role of cytokines and STATsrdquo GutMicrobes vol 1 no 5 pp 316ndash324 2010
[30] T Taniguchi K Ogasawara A Takaoka and N Tanaka ldquoIRFfamily of transcription factors as regulators of host defenserdquoAnnual Review of Immunology vol 19 pp 623ndash655 2001
[31] M Lohoff D Ferrick H-W Mittrucker et al ldquoInterferonregulatory factor-1 is required for a T helper 1 immune responsein vivordquo Immunity vol 6 no 6 pp 681ndash689 1997
[32] S Taki T Sato K Ogasawara et al ldquoMultistage regulation ofTh1-type immune responses by the transcription factor IRF-1rdquoImmunity vol 6 no 6 pp 673ndash679 1997
[33] C Bogdan ldquoNitric oxide and the immune responserdquo NatureImmunology vol 2 no 10 pp 907ndash916 2001
[34] S Brahmachari and K Pahan ldquoMyelin basic protein primingreduces the expression of Foxp3 in T cells via nitric oxiderdquoTheJournal of Immunology vol 184 no 4 pp 1799ndash1809 2010
[35] S-W Lee H Choi S-Y Eun S Fukuyama and M CroftldquoNitric oxide modulates TGF-120573-directive signals to suppressFoxp3 + regulatory T cell differentiation and potentiate Th1developmentrdquo Journal of Immunology vol 186 no 12 pp 6972ndash6980 2011
[36] D A Jabs R B Nussenblatt J T Rosenbaum and Standard-ization of Uveitis Nomenclature Working G ldquoStandardizationof uveitis nomenclature for reporting clinical data Resultsof the First International Workshoprdquo American Journal ofOphthalmology vol 140 pp 509ndash516 2005
[37] S Van Der Linden H A Valkenburg and A Cats ldquoEvaluationof diagnostic criteria for ankylosing spondylitis A proposal formodification of the New York criteriardquo Arthritis and Rheuma-tism vol 27 no 4 pp 361ndash368 1984
[38] D Ellinghaus E Ellinghaus R P Nair et al ldquoCombined anal-ysis of genome-wide association studies for Crohn disease andpsoriasis identifies seven shared susceptibility locirdquo AmericanJournal of Human Genetics vol 90 no 4 pp 636ndash647 2012
[39] G Fortunato G Calcagno V Bresciamorra et al ldquoMultiplesclerosis and hepatitis C virus infection are associated withsingle nucleotide polymorphisms in interferon pathway genesrdquoJournal of Interferon and Cytokine Research vol 28 no 3 pp141ndash152 2008
[40] W-H Hahn J-S Suh S H Cho B-S Cho and S-D KimldquoPolymorphisms of signal transducers and activators of tran-scription 1 and 4 (STAT1 and STAT4) contribute to progressionof childhood IgA nephropathyrdquo Cytokine vol 50 no 1 pp 69ndash74 2010
[41] S Hou J Qi Q Zhang et al ldquoGenetic variants in the JAK1 geneconfer higher risk of Behcetrsquos disease with ocular involvementin Han ChineserdquoHumanGenetics vol 132 no 9 pp 1049ndash10582013
[42] K Hu S Hou F Li Q Xiang A Kijlstra and P YangldquoJAK1 but not JAK2 and STAT3 confers susceptibility toVogt-Koyanagi-Harada (VKH) syndrome in a Han Chinesepopulationrdquo Investigative Ophthalmology and Visual Sciencevol 54 no 5 pp 3360ndash3365 2013
[43] A Cortes J Hadler J P Pointon et al ldquoIdentification ofmultiple risk variants for ankylosing spondylitis through high-density genotyping of immune-related locirdquo Nature Geneticsvol 45 no 7 pp 730ndash738 2013
[44] P A Shaiq P E Stuart A Latif et al ldquoGenetic associations ofpsoriasis in a Pakistani populationrdquo British Journal of Derma-tology vol 169 no 2 pp 406ndash411 2013
[45] S-K Yang Y Jung H Kim M Hong B D Ye and KSong ldquoAssociation of FCGR2A JAK2 or HNF4A variants withulcerative colitis in Koreansrdquo Digestive and Liver Disease vol43 no 11 pp 856ndash861 2011
[46] M Garcıa-Bermudez R Lopez-Mejıas F Genre et al ldquoLackof association between JAK3 gene polymorphisms and cardio-vascular disease in Spanish patients with rheumatoid arthritisrdquoBioMed Research International vol 2015 Article ID 318364 11pages 2015
[47] R Lopez-Mejıas M Garcıa-Bermudez C Gonzalez-Juanateyet al ldquoNFKB1-94ATTG insdel polymorphism (rs28362491) isassociated with cardiovascular disease in patients with rheuma-toid arthritisrdquo Atherosclerosis vol 224 no 2 pp 426ndash429 2012
