P Effect of mast cell granules on the gene expression of...

Effect of mast cell granules on thegene expression of nitric oxidesynthase and tumour necrosisfactor-a in macrophages

Y. Li, T. D. Nguyen, A. C. Stechschulte,D. J. Stechschulte and K. N. DileepanCA

Division of Allergy, Clinical Immunology andRheumatology, Department of Medicine, Universityof Kansas Medical Center, 3901 Rainbow Boulevard,Kansas City, KS 66160, USA

CACorresponding AuthorTe l: (+1) 913 588 3818Fax : (+1) 913 588 3987

PREVIOUS studie s have show n that m ast cell granule s(MCG) inh ibit num erous m acrophage functionsin cluding tum our cytotox ic ity, superox ide and nitricox ide (NO) production , and FCg 2a receptor-m ediatedphagocytosis . In th is s tudy, the e ffe ct of MCG onm acrophage TNFa and n itr ic ox ide synth ase (iNOS)m RNA ex press ion , and the production and fate ofTNF a w ere ex am in ed. Upon activation w ith LPS+IFNg ,m acrophage s ex pre ssed both TNF a and iNOS m RNAand produced both TNF a and NO. Co-incubation ofLPS+IFNg -activated m acrophage s w ith MCG re sultedin dose -dependen t in hibition of iNOS m RNA ex pres -s ion . TNF a production in the activated m acrophage sw as decreased by MCG, wh ich was associated w ith areduction in TNF a m RNA ex pre ss ion . MCG w ere alsocapable of degrading both m acrophage-generated andrecom binant TNF a . Th e direct e ffect of MCG on TNF aw as partially revers ed by a m ix ture of proteasein hibitor s . These re sults dem onstrate that MCGdecrease th e production of NO and TNF a by in hibitingm acrophage iNOS and TNF a gene ex pre ss ion . Fur -therm ore, MCG post-trans crip tionally alter TNF a lev -els via proteolytic degradation .

Key words: Nitric ox ide, TNFa , Macrophage s, Mast cellgranule s, Protease s

Introduction

Macrophages play a major role in microbicidal andtumoricidal ac tivity, antigen presentation, and theproduction of a varie ty of cytokines and inflammatorymediators.1,2 Mast cells, on the othe r hand, play a keyrole in hype rsensitivity re ac tions by se creting media-tors such as histamine, proteoglycans, various cyto-kines, metabolite s of arachidonate and unique pro-te ases.3 – 6 The re cruitment of macrophages to s ites ofmast c ell degranulation and the subsequent phagocy-tosis of granule s in vivo w as first reported byFaw ce tt.7 We have pre viously show n that mast ce llgranule s (MCG) inte rac t w ith rodent macrophage sand dow nregulate superox ide production,8 –1 0

tumour cell killing and NO production.11 Thesereports confirm that mast c ell-macrophage commu-nication has a regulatory effect on host defenc emechanisms and inflammation. A role for mast c ells intumour grow th and metastasis is evident by thepresenc e of an inc re ased number of mast ce lls at theperiphery of certain tumours12 –1 4 and by the angio-genic ac tivity of seve ral mast ce ll products.1 5,1 6

Similarly, mac rophage products such as NO and TNFaare implicated in tumour grow th and inflammation.The fac t that increased number of mast c ells are foundin the periphery of tumours and MCG interactioninhibited mac rophage-mediated tumour cell killingprompte d us to ex amine the mechanism of the effe ct

of MCG on mac rophage NO and TNFa production.The focus of this study w as, there fore , to ex amine theeffect of MCG on mRNA ex pression of iNOS and TNFain macrophages. The re sults demonstrate that MCGinte rac t w ith macrophage s, inhibit iNOS and TNFamRNA ex press ion, and degrade TNFa .

