IL 17 in Disease
IL 17 in Disease
IL 17 in Disease
REVIEW ARTICLE
Summary Interleukin-17 (IL-17) has emerged as a central player in the mammalian immune system. Although this cytokine exerts a host-defensive role in many infectious diseases, it promotes inammatory pathology in autoimmunity and other settings. A myriad of studies have focused on how IL-17-producing cells are generated. However, the means by which IL-17 achieves its effects, either for the benet or the detriment of the host, are due in large part to the induction of new gene expression. Whereas many IL-17 target genes are common to different disease states, in some cases the effects of IL-17 differ depending on the target cell, infectious site or pathogen. Gene products induced by IL-17 include cytokines (IL-6, granulocyte-colony-stimulating factor, tumour necrosis factor-a), chemokines (CXCL1, CXCL2, CCL20, among many others), inammatory effectors (acute-phase protesins, complement) and antimicrobial proteins (defensins, mucins). Different cell types appear to respond differently to IL-17 in terms of target gene expression, with notable differences seen in mesenchymal and epithelial cells compared with cells of haematopoietic origin. Here, we summarize the major IL-17 target genes that mediate this cytokines activities in both autoimmune and chronic diseases as well as during various types of infections. Keywords: cytokine; gene target; interleukin-17; inammation; signal transduction
doi:10.1111/j.1365-2567.2009.03240.x Received 19 November 2009; revised 3 December 2009; accepted 15 December 2009. Correspondence: S. L. Gaffen, University of Pittsburgh, Division of Rheumatology & Clinical Immunology, S708 Biomedical Science Tower, 3500 Terrace Street, Pittsburgh PA 15261, USA. Email: [email protected] Senior author: Sarah L. Gaffen
Introduction
The interleukin-17 (IL-17) family is the most recently described subclass of cytokines.1 Since 2000, we have started to gain an understanding of IL-17 family members and their corresponding receptors, which has led to new insights into how immunity to infections and autoimmunity are governed. To date, there are six IL-17-family ligands [IL-17A, IL-17B, IL-17C, IL-17D, IL-17E (IL-25) and IL-17F], and ve receptors (IL-17RA, IL-17RB/ IL-25R, IL-17RC, IL-17RD/SEF and IL-17RE).2 Interleu-
kin-17A (hereafter referred to as IL-17) is the most intensively studied, but interest in the rest of the family is growing. Originally IL-17 was thought to be produced exclusively by T cells,3 but it is now known to be secreted by a variety of innate cells including macrophages, dendritic cells (DC), natural killer, natural killer T, lymphoid tissue inducer and cd-T cells.4 A major development in this eld occurred with the recognition that IL-17-producing CD4+ T cells arise as a population distinct from the classic T helper type 1 (Th1) and Th2 cells.57 Whereas it was
Abbreviations: APC, antigen-presenting cell; BAFF, B-cell activating factor; BD, b-defensin; C/EBP, CCAAT/enhancer binding protein; DC, dendritic cell; DSS, dextran sulphate sodium; EAE, experimental autoimmune encephalomyelitis; GC, germinal centre; G-CSF, granulocyte colony-stimulating factor; GWAS, genome-wide association studies; IBD, inammatory bowel disease; IFN, interferon; IL, interleukin; Lcn2, lipocalin 2/24p3; MMP, matrix metalloproteinase; NF-jB, nuclear factor-jB; NOD, nucleotide oligomerization domain; RA, rheumatoid arthritis; RANKL, receptor activator of nuclear factor-jB ligand; SEFIR, SEF/IL17R; SLE, systemic lupus erythematosus; STAT5, signal transducer and activator of transcription 5 ; TGF, transforming growth factor; Th, T helper; TLR, Toll-like receptor; TMEV, Theilers murine encephalomyelitis virus; TNBS, trinitrobenzene sulphonic acid; TNF, tumour necrosis factor; VV, vaccinia virus.
