RESUMO
IL-18 is an important cofactor in Th1 immune responses and it has additional roles in inflammation. Recent reports suggest the contribution of IL-18 to immune responses may vary between mouse strains and immune contexts. We investigated the contribution of IL-18 to T-cell activation and joint inflammation in Ag-induced arthritis (AIA) in C57Bl/6 mice. AIA and cutaneous delayed-type hypersensitivity (DTH) reactions were induced in wild-type (WT) and IL-18-/- C57Bl/6 mice, and Ag-specific T-cell proliferation and IFN-gamma and IL-4 production were measured. The humoral immune response was measured as serum antibody to the disease-initiating Ag, methylated BSA (mBSA). Splenocyte production of IL-6 was measured by ELISA. To confirm the dependence of this model on Th1-cell-mediated immunity, IL-12p40-/- mice were similarly studied. WT mice developed synovitis, joint effusion, cartilage destruction and bone damage associated with induction of DTH, and in vitro Ag-specific T-cell proliferation and IFN-gamma production. Unexpectedly, IL-18-/- mice developed AIA and indices of T-cell activation were similar to those of WT mice. In contrast, IL-12p40-/- mice did not develop AIA, DTH or T-cell activation. WT and IL-18-/- mice, but not IL-12p40-/- mice, developed significantly increased serum antibody to mBSA compared with naive controls. WT and IL-18-/- splenocytes produced high levels of IL-6, whereas IL-12p40-/- cells had significantly lower IL-6 production compared with both. In conclusion, IL-18 is redundant both as a Th1 response cofactor and inflammatory cytokine, whereas IL-12p40-/- is a key cytokine, in AIA in C57Bl/6 mice.
Assuntos
Artrite Experimental/imunologia , Interleucina-18/imunologia , Ativação Linfocitária/imunologia , Células Th1/imunologia , Animais , Formação de Anticorpos/genética , Formação de Anticorpos/imunologia , Artrite Experimental/genética , Artrite Experimental/patologia , Autoanticorpos/imunologia , Autoantígenos/imunologia , Citocinas/genética , Citocinas/imunologia , Hipersensibilidade Tardia/genética , Hipersensibilidade Tardia/imunologia , Hipersensibilidade Tardia/patologia , Inflamação/imunologia , Inflamação/patologia , Interleucina-18/genética , Camundongos , Camundongos Knockout , Especificidade da Espécie , Sinovite/genética , Sinovite/imunologia , Sinovite/patologia , Células Th1/patologiaRESUMO
OBJECTIVE: Protease-activated receptor 1 (PAR-1) is the cell surface receptor for thrombin. It is unclear whether thrombin contributes to inflammation other than by effects on coagulation. We investigated the proinflammatory participation of PAR-1 in antigen-induced arthritis (AIA). METHODS: Arthritis was induced by intraarticular injection of methylated bovine serum albumin (mBSA) in preimmunized PAR-1-deficient (PAR-1(-/-)) and wild-type (WT) mice. The disease was assessed after 7 days by histologic examination of knee joints after decalcification and Safranin O/toluidine blue staining. Serum levels of anti-mBSA IgG, interferon-gamma, and interleukin-4 (IL-4) were determined by enzyme-linked immunosorbent assay. T cell proliferation response was determined by measuring the incorporation of (3)H-thymidine. Cytokine messenger RNA (mRNA) expression was detected in synovial tissues and peritoneal cells by real-time polymerase chain reaction. RESULTS: Arthritis severity was significantly reduced in PAR-1(-/-) mice compared with WT mice (P = 0.017). Analysis of individual aspects of joint histology revealed significant reductions in synovial exudates (P < 0.001), cartilage degradation (P < 0.01), and bone damage (P = 0.05) in PAR-1(-/-) mice. Synovial IL-1, IL-6, and matrix metalloproteinase 13 (MMP-13) mRNA was significantly reduced in PAR-1(-/-) mice. The titers of antigen-specific serum anti-mBSA total IgG, IgG1, and IgG2a were significantly reduced, and serum IL-4 was significantly increased in arthritic PAR-1(-/-) mice. In contrast, no difference was observed in antigen-induced T cell proliferation between PAR-1(-/-) and WT mice. In vitro, thrombin-induced (but not lipopolysaccharide-induced) IL-1, IL-6, and MMP-13 mRNA expression was significantly impaired in PAR-1(-/-) mice compared with WT controls. CONCLUSION: These data demonstrate the requirement of PAR-1 for the expression of AIA, the development of an antigen-specific Ig response, thrombin-induced macrophage cytokine and MMP expression, and the inhibitory effect of PAR-1 on serum IL-4. We conclude that PAR-1 plays a significant role in this model of arthritis.
Assuntos
Artrite Experimental/genética , Artrite Experimental/patologia , Articulações/patologia , Receptor PAR-1/deficiência , Receptor PAR-1/genética , Animais , Artrite Experimental/imunologia , Proliferação de Células , Modelos Animais de Doenças , Membro Posterior , Imunoglobulina G/sangue , Interferon gama/sangue , Interleucina-4/sangue , Articulações/imunologia , Macrófagos Peritoneais/citologia , Macrófagos Peritoneais/imunologia , Masculino , Camundongos , Camundongos Knockout , RNA Mensageiro/metabolismo , Receptor PAR-1/imunologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Soroalbumina Bovina/imunologia , Membrana Sinovial/metabolismo , Membrana Sinovial/patologia , Linfócitos T/citologia , Linfócitos T/imunologiaRESUMO
Tissue factor (TF), a transmembrane receptor for plasma factor VII(a), is the main initiator of the coagulation cascade. It has also been implicated in noncoagulant processes, including inflammation. The function of the TF cytoplasmic domain was studied in mice in which 18 of the 20 cytoplasmic amino acids were deleted. This mutation (TF(deltaCT/deltaCT)) is not associated with alterations in blood coagulation. Arthritis was induced by intra-articular injection of methylated bovine serum albumin (mBSA) in mice preimmunized with mBSA. Arthritis severity was significantly reduced in TF(deltaCT/deltaCT) mice compared to wild-type mice, including reductions in synovitis, synovial exudate, cartilage degradation, and bone damage. A marked reduction in synovial interleukin (IL)-1beta and IL-6 mRNA was also observed. Serum anti-mBSA IgG1, but not IgG2a, was increased in mutant mice. Cutaneous delayed-type hypersensitivity and antigen-induced T-cell proliferation were reduced in TF(deltaCT/deltaCT) compared to wild-type mice. A significant down-regulation of lipopolysaccharide-induced IL-1, tumor necrosis factor, IL-6, macrophage migration inhibitory factor, and matrix metalloproteinase-13 mRNA was observed in immunized, but not in naive TF(deltaCT/deltaCT) macrophages ex vivo. These data suggest a significant role for the cytoplasmic domain of TF in the regulation of the immunoinflammatory responses, a murine arthritis model, and macrophage function.