Materials and methodsMaterials

Lipopolysaccharide (LPS) from Es che rich ia co li ce llw all, penicillin, streptomyc in, HEPES, metrizamide,leupeptin, pepstatin A, aprotinin, phenylmethylsulfo-nyl fluoride (PMSF) and thiazolyl blue dye (MTT) w erepurchased from Sigma Chemical Co. (St Louis , MO).Soybean trypsin inhibitor w as purchased fromBoehringe r Mannhe im (Indianapolis, IN). Heparinsolution w as supplied by Elkins-Sinn, Inc. (Cherry Hill,NJ). Minimum e ssential medium w ith Earle ’s salts(MEM) and fetal bovine serum were obtained fromHyclone Laboratorie s (Logan, UT). The ELISA kit formouse TNFa w as purchased from Genzyme (Cam-bridge , MA). Plaste k M cell culture plates w ere fromMat Tek Co. (Ashland, MA). The RNA Stat-60 kit w aspurchased from Te l-Test, Inc. (Friendswood, TX). Alltissue culture reagents used in this study w ere freefrom dete ctable leve ls of endotox in (<25 pg/ml) w hente ste d by the limulus amoebocyte lysate gel-clot assay(Associates of Cape Cod, Inc., Woods Hole , MA).

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Research Paper

Mediators of Inflammation, 7, 355–361 (1998)

Animals

Male Sprague -Daw ley rats (350–400 g) used for har-vesting mast ce lls , and male C57BL/6J mice (2–3months old), the source of macrophages, w erepurchased from Harlan Co., Indianapolis, Indiana .

Harvesting and culture of murine peritonealmacrophages

Three days prior to harve sting peritoneal ex udates,each mouse w as inje cte d intraperitone ally w ith 1.5 mlof a sterile solution of prote ose peptone (10% w /v).The pe ritoneal cavity of each mouse w as lavagedtw ice w ith 2.5 ml of minimum essential mediumcontaining 15 mM HEPES, 100 units ml of penic illin,100 m g/ml streptomyc in, 10% fetal bovine se rum(HMEM) and 5 units/ml of heparin. The pooled ce llsuspension w as sedimented by centrifugation at 2503 g for 10 min, w ashed tw ice and resuspended inHMEM w ithout heparin. Aliquots of the ce ll suspen-sion containing 2 3 105 cells w ere seeded in eachw ell of a 96-w ell culture plate . Non-adhe rent cellsw ere removed by w ashing afte r 2–4 h of incubation at37°C. Mac rophages w ere activated w ith LPS (100 ng/ml) + IFNg (10 units/ml) in all ex periments, since thisconcentrations have been found to induce optimumactivation.

Isolation of mast cells

The method employed for the isolation of mast cellshas be en pre viously described.11 ,1 7 In brief, mast c ellsw ere colle cted by lavage of the pe ritoneal andthoracic cavities of adult rats w ith 50 ml HMEMcontaining 5 units/ml heparin. The lavaged ce lls fromall animals w ere pooled, c entrifuged at 250 3 g for10 min at room temperature , and w ashed tw ice w ithHMEM. Tw o ml of the ce ll suspension containin g 6–83 107 cells we re gently laye re d on a 3 ml cushion of22.5% (w /v) metrizamide (dens ity 1.125 g/ml) inHMEM in a 15 ml centrifuge tube, and centrifuged at200 3 g for 15 min at room temperature. Mast cellsw ere sedimented at the bottom of the conical tubew hile other cells (pre dominantly macrophage s) col-lec ted at the inte rface . The mast c ell frac tions w erecollected, washed tw ice , and resuspended in HMEMw ithout heparin. Purity and viability of mast c ellsisolated by this procedure ex ceeded 95%.

Preparation of MCG and MCG sonicate

Under sterile conditions at 0–4°C, MCG were pre-pared from metrizamide-purified mast ce lls by con-trolled sonication and suc rose gradient centrifuga-tion. Mast cells w ere suspended in 1 ml of HMEM andsonicated tw ice for 20 s w ith a microtip sonicator(Sonifier Cell Disruptor, model W140) at a pow erse tting of 2.5 and temperature of 4°C.9,1 8 The

disrupted cells w ere incubated at 30°C for 15 min andmix ed vigorously for 1 min. The sonicate w as layere don 2 ml of 0.34 M sucrose and centrifuged at 50 3 gfor 10 min at 4°C. The granules at the interface w erecollected and sedimented by centrifugation at 1800 3g for 20 min at 4°C. The re sulting pellet consisting ofa homogene ous preparation of MCG w as w ashedtw ice and resuspended in the culture medium. Therecovery of granule s isolated by this procedureranged from 60% to 80% based on the his taminecontent of the starting mast cells.