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Autoimmune EAE/brain Bloodbrain CC-chemokines barrier CXC-chemokines Neuron IL-22 Th17 IL-17 GranzymeB Psoriasis/skin IL-22
Th17
LL37 S100As
IL-17 Th17 G-CSF Influenza CC-chemokines CXC-chemokines Gut mucosa C. rodentium Regllls IL-17 IL-17F Mf
Th17
S100As
IL-17
Twist1 Reg13,16
IL-17 (suppression)
Th17
T-bet
CC-chemokines CXC-chemokines
Figure 1. IL-17 signaling and target genes in various disease settings. In blue are representative mucosal infections where IL-17 plays a key role, along with key target genes involved in each. In pink are autoimmune diseases and the role of IL-17 and particular target genes therein.
Rheumatoid arthritis
Many, if not most, autoimmune diseases are now connected in some manner to IL-17 or the Th17 pathway. In particular, RA has been intensively studied, starting even before the recognition of the Th17 subset. The key features of inamed arthritic joints are proliferating synovial broblasts, joint and cartilage erosion, inltrating CD4+ T cells and autoantibody-producing plasma cells. In addition, increased numbers of innate immune cells (DC, granulocytes and macrophages), in some cases ectopic germinal centres (GC) are found within joints. Early studies showed that high levels of IL-17 were found in the rheumatoid synovium of patients with RA but not of controls or of patients with osteoarthritis. Consistently, adding IL-17 to an in vitro culture system stimulated bone resorption and collagen destruction.23 Furthermore, neutralizing IL-17 or its receptor in collagen-induced arthritis mouse models resolved RA symptoms, IL-17Adecient mice are protected from collagen-induced
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arthritis, and adding IL-17 ectopically by gene therapy exacerbated disease.2426 Consequently, IL-17 appears to promote both inammation and bone destruction in RA. These ndings pose the question of how IL-17 mediates its pathogenic activities. Interleukin-17 induces pro-inammatory cytokines such as TNF-a, IL-1b and IL-6 from cartilage, synoviocytes, macrophages and bone cells.27 Collectively, these pro-inammatory cytokines contribute to RA are-ups and also establish a chronic inammatory state by a self-reinforcing positive feedback loop wherein IL-17-induced IL-6 maintains the Th17 T-cell population.28 The IL-17 also stimulates the production of multiple chemokines, including IL-8/CXCL8, CXCL1 (KC/Groa), CXCL2 (MIP2a/Grob), CCL20 (MIP-3a), CCL2 (MCP1) and CCL7 (MCP3).3,5,27,29 These serve to recruit neutrophils, macrophages and lymphocytes to the synovium, thereby enhancing inammation. Secondary lymphoid GC formations are found ectopically in RA synovial tissue.30,31 Although it has been reported that synovial lymphoid neogenesis is not a major determinant of RA-specic autoantibody responses, this phenomenon is indicative of ongoing inammation.32
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Irreversible deformities in joints are a key feature of advanced RA, caused by extensive cartilage and bone erosion (Table 1). Interleukin-17 contributes to this process by inducing expression of matrix metalloproteinases (MMP) 1, 2, 3, 9 and 13, which drive degradation of extracellular matrix within the joint.3339 It also induces prostaglandin E2 via cyclooxgenase-2, which enhances inammation by many mechanisms including vasodilatation.40 Furthermore, IL-17 induces expression of receptor activator of nuclear factor-jB ligand (RANKL) in osteoblasts;23 RANKL is a membrane-bound receptor of the TNF superfamily that promotes differentiation of osteoclasts. Strikingly, Th17 cells also express elevated levels of RANKL, suggesting that they may be particularly adept at promoting bone turnover.41 Accordingly, IL-17 not only enhances inammation, but stimulates osteoclast differentiation leading to subsequent bone and cartilage damage.42 Although IL-17 alone has the capacity to induce proinammatory factors, its activities are vastly increased when combined with other cytokines, particularly TNF-a (Table 2). This is probably the situation in inamed joints, where multiple inammatory cytokines are over313
Representative target genes Cytokines (IL-6, G-CSF, OSM), chemokines (CXCL1, CXCL2, CXCL5, CCL2, CCL7), transcription factors (C/EBPb, d, IjBf), bone remodelling (RANKL, MMP13), antimicrobial peptides (Lcn2, BDs) IL-6, CXCL8, LIF IL-6, IL-8, RANTES MMP13 Cytokines (IL-19, -20, -24, G-CSF), chemokines (CXCL1), antimicrobial peptides (BDs, S100A7, 8, 9) LL-37 (cathelicidin) Twist1 IL-6, IL-8
expressed.43 Interleukin-17 synergizes with TNF-a to promote induction of nearly all its target genes, and in many cases synergy has also been observed with IFN-c and IL-1b (reviewed in ref. 27). As a consequence, IL-17 alone or together with other inammatory cytokines in the inamed joint mediates adverse events in RA.
