Salivary Cytokines Coincide with Disease Characteristic in …… · 2021. 4. 18. · Emily A....
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EE Starman, PM Gomez Hernandez, AB Davis, F Qian, SM Lieberman, KA Brogden, EA Lanzel University of Iowa, Iowa City, IA, USA Salivary Cytokines Coincide with Disease Characteristic in Childhood Sjögren Syndrome Introduction Sjögren syndrome is an autoimmune disease that targets that salivary and lacrimal glands (Figure 1). Though most prevalent in females in their 4th-5th decade, the disease can also occur in chil- dren. Assessing the chemokine, cytokine, and biomarker (CCBM) profiles in these individuals may help identify unknown early events in the etiopathogenesis of both childhood/adult Sjögren syn- drome. We hypothesized that unique profiles of CCBMs in saliva associated with CD4 + T cell, CD8 + T cell, B cell, and mononuclear cell infiltrates may coincide with signs and symptoms of the disease process. Signs and symptoms of disease include: • Focal sialadenitis • Presence of auto-antibodies • SS-A • SS-B • Rheumatoid Factor • Dry eye symptoms • Dry mouth symptoms • Recurrent acute parotitis Materials and Methods Conclusions • The results in this study suggest that B2M, MMP9, and S100A8 profiled in saliva of children with Sjӧgren Syndrome (SS) may coincide with the characteristics of their disease. • There were no benefits observed due to diluting saliva samples prior to analysis. • These findings may have an important impact on establishing more objective and efficient diagnosis of Sjögren Syndrome, allowing for earlier therapeutic intervention and improved disease management. Possible therapeutic targets may also be identified through analyzing saliva profiles. Results • Twenty-seven subjects (88.9% females) were included in the study, and the mean age was 15.4±3.1 years with a range of 10-20 years. • Signs and symptoms of children diagnosed with SS were compared to children with SS without signs and symptoms and to healthy, control patients • There was a statistically significant difference in concentrations of B2M (P=.002; the Wilcoxon rank-sum test), MMP9 (P=.001; the Wilcoxon rank-sum test), and S100A8 (P=.005; the Wilcoxon rank-sum test) between subjects with dry eye symptoms (n=8) and those without dry eye symp - toms (n=19) (Figure 2). • There was a statistically significant difference in concentrations of MMP9 (P=.038; the Wilcox - on rank-sum test) between subjects with dry mouth symptoms (n=6) and those without dry mouth symptoms (n=21) (Figure 3). • There was a statistically significant difference in concentrations of MMP9 (P=.042; a one-way ANOVA) among 3 groups of subjects with different levels of focal sialadenitis: none (n=18), +, foci present, but focus score not reported or <1 focus/4mm2 (n=2), and ++, focus score ≥ 1 focus/4mm 2 (n= 7) (Figure 4). • There were no differences in concentrations of 7 markers between children with and without SS-A antibody, SS-B antibody, RF-antibody and acute parotitis. Figure 1. H and E stain of a salivary gland biopsy from SS patient courtesy of Dr. Lanzel CCBM Description and function A2M (alpha 2-Macroglobulin) A protease inhibitor that also inhibits inflammatory cytokines and disrupts inflammatory cascades AMBP (alpha 1-Microglobulin) Transport proteins and plays a role in the regulation of inflammatory processes B2M (beta 2-Microglobulin) Transmembrance receptor found in association with the histocompatibility complex C9 (complement component 9) Final component of the complement system MMP9 (matrix metallopeptidase 9) A peptidase with pro-inflammatory/anti- inflammatory activities S100A8 (S100 calcium binding protein A8) Involved in the regulation of a number of cellular processes such as cell cycle progression and differentiation SLPI (Secretory leukocyte protease inhibitor) A secreted inhibitor, which protects epithelial tissues from attack by endogenous proteolytic enzymes Table 1. Selected CCBMs and their functions Acknowledgements Supported by the Pilot Research Award from the Sjӧgren Syndrome Foundation and the University of Iowa College of Dentistry Dental Student Research Program College of Dentistry, University of Iowa; Paula M. Gomez Hernandez, Fang Qian, Erliang Zeng, Kim A. Brogden, Emily A. Lanzel Carver College of Medicine, University of Iowa; Scott M. Lieberman and Andrew B. Davis University of Iowa Tissue Procurement Core Statistical Analysis • One way ANOVA with post‐ hoc Tukey‐Kramer test •Nonpapmetric Wilcoxon rank‐sum test •Kruskal‐Wallis text with post‐hoc Bonferroni adjustment test •P‐value <0.05 was used as criteria for statistical significance and 0.05<p<0.20 was used for marginal significance Determine CCBM concentrations • 7 CCBMs selected (A2M, AMBP, B2M, C9, MMP9, S100A8, and SLPI) based on their roles in immune reactions and high concentrations in previous study • Multiplex fluorescent bead‐based immunoassays: Luminex xMAP Collect and prepare saliva samples • 11 children diagnosed with and managing Sjögren Syndrome • 16 gender and age matched healthy controls Figure 2. B2M, MMP9, and S100A8 concentrations between subjects with and without dry eye symptoms. Figure 3. MMP9 concentrations between subjects with and without dry mouth symptoms Figure 4. MMP9 concentrations among 3 groups of subjects with different levels of focal sialadenitis 0 100000 200000 300000 400000 500000 600000 700000 800000 900000 1000000 B2M MMP9 S100A8 Concentration (pg/ml) Dry eye No Dry eye 1 10 100 1000 10000 100000 MMP9 Log(concentration (pg/ml)) Dry Mouth No Dry Mouth 0 50000 100000 150000 200000 250000 300000 350000 400000 450000 500000 MMP9 Concentration (pg/ml) No Focal Sialadenitis (+) (++)
Transcript of Salivary Cytokines Coincide with Disease Characteristic in …… · 2021. 4. 18. · Emily A....