MCG-sonicate w as prepared by sonicating purifiedMCG in HMEM thre e times for 30 s at max imumpow er. The quantity of MCG and MCG-sonicate usedin each ex pe riment w as ex pre ssed as the equivalentof the starting mast c ell number.

Assay of TNFa

The concentration of TNFa in the culture mediumw as assayed by both ELISA and bioassay. The sens itiv-ity of ELISA is 31 pg/ml using the protocol re com-mended by the manufacture r. For the bioassay, thetarge t L929 cells in RPMI-1640 medium w ere seeded(4.2 3 104 per w ell) in a 96-we ll culture plate . After18 h of culture, medium w as replaced w ith 5 m g/ml ofdac tinomyc in in RPMI. Tw o to 4 h after incubationw ith dac tinomyc in, se rially diluted sample s and TNFastandards (2–250 pg/ml) w ere added and incubatedfor an additional 18–24 h. The medium w as thenremoved, and 1 mg/ml of MTT dye in RPMI (w ithoutse rum and phenol red) w as added. After 2–3 h ofincubation, the medium w as aspirated comple te ly,and 100 m l of 2-propanol w as added to each w ell. Theplate w as agitated for 10 min on an orbital shaker andread on a mic rotitre plate reade r at 595 nm.

Analysis of iNOS and TNFa mRNA expression

Mouse peritone al c ells w hich contained >85% mac ro-phage s w ere seeded in a 10 cm Petri dish andincubated for 4 h to allow for macrophage adherenc e.After w ashing three times w ith HMEM, the ce lls we retre ated w ith MCG and activated w ith LPS + IFNg . Atse le cted time points afte r activation, culture mediumw as re moved, and the total cellular RNA w as ex tracte dusing an RNA ex trac tion kit. Total RNA (10 m g) w aselectrophoresed on 0.8% agarose-formaldehyde geland then transferred to Nytran nylon membrane. After2 h of pre hybridization, the membrane w as hybridizedw ith 3 2P-labelled cDNA probe specific for murineiNOS, TNFa or b -actin. The probes w ere labe lled byrandom hex amer priming method using [ a -32P] dCTPas pre viously described.19 The membrane w as thenw ashed four times and autoradiographed on a KodakX-OMAT-AR film. A scanner w as used to dete rmine thedensity of the mRNA band and the re lative density ofeach band w as normalized to b -actin.

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356 Mediators of Inflammation · Vol 7 · 1998

Treatment of MCG with protease inhibitors

MCG w ere incubated w ith a mix ture of proteaseinhibitor s containing PMSF 2.5 mM, pepstatin A5 m g/ml, trypsin inhibitor 50 m g/ml, leupeptin50 m g/ml, and aprotinin 50 m g/ml, at 37°C for 2 h. Theprotease inhibitor-treated MCG w ere then evaluatedfor the ir e ffec ts on macrophages.

Statistical analysis

Whenever applicable, the data we re analysed by one-w ay analysis of variance w ith subsequent Student-New man-Kue ls ’ te st. All re sults w ere ex pressed asmeans ± SEM and P<0.05 w as conside red significant.

ResultsEffects of MCG on macrophage iNOS mRNAexpression

Earlie r w ork demonstrated that MCG inhibit macro-phage NO production.11 To inve stigate the mecha-nism, the effe ct of MCG on macrophage iNOS mRNAex pre ssion w as analysed at se le cted mast ce ll-to-macrophage ratios. The addition of MCG simultane-ously w ith LPS+IFNg to macrophage monolayerinhibited iNOS mRNA ex pression in a dose-dependent

manner (Fig. 1). Max imal inhibition w as noted at aMCG dose equivalent to a mast c ell-to-macrophageratio of 1:5 and the inhibition w as evident even at aratio of 1:40. When macrophages w ere pre-incubatedw ith MCG for 3 h prior to activation, the inhibitoryeffect of MCG decreased compared w ith simultane-ous addition of MCG and the ac tivators. Furthermore ,if macrophages we re incubated w ith MCG for morethan 6 h prior to ac tivation, there w as no inhibition ofthe ex pression of iNOS transcript (Fig. 2).