which appears to promote spontaneous generation of autoreactive GC and hence elevated autoantibody production.45 Another animal model with lupus-like features is the Ets-1)/) mouse, characterized by increased high-titre autoantibodies that deposit in kidney.48 In these mice, more naive T cells differentiate into a Th17 phenotype.49 Ets-1 is transcription factor expressed in various immune cells and it skews towards a Th17 phenotype by interfering with the ability of IL-2 to inhibit Th17 differentiation. The Ets1)/) mice have low IL-2 production, but T cells from these mice also appear to be refractory to the inhibitory effects of IL-2 on Th17 development. Interestingly RORct expression and IL-2-induced signal transducer and activator of transcription 5 (STAT5) were comparable with wild-type and Ets-1)/) T cells, and so the specic pathway by which Ets-1 exerts its effects is down-stream or independent of STAT5. Interestingly, IL-17 has also been recently shown to synergize with another TNF superfamily member, B-cell activating factor (BAFF), to protect B cells from apoptosis, thereby increasing the number of autoantibodyproducing cells.46 Increased BAFF expression is found in 2225% of SLE serum samples, and BAFF transgenic mice have a lupus-like phenotype.50 Both IL-17R and BAFFR use a common adaptor molecule Act1; however, Act1 is a positive regulator of IL-17R signalling, whereas it is a negative regulator of BAFFR down-stream pathways.51 Following IL-17 and BAFF stimulation of B cells, Act1 is preferentially recruited to IL-17RA, providing an intriguing mechanism for synergistic signalling between these two systems. Subsequently, IL-17 and BAFF together enhance expression of the transcription factor Twist1, which initiates a cascade of gene expression leading to induced expression of the anti-apoptotic genes Twist-2 and B-1, ultimately promoting B-cell differentiation to autoantibody-producing plasma cells46 (Tables 1 and 2). Hence, the impact of IL-17 on B cells may explain its role in contributing to SLE pathogenesis.
express IL-17RA, but start to do so upon differentiation to Th1. Interleukin-17 suppresses IFN-c secretion by repressing T-bet, the master regulator of Th1 cells, which in turn leads to suppression of IFN-c, IL-12Rb1 and osteopontin expression (Table 1). Therefore, in this transfer model, IL17 protects against IBD by limiting Th1 cell activity.65
Psoriasis
Psoriasis is a chronic inammatory skin disorder characterized by dermal hyperplasia. The key histological features of psoriatic skin are epidermal keratinocyte hyperproliferation, vascular proliferation and inltration of DCs, macrophages, neutrophils and T cells.66 The critical roles of IL-23/IL-17 were highlighted in a GWAS study that linked IL-23R polymorphisms to psoriasis, similar to IBD.67 Based on this nding, a model of psoriasis was developed using intradermal injection of IL-23. In this model, anti-IL-17 treatment decreased granulocyte colonystimulating factor (G-CSF) and MMP-13, although it had no effect on erythema, induration and parakeratosis.68 Several IL-10-family cytokines have been found in psoriatic skin, including IL-19, IL-20, IL-22 and IL-24. However, IL-19)/) and IL-24)/) mice still developed skin thickening following IL-23 injection, although mice lacking the common receptor IL-20R2 were resistant.68 In contrast, Zheng et al.69 observed no elevation of IL-19, IL-20 or IL-24 in the IL-23 injection model, whereas IL-22)/) mice exhibited signicant reduction in skin thickness. A role for IL22 is consistent with ndings in human microarray studies, which identify IL-22 as well as many typical Th17 genes.7072 As a consequence, IL-22 from Th17 cells and an IL-20R2-using cytokine have roles in developing psoriasis-like symptoms caused by ectopic IL-23. A mechanism of IL-17 activity in psoriasis likely stems from its co-operative gene regulation IL-22 and other stimuli. Together with IL-17, IL-22 synergistically increases expression of skin antimicrobial peptides, including b-defensin-2 (BD-2), S100A7 (psoriasin) and S100A8/9 (calprotectin).73 Supporting this, S100A79 are elevated in psoriasis, correlating with disease onset.74 Interestingly, psoriasis patients are more resistant to skin infections than people without psoriasis, perhaps as the result of elevated antimicrobial peptide production. Another antimicrobial peptide, cathelicidin (LL37), is synergistically increased by treatment with IL-17 in combination with 1,25-dihydroxyvitamin D3.75 LL37-bound self-DNA fragments trigger TLR9 in DC, which induces a potent adaptive immune response, possibly one of the mechanisms by which self-tolerance is broken.76