EE Starman, PM Gomez Hernandez, AB Davis, F Qian, SM Lieberman, KA Brogden, EA LanzelUniversity of Iowa, Iowa City, IA, USA
Salivary Cytokines Coincide with Disease Characteristic in Childhood Sjögren Syndrome
IntroductionSjögren syndrome is an autoimmune disease that targets that salivary and lacrimal glands (Figure 1). Though most prevalent in females in their 4th-5th decade, the disease can also occur in chil-dren. Assessing the chemokine, cytokine, and biomarker (CCBM) profiles in these individuals may help identify unknown early events in the etiopathogenesis of both childhood/adult Sjögren syn-drome. We hypothesized that unique profiles of CCBMs in saliva associated with CD4+ T cell, CD8+ T cell, B cell, and mononuclear cell infiltrates may coincide with signs and symptoms of the disease process.
Signs and symptoms of disease include:• Focal sialadenitis• Presence of auto-antibodies
• SS-A• SS-B• Rheumatoid Factor
• Dry eye symptoms• Dry mouth symptoms• Recurrent acute parotitis
Materials and Methods Conclusions• The results in this study suggest that B2M, MMP9, and S100A8 profiled in saliva of children
with Sjӧgren Syndrome (SS) may coincide with the characteristics of their disease.• There were no benefits observed due to diluting saliva samples prior to analysis.• These findings may have an important impact on establishing more objective and efficient
diagnosis of Sjögren Syndrome, allowing for earlier therapeutic intervention and improved disease management. Possible therapeutic targets may also be identified through analyzing saliva profiles.
Results• Twenty-seven subjects (88.9% females) were included in the study, and the mean age was
15.4±3.1 years with a range of 10-20 years.• Signs and symptoms of children diagnosed with SS were compared to children with SS without
signs and symptoms and to healthy, control patients • There was a statistically significant difference in concentrations of B2M (P=.002; the Wilcoxon
rank-sum test), MMP9 (P=.001; the Wilcoxon rank-sum test), and S100A8 (P=.005; the Wilcoxon rank-sum test) between subjects with dry eye symptoms (n=8) and those without dry eye symp-toms (n=19) (Figure 2).
• There was a statistically significant difference in concentrations of MMP9 (P=.038; the Wilcox-on rank-sum test) between subjects with dry mouth symptoms (n=6) and those without dry mouth symptoms (n=21) (Figure 3).
• There was a statistically significant difference in concentrations of MMP9 (P=.042; a one-way ANOVA) among 3 groups of subjects with different levels of focal sialadenitis: none (n=18), +, foci present, but focus score not reported or <1 focus/4mm2 (n=2), and ++, focus score ≥ 1 focus/4mm2 (n= 7) (Figure 4).
• There were no differences in concentrations of 7 markers between children with and without SS-A antibody, SS-B antibody, RF-antibody and acute parotitis.
Figure 1. H and E stain of a salivary gland biopsy from SS patient courtesy of Dr. Lanzel
CCBM Description and function
A2M (alpha 2-Macroglobulin) A protease inhibitor that also inhibits inflammatory cytokines and disrupts inflammatory cascades
AMBP (alpha 1-Microglobulin) Transport proteins and plays a role in the regulation of inflammatory processes
B2M (beta 2-Microglobulin) Transmembrance receptor found in association with the histocompatibility complex
C9 (complement component 9) Final component of the complement system
MMP9 (matrix metallopeptidase 9) A peptidase with pro-inflammatory/anti-inflammatory activities
S100A8 (S100 calcium binding protein A8)Involved in the regulation of a number of cellular processes such as cell cycle progression and differentiation
SLPI (Secretory leukocyte protease inhibitor)A secreted inhibitor, which protects epithelial tissues from attack by endogenous proteolytic enzymes
Table 1. Selected CCBMs and their functions
AcknowledgementsSupported by the Pilot Research Award from the Sjӧgren Syndrome Foundation and the University of Iowa
College of Dentistry Dental Student Research Program College of Dentistry, University of Iowa; Paula M. Gomez Hernandez, Fang Qian, Erliang Zeng, Kim A. Brogden,
Emily A. LanzelCarver College of Medicine, University of Iowa; Scott M. Lieberman and Andrew B. Davis
University of Iowa Tissue Procurement Core
Statistical Analysis
•One way ANOVA with post‐hoc Tukey‐Kramer test
•Nonpapmetric Wilcoxon rank‐sum test
•Kruskal‐Wallis text with post‐hoc Bonferroni adjustment test
•P‐value <0.05 was used as criteria for statistical significance and 0.05<p<0.20 was used for marginal significance
Determine CCBM concentrations
•7 CCBMs selected (A2M, AMBP, B2M, C9, MMP9, S100A8, and SLPI) based on their roles in immune reactions and high concentrations in previous study
•Multiplex fluorescent bead‐based immunoassays: Luminex xMAP
Collect and prepare saliva
samples
•11 children diagnosed with and managing Sjögren Syndrome
•16 gender and age matched healthy controls
Figure 2. B2M, MMP9, and S100A8 concentrations between subjects with and without dry eye symptoms.
Figure 3. MMP9 concentrations between subjects with and without dry mouth symptoms
Figure 4. MMP9 concentrations among 3 groups of subjects with different levels of focal sialadenitis
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