Effects of MCG on TNFa production

When ac tivated w ith LPS+IFNg , macrophages gen-erated large amounts of TNFa . Addition of MCG orMCG-S at the time of LPS+IFNg stimulation resulted indose-dependent decre ase in TNFa le ve ls and morethan a 90% decrease w as note d at a MCG doseequivalent to a mast c ell-to-macrophage ratio of 1.5 to2 (Fig. 3). The MCG-induced decrease in TNFa leve lsw as se en w hen determined by ELISA (Fig. 3A) and bybioassay (Fig. 3B). Although the assayed value s dif-fered for the same sample s depending on the assaymethod employed, the decrease of TNFa le ve ls wasevident. There w as no statistic ally significant differ-ence be tw een the effe cts of MCG and MCG-S.Therefore , in later ex pe riments only the intact MCGw ere used. Unactivated macrophages or those

Re gu la tio n o f m a cro phage NO a nd TNFa pro du ctio n by m a s t ce lls

Mediators of Inflammation · Vol 7 · 1998 357

1 2 3 4 5 6

FIG. 1. Effect of MCG on macrophage iNOS mRNA expres-sion. Adherent macrophages (1 3 107) were exposed to theindicated doses of MCG (mast cell equivalent) and activatedwith LPS (100ng/ml) + IFNg (10units/ml). In all subsequentexperiments the macrophages were activated with theseconcentrations. Eighteen hours after activation, iNOS andb -actin mRNA were analysed by Northern blotting. Theb -actin expression was used to normalize the data. For toppanel: lane 1. control; lane 2. LPS+IFNg ; lane 3. LPS+IFNg+MCG 23 106; lane 4. LPS+IFNg +MCG 13 106; lane 5.LPS+FNg +MCG 0.53 106; lane 6. LPS+IFNg +MCG 0.253 106.This is representative of three experiments.

FIG. 2. Effect of pre-treatment of macrophages with MCG oniNOS mRNA expression. Macrophages (1 3 107) werecultured with MCG (2 3 106 mast cell equivalent) for 24, 6and 3h prior to activation or added simultaneously with LPS+ IFNg . Eighteen hours after activation, iNOS and b -actinmRNA were analysed by Northern blotting. The b -actinexpression was used to normalize the data. This is repre-sentative of three experiments.

ex posed only to MCG or MCG-S did not producedete ctable leve ls of TNFa .

The production of TNFa by mac rophages w asevident as early as 1.5 h afte r activation. The cumu-lative TNFa leve l inc reased for up to 24 h w hile therate of production appeared to decrease (Fig. 4). Thepresenc e of MCG decre ased the leve l of TNFa byapprox imately 40% at 1.5 and 6 h in contrast to a 95%decre ase at 24 h. This e ffec t w as also evident w henTNFa w as as sayed by bioassay (data not show n).

Effects of MCG on TNFa mRNA expression

In order to as sess the e ffec t of MCG on TNFa mRNAex pre ssion by ac tivated macrophages, the transc riptsw ere analysed at 2, 6 and 24 h of incubation afteractivation. The result demonstrate s that TNFa mRNAw as rapidly ex pre ssed to a max imal leve l w ithin 2 hafter activation and then dec lined the reafter (Fig. 5).Addition of MCG to LPS+IFNg -stimulated mac ro-phage s inhibited TNFa mRNA ex press ion by 60% at amast c ell-to-macrophage ratio of 1:3.

Effect of MCG proteases on TNFa degradation

Rat mast c ell granules contain a varie ty of prote asesincluding tryptase, chymase , and carbox ypepti-

Y. Li e t al.


FIG. 3. Dose response effect of MCG or MCG-S on macro-phage TNFa production. Adherent macrophages (200,000)were cultured with LPS + IFNg in the presence or absence ofthe indicated doses of MCG or MCG-S (mast cell equivalent).After 6h of culture, media were collected and TNFa levelswere measured by ELISA (panel A) and bio-assay (panel B)utilizing L929 cells as the target. Each value presented is themean ± SEM of quadruplicate determinations. There is nosignificant difference between the effects of MCG and MCG-S. This is representative of three experiments.*P<0.05 vs. noMCG added.