Infection
In contrast to its adverse effects in autoimmunity, IL-17 plays a vital role in protecting the host from infection.13 This is particularly evident at mucosal sites such as lung, gut and the oral cavity. Interleukin-17-producing cells are enriched at mucosal surfaces, and Th17 cells express the CCR6 receptor that targets them to mucosal areas.84,85 Pro-inammatory cytokines such as, IL-6, IL-1b and TNF-a, which mediate defensive responses are induced by IL-17. In particular, IL-6, acts in a positive feedback loop to further amplify Th17 differentiation and activate acute-phase responses and complement.28 Interleukin-17 modulates neutrophils via cytokines that promote polymorphonuclear cell expansion and survival (G-CSF, granulocytemacrophage CSF)86 as well as neutrophil chemoattractants (CXCL1, CXCL2 and CXCL5). Additionally, CXCL9, CXCL10 and CCL20 are target genes of IL-17, which have chemotactic activity for lymphocytes, DC and other immune cells, targeting them to mucosal
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Fungal infections
In the setting of fungal infections, IL-17 plays both protective and destructive roles depending on route of infection and perhaps the morphological form of the organism. The primary organism where this has been examined is Candida albicans, a common commensal that colonizes human mucosal surfaces. Candida albicans causes both systemic and mucosal infections, but the immune responses at these sites are quite different.107 The most severe form of candidiasis is a disseminated disease associated with hospital settings, which is modelled by intravenous injection of C. albicans into mice. The IL-17RA)/) mice are highly susceptible to this form of disease. Although detailed analysis of relevant target genes was not performed, neutrophil recruitment was severely defective.108 Consistent with this, two reports using mouse models of oropharyngeal candidiasis (thrush) showed that IL-17 and IL-23 play a strongly protective role against mucosal C. albicans.109,110 Microarray analysis showed that immunocompetent wild-type mice infected with C. albicans up-regulate numerous classic IL-17 target genes in the oral mucosa, including BD3, S100A8/9, MMP-8, G-CSF, CCL20, IL-6, CXCL1 and CXCL5.109 BD3 and S100A8/9, in particular, were strongly impaired in IL-17RA)/) tongue tissue. In contrast, few IFN-c signa317
Viral infections
Viral host defence depends heavily on Type I IFNs that modulate viral replication, and so IL-17 is considered to be relatively less important. However, there is emerging evidence that IL-17 may participate in viral immune responses, which can be benecial or detrimental to the host. It is also intriguing that a homologue of IL-17 is encoded in a Herpesvirus saimiri, a T-cell tropic c herpesvirus.114 The signicance of this is unknown, although viral IL-17 promotes positive signalling through IL-17RA.114 Nonetheless, this homologue presumably benets the virus in some aspect of pathogenesis, by some as yet unknown mechanism. In poxvirus infections, genetically engineered recombinant vaccinia viruses (VV) encoding IL-12, IL-23 or IL-17 were created to test the roles of these cytokines in viral host defence. Surprisingly, VV-IL-23 and VV-IL-17, but not VVIL-12, caused reduced virulence in mice.115 Although the mechanism was not well dened, protection was not mediated by enhanced cytotoxic lymphocyte activity. Surprisingly, IL-23-induced viral resistance was also not primarily the result of IL-17, as IL-17)/) mice infected with VV-IL-23 were not signicantly compromised.115 Interleukin-22 and other Th17-hallmark cytokines were not evaluated in this study. In a contrasting report, an IL-17-expressing VV was found to be more virulent than its parental virus in mice, associated with altered immunoglobulin G isotype generation.116 The distinctions between these models is unclear, but may reect the ne line between host defence and immunopathology mediated by IL-17. Interleukin-17 signalling may be counterproductive in certain viral settings, by contributing to the cytokine
