IL-4 Is an Endogenous Inhibitor of Neutrophil Influx and Subsequent Pathology in Acute Antibody-Mediated Inflammation This information is current as Sohail Saleem, Zhenhua Dai, Sandra N. Coelho, Bogumila T. of October 3, 2021. Konieczny, Karel J. M. Assmann, Fady K. Baddoura and Fadi G. Lakkis J Immunol 1998; 160:979-984; ; http://www.jimmunol.org/content/160/2/979 Downloaded from References This article cites 51 articles, 21 of which you can access for free at: http://www.jimmunol.org/content/160/2/979.full#ref-list-1 Why The JI? Submit online. http://www.jimmunol.org/ • Rapid Reviews! 30 days* from submission to initial decision • No Triage! Every submission reviewed by practicing scientists • Fast Publication! 4 weeks from acceptance to publication *average by guest on October 3, 2021 Subscription Information about subscribing to The Journal of Immunology is online at: http://jimmunol.org/subscription Permissions Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html Email Alerts Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts The Journal of Immunology is published twice each month by The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 1998 by The American Association of Immunologists All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. IL-4 Is an Endogenous Inhibitor of Neutrophil Influx and Subsequent Pathology in Acute Antibody-Mediated Inflammation1 Sohail Saleem,* Zhenhua Dai,* Sandra N. Coelho,† Bogumila T. Konieczny,* Karel J. M. Assmann,‡ Fady K. Baddoura,§ and Fadi G. Lakkis2* IL-4 is an immunoregulatory cytokine that has in vitro and in vivo anti-inflammatory actions. In this study we investigated whether endogenously produced IL-4 modulates inflammatory processes that occur after Abs bind to target tissue by comparing the severity of glomerulonephritis induced by heterologous anti-glomerular basement membrane Abs in wild-type (IL-41/1) mice to that of glomerulonephritis induced in homozygous IL-4 gene knockout (IL-42/2) mice. Two hours after Ab injection, 2/2 IL-4 mice had significantly higher intrarenal intercellular adhesion molecule-1 mRNA expression and intraglomerular neu- Downloaded from trophil accumulation than the IL-41/1 group. Treatment of IL-42/2 mice with recombinant murine IL-4 at the time of disease induction reduced intercellular adhesion molecule-1 expression and neutrophil influx to levels observed in IL-41/1 kidneys. Four days after Ab administration, untreated IL-42/2 mice developed significantly greater urinary protein excretion, intracap- illary fibrinogen deposits, and glomerular hypercellularity than IL-41/1 mice. These results demonstrate that endogenous IL-4 suppresses neutrophil influx and limits tissue damage in Ab-induced glomerulonephritis, suggesting that IL-4 is an important regulator of acute inflammatory processes. The Journal of Immunology, 1998, 160: 979–984. http://www.jimmunol.org/ L-4 is an immunoregulatory cytokine produced by Th2 lym- press greater amounts of IL-4 are less susceptible (17, 18). San- phocytes, mast cells, basophils, and a subset of NK cells (1, tiago et al. demonstrated that IL-4 overexpression in lupus-prone I 2). Although IL-4 induces IgE synthesis and promotes im- mice prevents glomerulonephritis by down-regulating Th1-driven mediate-type hypersensitivity reactions, there is evidence to sug- production of complement-fixing, nephrotoxic subclasses of IgG gest that IL-4 has in vitro and in vivo anti-inflammatory actions (1, autoantibodies (19). It is not known, however, whether IL-4 mod- 2). IL-4 inhibits IFN-g production (3, 4) and suppresses IFN-g- ulates inflammatory processes that occur after Abs deposit in renal mediated macrophage activation (5, 6). IL-4 also induces differ- tissue. These processes include leukocyte migration and activation, entiation of naive T cells to Th2 lymphocytes (7, 8) that secrete proteinuria, fibrin accumulation, and, eventually, glomerular by guest on October 3, 2021 additional macrophage-inhibiting cytokines such as IL-10 and fibrosis (16). IL-13 (9, 10). Other anti-inflammatory actions of IL-4 include sup- In this study we examined whether IL-4 is an endogenous in- pressing IL-8 production by neutrophils (11) and reducing proco- hibitor of acute, Ab-mediated tissue inflammation by comparing agulant activity expression by activated endothelial cells (12). In the severity of anti-glomerular basement membrane (anti-GBM)3 vivo administration of IL-4 has been shown to protect against ex- nephritis in IL-41/1 to that in IL-42/2 mice. Murine anti- perimental arthritis and immune complex-induced lung inflamma- glomerular basement membrane (anti-GBM)3 nephritis is initiated tion in rats (13, 14). Pancreatic overexpression of IL-4 also pre- by binding of heterologous (sheep, rabbit, or goat) Abs