FIG. 4. Time course of the effect of MCG on macrophagesecreted TNFa levels. Adherent macrophages (200,000) werecultured with LPS + IFNg in the presence or absence of MCG(100000 mast cell equivalent). After the indicated times, mediawere collected and TNFa levels were measured by ELISA.Each value presented is the mean ± SEM of quadruplicatedeterminations. This is representative of two experi-ments.*P<0.05 vs. LPS+IFNg only at the same time point.

FIG. 5. Effect of MCG on macrophage TNFa mRNA expres-sion. Macrophages (1.5 3 107) were cultured without or withMCG (5 3 106 mast cell equivalent) and simultaneouslyactivated with LPS + IFNg . After the indicated culture times,TNFa and b -actin mRNA were analysed by Northern blotting.The b -actin expression was used to normalize the data. Thisis representative of three experiments.

dase .20 – 2 3 To inve stigate if TNFa is susceptible tothese enzymes, conditioned media w ith know n leve lsof TNFa from LPS+IFNg -ac tivated macrophage s orrecombinant mouse TNFa were incubated w ith MCGfor 24 h at 37°C. The TNFa le ve ls, assayed by ELISA,indicate that both recombinant and mac rophage-se creted TNFa were degraded by MCG (Fig. 6). Thiseffect w as partially re versed by the pre -treatment ofMCG w ith a mix ture of protease inhibitors containing

PMSF, pepstatin A, trypsin inhibitor, leupeptin andaprotinin (Fig. 7). The complete abrogation of TNFadegradation by MCG could not be achieved even afterex tending the pre-treatment of MCG w ith the seprotease inhibitors to 24 h at 4°C.

DiscussionThe pre sent study demonstrates that incubation ofmacrophage s w ith MCG during activation w ithLPS+INFg results in the inhibition of iNOS and TNFamRNA ex pre ssion w ith a consequent dec rease in NOand TNFa production. This corroborates our previousreport documenting MCG inhibition of NO produc-tion and tumour cell killing by LPS+INFg -activate dmacrophage s.11 The MCG effe ct was evident even atan estimated mast ce ll to macrophage ratio of 1:40, are lationship like ly to occur in v ivo . The inhibitoryeffect of MCG on macrophage iNOS mRNA ex pre s-sion seems transient be cause this e ffec t disappearsw hen the macrophages w ere pre -incubated w ithMCG for 6 h or more . The same phenomenon wasobserved w ith the inhibito ry effect of MCG on TNFamRNA ex pression (data not show n). Our earlierstudies have ruled out the possibilities that MCG mayinactivate LPS or IFNg . Further inve stigation is neededto clarify the mechanism by w hich MCG inhibitmacrophage iNOS and TNFa mRNA ex press ion in atransient manner.

The ex pression of TNFa by activated mac rophagesis of particular significance since the gene s encodingiNOS and TNFa can be coordinate ly regulated. In v ivostudies have suggested a role for TNFa in the positiveregulation of iNOS follow ing LPS injec tion. Anti-TNFaantibodie s or soluble TNFa receptor antagonistspartially block LPS-induced pulmonary iNOS activityor hepatic iNOS mRNA ex pression re spec tively.2 4,2 5

Furthe rmore , the involvement of TNFa in both theinduction and maintenance of iNOS mRNA in mac ro-phage s w as re cently reported using an in v ivo mousemode l.26 These studies indicate that TNFa is anautocrine regulator of iNOS ex pre ssion. The TNFamRNA ex pre ssion begins as early as 0.5 h,2 7 w hereasiNOS mRNA cannot be de tec te d until 8 h afteractivation.28 Our data also show that TNFa mRNAex pre ssion by macrophages is optimal at 2 h afteractivation and then progressive ly dec reases . Based onthose data w e conclude that the depletion of mac ro-phage -se creted TNFa by MCG could contribute toinhibition of LPS-induced iNOS mRNA ex pression. Itis clear that the MCG to macrophage ratio thatcomple te ly inhibits the ex pression of iNOS mRNAonly partially inhibits the ex press ion of TNFa mRNA.We do not yet have an ex planation for this phenom-enon. It is poss ible that the iNOS ex press ion needs afinite amount of TNFa to trigge r the gene .