[48] H J Park J W Kim B-S Cho and J-H Chung ldquoAssociationof BH3 interacting domain death agonist (BID) gene polymor-phisms with proteinuria of immunoglobulin A nephropathyrdquoScandinavian Journal of Clinical and Laboratory Investigationvol 74 no 4 pp 329ndash335 2014
Submit your manuscripts athttpwwwhindawicom
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Anatomy Research International
PeptidesInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporation httpwwwhindawicom
International Journal of
Volume 2014
Zoology
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Molecular Biology International
GenomicsInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
BioinformaticsAdvances in
Marine BiologyJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Signal TransductionJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
BioMed Research International
Evolutionary BiologyInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Biochemistry Research International
ArchaeaHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Genetics Research International
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Advances in
Virolog y
Hindawi Publishing Corporationhttpwwwhindawicom
Nucleic AcidsJournal of
Volume 2014
Stem CellsInternational
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Enzyme Research
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
International Journal of
Microbiology
BioMed Research International 9
[24] C Wang Q Liao Y Hu and D Zhong ldquoT lymphocyte subsetimbalances in patients contribute to ankylosing spondylitisrdquoExperimental and Therapeutic Medicine vol 9 no 1 pp 250ndash256 2015
[25] A Tedgui and Z Mallat ldquoCytokines in atherosclerosispathogenic and regulatory pathwaysrdquoPhysiological Reviews vol86 no 2 pp 515ndash581 2006
[26] P J Murray ldquoThe JAK-STAT signaling pathway Input andoutput integrationrdquo Journal of Immunology vol 178 no 5 pp2623ndash2629 2007
[27] A N Mathur H-C Chang D G Zisoulis et al ldquoStat3 andStat4 direct development of IL-17-secreting Th cellsrdquo Journal ofImmunology vol 178 no 8 pp 4901ndash4907 2007
[28] G K Hansson ldquoRegulation of immune mechanisms inatherosclerosisrdquo Annals of the New York Academy of Sciencesvol 947 pp 157ndash165 2001
[29] T Shea-Donohue A Fasano A Smith and A Zhao ldquoEntericpathogens and gut function role of cytokines and STATsrdquo GutMicrobes vol 1 no 5 pp 316ndash324 2010
[30] T Taniguchi K Ogasawara A Takaoka and N Tanaka ldquoIRFfamily of transcription factors as regulators of host defenserdquoAnnual Review of Immunology vol 19 pp 623ndash655 2001
[31] M Lohoff D Ferrick H-W Mittrucker et al ldquoInterferonregulatory factor-1 is required for a T helper 1 immune responsein vivordquo Immunity vol 6 no 6 pp 681ndash689 1997
[32] S Taki T Sato K Ogasawara et al ldquoMultistage regulation ofTh1-type immune responses by the transcription factor IRF-1rdquoImmunity vol 6 no 6 pp 673ndash679 1997
[33] C Bogdan ldquoNitric oxide and the immune responserdquo NatureImmunology vol 2 no 10 pp 907ndash916 2001
[34] S Brahmachari and K Pahan ldquoMyelin basic protein primingreduces the expression of Foxp3 in T cells via nitric oxiderdquoTheJournal of Immunology vol 184 no 4 pp 1799ndash1809 2010
[35] S-W Lee H Choi S-Y Eun S Fukuyama and M CroftldquoNitric oxide modulates TGF-120573-directive signals to suppressFoxp3 + regulatory T cell differentiation and potentiate Th1developmentrdquo Journal of Immunology vol 186 no 12 pp 6972ndash6980 2011
[36] D A Jabs R B Nussenblatt J T Rosenbaum and Standard-ization of Uveitis Nomenclature Working G ldquoStandardizationof uveitis nomenclature for reporting clinical data Resultsof the First International Workshoprdquo American Journal ofOphthalmology vol 140 pp 509ndash516 2005
[37] S Van Der Linden H A Valkenburg and A Cats ldquoEvaluationof diagnostic criteria for ankylosing spondylitis A proposal formodification of the New York criteriardquo Arthritis and Rheuma-tism vol 27 no 4 pp 361ndash368 1984