of the IgG vented insulitis in nonobese diabetic mice (15). class to mouse GBM followed by rapid neutrophil accumulation in Glomerular inflammation in humans and experimental animals glomerular tissue (20, 21). Marked proteinuria, fibrinogen depos- is often initiated by binding of Abs to target Ags in the kidney (16). its, glomerular capillary obliteration, and increased glomerular cel- The severity of Ab-mediated glomerulonephritis in rats and lupus- lularity develop within several hours to a few days after Ab bind- prone mice was found to correlate with the host’s propensity to ing (20, 21). Glomerular injury in this disease model is neutrophil mount a Th1-type immune response characterized by predominant dependent (22–24). The present report provides direct evidence IFN-g and IL-2 expression (17, 18). In contrast, animals that ex- that IL-4 is an endogenous inhibitor of neutrophil infiltration and subsequent glomerular pathology in Ab-induced nephritis. *Renal Division, Emory University School of Medicine and Veterans Affairs Med- ical Center, Atlanta, GA 30033; †Renal Division, Center of Health Sciences, Materials and Methods ‡ Federal University of Pernambuco, Recife, Brazil; Department of Pathology, Animals University of Nijmegen, Nijmegen, The Netherlands; and §Department of Pa- thology and Laboratory Medicine, Veterans Affairs Medical Center and State Six- to eight-week-old male C57BL/6 wild-type (IL-41/1) mice were pur- University of New York, Buffalo, NY 14215 chased from The Jackson Laboratory (Bar Harbor, ME). C57BL/6 mice Received for publication June 11, 1997. Accepted for publication October homozygous for the disrupted IL-4 genes (IL-42/2) were provided by Dr. 3, 1997. Manfred Kopf (Basel Institute of Immunology, Basel, Switzerland) (7) and The costs of publication of this article were defrayed in part by the payment of were bred at the Atlanta Veterans Affairs Medical Center animal facility. page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact. 3 Abbreviations used in this paper: GBM, glomerular basement membrane; NTS, 1 This work was supported by a Veterans Affairs Merit Review grant (to F.G.L.). nephrotoxic serum; NRS, nonimmune rabbit serum; Up/UCr, ratio of urinary pro- 2 Address correspondence and reprint requests to Dr. Fadi G. Lakkis, Emory tein to creatinine concentration; PAS, periodic acid-Schiff; CAE, chloroacetate University and Veterans Affairs Medical Center, Research 151N, 1670 Clairmont esterase; RT, reverse transcription; GAPDH, glyceraldehyde-3-phosphate dehy- Rd., Atlanta, GA 30033. E-mail: [email protected] drogenase; ICAM-1, intercellular adhesion molecule-1. Copyright © 1998 by The American Association of Immunologists 0022-1767/98/$02.00 980 ANTI-INFLAMMATORY ACTIONS OF ENDOGENOUS IL-4 Complete inactivation of IL-4 gene function was verified by performing IL-4-specific ELISA (Genzyme, Cambridge, MA) on splenocyte superna- tants collected 0, 24, 48, and 72 h after Con A (3 mg/ml) stimulation. Experimental protocols Expt. A. To study the effect of IL-4 on Ab-mediated neutrophil influx, anti-GBM nephritis was induced in 8- to 10-wk-old IL-41/1 and IL-42/2 C57BL/6 mice (n 5 6/group) by injecting 1 ml of heat-inactivated neph- rotoxic serum (NTS; rabbit anti-mouse GBM) i.p. (20). NTS was provided by Dr. Karel J. M. Assmann (University Hospital, Nijmegen, The Nether- lands). An additional group of IL-42/2 C57BL/6 mice (n 5 6) received FIGURE 1. RT-PCR analysis of intrarenal IL-4 mRNA expression be- 3000 U of recombinant murine IL-4 (a gift from Dr. Satwant Narula, fore (pre-NTS) and 2 h after nephrotoxic serum administration (post- Schering-Plough Research Institute, Kenilworth, NJ) i.v. at the time of 1 1 2 2 1 1 2 2 / 1 1 / 2 2 NTS administration. Control IL-4 / and IL-4 / (n 5 3/group) mice NTS). Total RNA was prepared from IL-4 ( / ) and IL-4 ( / ) received 1 ml of heat-inactivated nonimmune rabbit serum (NRS; Sigma kidneys, reverse transcribed, and subjected to 38 cycles of PCR. Fif- Chemical Co., St. Louis, MO) i.p. All mice were killed 2 h later, and renal teen percent of each PCR reaction was electrophoresed on 2% agarose tissue was saved for analysis. gels and stained with ethidium bromide. Results were similar in three Expt. B. To study the effects of IL-4 on glomerular pathology that occurs separate animal experiments. L, DNA ladder (fX174RF DNA/HaeIII subsequent to neutrophil infiltration, anti-GBM nephritis was induced in 8- fragments); C , control PCR reaction without reverse transcriptase; C , 1/1 2/2 5 1 2 to 10-wk-old IL-4 and IL-4 C57BL/6 mice (n 10/group) by in- control PCR reaction without cDNA. jecting 1 ml of heat-inactivated NTS i.p. on day 0 (20). Control IL-41/1 and IL-42/2 mice (n 5 3/group) received 1 ml of heat-inactivated NRS (Sigma) i.p.