Rodent mast ce lls contain histamine, serotonin,and prote ases including serine proteases, neutral



FIG. 6. TNFa degradation by MCG. Macrophage derived andrecombinant TNFa were incubated with MCG (100,000 mastcell equivalent) in a volume of 0.2ml culture medium for24h. TNFa was assayed by ELISA. Values presented are themean ± SEM of quadruplicate determinations. This is arepresentative of four experiments.*P<0.05 vs. no MCG.

FIG. 7. Effect of protease inhibitors on TNFa degradation byMCG. Mouse recombinant TNFa was incubated with theindicated doses of MCG in cell culture medium for 24h. MCGwere incubated with the protease inhibitor mixture at 37°Cfor 2h before being added to TNFa . The TNFa levels wereassayed by ELISA. Values presented are the mean ± SEM ofquadruplicate determinations. The protease inhibitor mix-ture contains 2.5mM PMSF, 5 m g/ml pepstatin A, 50 m g/mleach of trypsin inhibitor, leupeptin and aprotinin. This isrepresentative of three experiments.*P<0.05 vs. control.

protease s, and carbox ypeptidase A.2 0 – 23 Histamineand serotonin at concentrations present in mastce lls (20 m g and 2 m g per million cells re spectively)may induce immunomodulatory e ffec ts. Forinstance, histamine up-regulates macrophage synthe-sis of IL-1.29 However, histamine at concentrationsin the range of 10– 6 –10– 3 M (equalling and ex ceed-ing that present in MCG used in this study) failedto affe ct TNFa and NO production by mac rophages(data not show n). The se results are in agreementw ith our earlier re port of MCG inhibition of macro-phage superox ide production, w hich show ed thatunlike MCG, histamine and se rotonin w ere ineffe c-tive in modulating the respiratory burst.9 The e ffec tof serotonin on macrophage NO and TNFa w as notte ste d in this study. It is also notew orthy thathistamine and serotonin are short-lived and theireffects are rapidly lost afte r mast cell degranulation.Heparin is an important constituent of MCG andmurine mac rophages possess heparin receptors .30 Aprevious report has show n that commercial heparinand MCG are capable of inhibiting Fc g 2a receptormediated phagocytosis in macrophage cell line.3 1

Although commercial heparin differs from rat MCG-heparin, it is possible that MCG-heparin may havesome modulatory e ffec ts on macrophage iNOS andTNFa ex press ion. Our results presented here showthat both MCG and MCG-sonicate are capable ofdegrading TNFa w ith loss of immunoreactivity andbiological activity, and that both se cre ted andrecombinant TNFa are susceptible . This indicatesthat the decre ased leve ls of TNFa could be partiallydue to the prote olytic effe cts of MCG-proteases, inaddition to the effect of MCG inhibition of macro-phage TNFa mRNA ex pre ssion. However, this pro-te olytic effe ct w as only partially re ve rsed by treat-ment of MCG w ith a mix ture of protease inhibitors.The failure of protease inhibitors to comple te lyabrogate the MCG effe ct may be due to incomple teinhibition of the protease activity or due to theabsence of specific inhibitor s for certainprotease s.

Mast ce ll degranulation gene rates the re lease of avarie ty of inflammatory molecules that dire ctly stim-ulate many ce ll types including epithelial cells,3 2

fibroblasts33 and endothelial ce lls .17 This study is thefirst e vidence that mast c ell granule s inhibit macro-phage TNFa and iNOS mRNA ex pre ssion. We alsofound that the mast cell proteases degrade recombi-nant and macrophage sec rete d TNFa . The pre - andpost-transcriptional regulation of TNFa production inLPS-activated macrophages by MCG may contribute tothe inhibition of iNOS mRNA ex press ion.

References1. Adams DO, Hamilton TA. The ce ll biology of mac rophage activation. An n

Re v Im m u n o l 1984; 2: 283–318.