[38] D Ellinghaus E Ellinghaus R P Nair et al ldquoCombined anal-ysis of genome-wide association studies for Crohn disease andpsoriasis identifies seven shared susceptibility locirdquo AmericanJournal of Human Genetics vol 90 no 4 pp 636ndash647 2012
[39] G Fortunato G Calcagno V Bresciamorra et al ldquoMultiplesclerosis and hepatitis C virus infection are associated withsingle nucleotide polymorphisms in interferon pathway genesrdquoJournal of Interferon and Cytokine Research vol 28 no 3 pp141ndash152 2008
[40] W-H Hahn J-S Suh S H Cho B-S Cho and S-D KimldquoPolymorphisms of signal transducers and activators of tran-scription 1 and 4 (STAT1 and STAT4) contribute to progressionof childhood IgA nephropathyrdquo Cytokine vol 50 no 1 pp 69ndash74 2010
[41] S Hou J Qi Q Zhang et al ldquoGenetic variants in the JAK1 geneconfer higher risk of Behcetrsquos disease with ocular involvementin Han ChineserdquoHumanGenetics vol 132 no 9 pp 1049ndash10582013
[42] K Hu S Hou F Li Q Xiang A Kijlstra and P YangldquoJAK1 but not JAK2 and STAT3 confers susceptibility toVogt-Koyanagi-Harada (VKH) syndrome in a Han Chinesepopulationrdquo Investigative Ophthalmology and Visual Sciencevol 54 no 5 pp 3360ndash3365 2013
[43] A Cortes J Hadler J P Pointon et al ldquoIdentification ofmultiple risk variants for ankylosing spondylitis through high-density genotyping of immune-related locirdquo Nature Geneticsvol 45 no 7 pp 730ndash738 2013
[44] P A Shaiq P E Stuart A Latif et al ldquoGenetic associations ofpsoriasis in a Pakistani populationrdquo British Journal of Derma-tology vol 169 no 2 pp 406ndash411 2013
[45] S-K Yang Y Jung H Kim M Hong B D Ye and KSong ldquoAssociation of FCGR2A JAK2 or HNF4A variants withulcerative colitis in Koreansrdquo Digestive and Liver Disease vol43 no 11 pp 856ndash861 2011
[46] M Garcıa-Bermudez R Lopez-Mejıas F Genre et al ldquoLackof association between JAK3 gene polymorphisms and cardio-vascular disease in Spanish patients with rheumatoid arthritisrdquoBioMed Research International vol 2015 Article ID 318364 11pages 2015
[47] R Lopez-Mejıas M Garcıa-Bermudez C Gonzalez-Juanateyet al ldquoNFKB1-94ATTG insdel polymorphism (rs28362491) isassociated with cardiovascular disease in patients with rheuma-toid arthritisrdquo Atherosclerosis vol 224 no 2 pp 426ndash429 2012
[48] H J Park J W Kim B-S Cho and J-H Chung ldquoAssociationof BH3 interacting domain death agonist (BID) gene polymor-phisms with proteinuria of immunoglobulin A nephropathyrdquoScandinavian Journal of Clinical and Laboratory Investigationvol 74 no 4 pp 329ndash335 2014
Submit your manuscripts athttpwwwhindawicom
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Anatomy Research International
PeptidesInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporation httpwwwhindawicom
International Journal of
Volume 2014
Zoology
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Molecular Biology International
GenomicsInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
BioinformaticsAdvances in
Marine BiologyJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Signal TransductionJournal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
BioMed Research International
Evolutionary BiologyInternational Journal of
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Biochemistry Research International
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ArchaeaHindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Genetics Research International
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Advances in
Virolog y
Hindawi Publishing Corporationhttpwwwhindawicom
Nucleic AcidsJournal of
Volume 2014
Stem CellsInternational
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
Enzyme Research
Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014
International Journal of
Microbiology