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expanding diversity of effector T cell lineages. Annu Rev Immunol 2007; 25:82152. 10 McGeachy MJ, Cua DJ. Th17 cell differentiation: the long and winding road. Immunity 2008; 28:44553. 11 Bluestone JA, Mackay CR, Oshea JJ, Stockinger B. The functional plasticity of T cell subsets. Nat Rev Immunol 2009; 9:8116. 12 Dong C. TH17 cells in development: an updated view of their molecular identity and genetic programming. Nat Rev Immunol 2008; 8:33748. 13 OQuinn D, Palmer M, Lee Y, Weaver C. Emergence of the Th17 pathway and its role in host defense. Adv Immunol 2008; 99:11563. 14 Yao Z, Spriggs M, Derry J et al. Molecular characterization of the human interleukin (IL)-17 receptor. Cytokine 1997; 9:794800. 15 Yu J, Gaffen SL. Interleukin-17: a novel inammatory cytokine that bridges innate and adaptive immunity. Front Biosci 2008; 13:1707. 16 Firestein GS. Evolving concepts of rheumatoid arthritis. Nature 2003; 423:35661. 17 Brand S. Crohns disease: Th1, Th17 or both? The change of a paradigm: new immunological and genetic insights implicate Th17 cells in the pathogenesis of Crohns disease Gut 2009; 58:115267. 18 Albanesi C, Cavani A, Girolomoni G. IL-17 is produced by nickel-specic T lymphocytes and regulates ICAM-1 expression and chemokine production in human keratinocytes: synergistic or antagonistic effects with IFN-c and TNF-a. J Immunol 1999; 162:494502. 19 Aarvak T, Chabaud M, Miossec P, Natvig JB. IL-17 is produced by some proinammatory Th1/Th0 cells but not by Th2 cells. J Immunol 1999; 162:124651. 20 Gaffen SL. The role of interleukin-17 in the pathogenesis of rheumatoid arthritis. Curr Rheumatol Rep 2009; 11:36570. 21 Brennan FM, McInnes IB. Evidence that cytokines play a role in rheumatoid arthritis. J Clin Invest 2008; 118:353745. 22 Lubberts E. IL-17/Th17 targeting: on the road to prevent chronic destructive arthritis? Cytokine 2008; 41:8491. 23 Kotake S, Udagawa N, Takahashi N et al. IL-17 in synovial uids from patients with rheumatoid arthritis is a potent stimulator of osteoclastogenesis. J Clin Invest 1999; 103:134552. 24 Lubberts E, Koenders MI, Oppers-Walgreen B, van den Bersselaar L, Coenen-de Roo CJ, Joosten LA, van den Berg WB. Treatment with a neutralizing anti-murine interleukin-17 antibody after the onset of collagen-induced arthritis reduces joint inammation, cartilage destruction, and bone erosion. Arthritis Rheum 2004; 50:650 9. 25 Nakae S, Nambu A, Sudo K, Iwakura Y. Suppression of immune induction of collagen-induced arthritis in IL-17-decient mice. J Immunol 2003; 171:61737. 26 Lubberts E, Joosten LA, van de Loo FA, Schwarzenberger P, Kolls J, van den Berg WB. Overexpression of IL-17 in the knee joint of collagen type II immunized mice promotes collagen arthritis and aggravates joint destruction. Inamm Res 2002; 51:1024. 27 Shen F, Gaffen S. Structurefunction relationships in the IL-17 receptor: implications for signal transduction and therapy. Cytokine 2008; 41:92104. 28 Ogura H, Murakami M, Okuyama Y et al. Interleukin-17 promotes autoimmunity by triggering a positive-feedback loop via interleukin-6 induction. Immunity 2008; 29:62836. 29 Shen F, Ruddy M, Plamondon P, Gaffen S. Cytokines link osteoblasts and inammation: microarray analysis of interleukin-17- and TNF-alpha-induced genes in bone cells. J Leukoc Biol 2005; 77:38899. 30 Takemura S, Braun A, Crowson C, Kurtin PJ, Coeld RH, OFallon WM, Goronzy JJ, Weyand CM. Lymphoid neogenesis in rheumatoid synovitis. J Immunol 2001; 167:107280. 31 Timmer TCG, Baltus B, Vondenhoff M, Huizinga TWJ, Tak PP, Verweij CL, Mebius RE, van der Pouw Kraan TCTM. Inammation and ectopic lymphoid structures in rheumatoid arthritis synovial tissues dissected by genomics technology: identication
Acknowledgement
S.L.G. is supported by the National Institutes of Health (AR054389), the Alliance for Lupus Research and Amgen.