2. Adams DO, Hamilton TA. Molecular bas is of mac rophage activation. In:Lew is CE, McGee JO’D, e ds . Th e Mac ro pha g e s . New York: Ox fordUnive rsity Pres s, 1992; 75–114.

3. Ishizaka T, Ishizaka K. Activation of mast c e lls for mediator re leasethrough IgE rece ptors. Pro g Alle rg y 1984; 34: 188 –235.

4. Schw artz LB, Austen KF. Structure and function of the chemicalmediators of mast ce lls . Pro g Alle rg y 1984; 34: 271–321.

5. Galli SJ, Lichte nste in LM. Biology of mast ce lls and basophils. In:Middle ton E Jr, ed. Alle rg y : princ iple s a n d pra c tice s , 3rd edn. St Louis,MO: Mosby, 1988; 160.

6. Holgate ST, Robinson C, Church MK. Mediators of immediate hyper-se nsitivity. In: Middle ton E Jr, ed. Alle rg y : pr in cip le s a n d pra c tice s , 3rdedn. St Louis, MO: Mosby, 1988; 135.

7. Faw cett DW. An ex perimental s tudy of mast ce ll de granulation andre gene ration. An a t Re c 1955; 121 : 29–43.

8. Dileepan KN, Stechschulte DJ. Influence of mast ce lls on mac rophagefunction. Bio ch em So c Tra n s 1986; 14: 913–914.

9. Dileepan KN, Simpson KM, Stechschulte DJ. Modulation of mac rophagesuperox ide-induced cytochrome c reduction by mast ce lls . J La b ClinMed 1989; 113 : 577–585.

10. Shah BA, Li Y, Stechschulte DJ, Dile epan KN. Phagocytos is of mast c e llgranules re sults in dec reased macrophage supe rox ide production. MedIn flam m 1995; 4: 406 –412.

11. Dileepan KN, Lorsbach R, Stechschulte DJ. Mast ce ll granules inhibitmacrophage-me diate d lysis of mastocytoma ce lls (P815) and nitric ox ideproduction. J Le uko c Bio l 1993; 53: 446–453.

12. Caw ley EP, Hoch-Ligeti C. Assoc iation of tis sue mast c e lls and skintumors . Ar ch De rm a to l 1961; 83: 146–150.

13. Bow e rs HM, Mahapatro RC, Ke nnedy JW. Numbers of mast ce lls in theax illary lymph nodes of breast canc er patie nts. Ca nc e r 1979; 43:568–573.

14. Roche WR. Mast c e lls and tumors. The spec ific e nhanc ement of tumorprolife ration in v itro. Am J Pa tho l 1985; 119 : 57–64.

15. Qu Z, Liebler JM, Pow ers MR, Gale y T, Ahmadi P, Huang XN, Anse l JC,Butte rfie ld JH, Planck SR, Rosenbaum JT. Mast ce lls are a major source ofbasic fibroblast grow th factor in chronic inf lammation and c utaneoushe mangioma. Am J Pa tho l 1995; 147 : 564–573.

16. Kaartinen M, Penttila A, Kovanen PT. Mast c e lls accompany micro-vesse ls in human c oronary athe romas: implic ations for intimal ne o-vasc ularization and hemorrhage . Athe ro s c le ro s is . 1996; 123 :123–131.

17. Li Y, Stechschulte AC, Smith DD, Lindsle y HB, Stechschulte DJ, Dilee panKN. Mast ce ll granules potentiate endotox in-induce d inte rleukin-6production by endothe lial c e lls. J Le uko c Bio l 1997; 62: 210–216.

18. Raphael GD, Henderson WR, Kaliner M. Isolation of me mbrane-bound ratmast c e ll granules. Ex p Ce ll Re s 1978; 115 : 428–431.

19. Li Y, Ito N, Suzuki T, Ste chschulte DJ, Dile epan KN. Dex amethasoneinhibits nitric ox ide -mediated cytotox ic ity via e ffe ct on both macro-phage s and targe t ce lls . Im m u n o pha rm a co lo g y 1995; 30: 177–186.

20. Yurt R, Austen KF. Preparative purific ation of the rat mast ce ll chymase :characte r ization and interac tion w ith granule components . J Exp Med1977; 146 : 1405–1419.

21. Woodbury RG, Ever itt MT, Ne urath H. Mast c e ll proteases. MethEnzy m o l 1981; 80: 588–609.

22. Kokkonen JO, Vartiaine n M, Kovanen PT. Low dens ity lipoprote indegradation by sec retory granules of rat mast ce lls . Sequential degrada-tion of apolipoprote in B by granule chymase and carbox ypeptidase A. JBio l Chem 1986; 261 : 16067–16072.

23. Schw artz LB, Irani AM, Rolle r K, Caste lls MC, Schechte r NM. Quantita-tion of histamine , tryptase and chymase in dispe rse d human T and TCmast c e lls. J Im m u n o l 1987; 138 : 2611–2615.

24. Thiemermann C, Wu CC, Szabo C, Perre tti M, Vane JR. Role of tumournec rosis fac tor in the induc tion of nitric ox ide synthase in a rat model ofendotox in shock. Br J Pha rm a co l 1993; 110 : 177–182.

25. Ge lle r DA, de Vera ME, Russe ll DA, Shapiro RA, Nussle r AK, Simmons RL,Billiar TR. A central role for IL-1 b in the in vitro and in v ivo re gulation ofhe patic inducible nitric ox ide synthase . J Im m u n o l 1995; 155 :4890–4898.

26. Salkow ski CA, De tore G, McNally R, van-Rooijen N, Voge l SN. Regulationof inducible nitric ox ide synthase messenger RNA ex pres sion and nitricox ide produc tion by lipopolysaccharide in vivo: the role s of macro-phage s, endogenous IFNg , and TNF receptor-1-mediated signaling. JIm m u no l 1997; 158 : 905–912.

27. Marchant A, Gue ydan C, Houzet L, Amraoui Z, Se ls A, Huez G, GoldmanM, Kruys V. De fective translation of tumor necros is factor mRNA inlipopolysaccharide-tole rant macrophages . J In fla m m 1996; 46:114–123.

28. Lorsbach RB, Murphy WJ, Low enste in CJ, Snyde r SH, Russe ll SW.Ex press ion of the nitric ox ide synthase gene in mouse macrophagesactivated for tumor ce ll killing. J Bio l Ch em 1993; 268 : 1908 –1913.

29. Okamoto H, Nakano K. Regulation of interle ukin-1 synthes is byhistamine produce d by mouse peritone al mac rophage s pe r se . Im m u -n o lo g y 1990; 69 : 162–165.

30. Ble ibe rg I, MacGregor I, Aronson M. Heparin rec eptors on mousemacrophages . Th ro m b Re s 1983; 29: 53–61.

31. Yamada A, Dileepan KN, Ste chschulte DJ, Suzuki T. Regulation of Fc g 2are ceptor-mediated phagocytosis by a murine macrophage like ce ll line ,

Y. Li e t al.


P388D1. Involvement of c ase in kinase II activity assoc iated w ith Fc g 2are ceptor. J Mo l Ce ll Im m u n o l 1989; 4: 191–202.

32. Cairns JA, Walls AF. Mast c e ll tryptase is a mitogen for epithe lial ce lls .Stimulation of IL-8 production and interce llular adhes ion molecule -1ex press ion. J Im m un o l 1996; 156 : 275–283.

33. Gruber BL, Kew RR, Je laska A, Marchese MJ, Garlick J, Ren S, Schw artz LB,Korn JH. Human mast ce lls activate fibroblasts. Tryptase is a fibroge nicfac tor s timulating collagen messenger ribonucle ic acid synthe sis andfibroblast chemotax is. J Im m u no l 1997; 158 : 2310–2317.

ACKNOWLEDGEMENTS. This s tudy w as supported by grants from theAmerican He art Assoc iation (Kansas affiliate ) and the Univers ity of KansasMe dical Center Re search Institute , and by the Joseph and Elizabe th Care yArthritis Funds , Hinman Fund, and Jones Fund from the KU Endow mentAssoc iation.

Received 16 August 1998accepted 1 September 1998



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