Disclosures
SLG has received a research grant and honoraria from Amgen.
References
1 2 Aggarwal S, Gurney AL. IL-17: a prototype member of an emerging family. J Leukoc Biol 2002; 71:18. Gaffen SL. Structure and signalling in the IL-17 receptor family. Nat Rev Immunol 2009; 9:55667.
319
320
113 Zelante T, De Luca A, Bonifazi P et al. IL-23 and the Th17 pathway promote inammation and impair antifungal immune resistance. Eur J Immunol 2007; 37:2695706. 114 Yao Z, Fanslow WC, Seldin MF, Rousseau A-M, Painter SL, Comeau MR, Cohen JI, Spriggs MK. Herpesvirus Saimiri encodes a new cytokine, IL-17, which binds to a novel cytokine receptor. Immunity 1995; 3:81121. 115 Kohyama S, Ohno S, Isoda A et al. IL-23 enhances host defense against vaccinia virus infection via a mechanism partly involving IL-17. J Immunol 2007; 179:391725. 116 Patera AC, Pesnicak L, Bertin J, Cohen JI. Interleukin 17 modulates the immune response to vaccinia virus infection. Virology 2002; 299:5663. 117 Crowe CR, Chen K, Pociask DA et al. Critical role of IL-17RA in immunopathology of inuenza infection. J Immunol 2009; 183:530110. 118 Hou W, Kang HS, Kim BS. Th17 cells enhance viral persistence and inhibit T cell cytotoxicity in a model of chronic virus infection. J Exp Med 2009; 206:31328. 119 Toy D, Kugler D, Wolfson M, Vanden Bos T, Gurgel J, Derry J, Tocker J, Peschon J. Cutting edge: interleukin 17 signals through a heteromeric receptor complex. J Immunol 2006; 177:369. 120 Novatchkova M, Leibbrandt A, Werzowa J, Neubuser A, Eisenhaber F. The STIRdomain superfamily in signal transduction, development and immunity. Trends Biochem Sci 2003; 28:2269. 121 Qian Y, Liu C, Hartupee J et al. The adaptor Act1 is required for interleukin 17dependent signaling associated with autoimmune and inammatory disease. Nat Immunol 2007; 8:24756. 122 Chang S, Park H, Dong C. Act1 adaptor protein is an immediate and essential signaling component of interleukin-17 receptor. J Biol Chem 2006; 281:356037. 123 Liu C, Qian W, Qian Y et al. Act1, a U-box E3 ubiquitin ligase for IL-17 signaling. Sci Signal 2009; 2:ra63. 124 Ruddy M, Wong G, Liu X, Yamamoto H, Kasayama S, Kirkwood K, Gaffen S. Functional cooperation between interleukin-17 and tumor necrosis factor-alpha is mediated by CCAAT/enhancer-binding protein family members. J Biol Chem 2004; 279:255967. 125 Tohyama M, Hanakawa Y, Shirakata Y et al. IL-17 and IL-22 mediate IL-20 subfamily cytokine production in cultured keratinocytes via increased IL-22 receptor expression. Eur J Immunol 2009; 39:277988. 126 Woltman AM, de Haij S, Boonstra JG, Gobin SJ, Daha MR, van Kooten C. Interleukin-17 and CD40-ligand synergistically enhance cytokine and chemokine production by renal epithelial cells. J Am Soc Nephrol 2000; 11:204455. 127 Hartupee J, Liu C, Novotny M, Li X, Hamilton T. IL-17 enhances chemokine gene expression through mRNA stabilization. J Immunol 2007; 179:413541. 128 Zrioual S, Ecochard R, Tournadre A, Lenief V, Cazalis M-A, Miossec P. Genome-wide comparison between IL-17A- and IL-17F-induced effects in human rheumatoid arthritis synoviocytes. J Immunol 2009; 182:311220.
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