类风湿关节炎患者Ⅱ型胶原特异性T细胞增殖及抗体生成与HLA-DR4的关系

目的　探讨类风湿关节炎(RA)患者Ⅱ型胶原 (CⅡ)特异性T细胞增殖和CⅡ抗体生成,及与人类白细胞抗原(HLA)- DR_4亚型之间的关系。方法　将CⅡ263 -272十肽与 62例RA患者外周血单个核细胞(PBMC)共孵育 5d,应用四甲基偶氮唑盐法测定T细胞增殖。酶联免疫吸附试验(ELISA)法测定患者血清CⅡ抗体水平。提取 40例患者外周血基因组DNA,以 33种HLA- DR_4等位基因亚型的特异引物PCR扩增后测定患者HLA DR分型情况。结果　在 62例RA患者中, 38例(61 .3% )患者的T细胞在加入CⅡ263- 272后出现明显增殖,CⅡ263 -272抗体阳性率为 53. 2%。细胞增殖反应阳性的患者CⅡ263- 272抗体的阳性率为 66. 7%,增殖反应阴性的患者为 34 6%,两者比较P<0 .05。CⅡ263 -272抗体阳性的患者T细胞增殖反应率为 69 7%,阴性患者为 42 .1%, 两者比较P<0 .05。所有患者T细胞增殖刺激指数 (SI)与CⅡ263- 272抗体浓度呈显著正相关 (r=0 68,P<0. 01)。40例患者HLA分型中,有 16例携带HLA -DR_4等位基因,未发现HLA DR_4与CⅡ特异性T细胞增殖反应、CⅡ抗体阳性有显著相关性。结论　CⅡ抗体的生成可能与CⅡ特异性T细胞介导的B细胞活化有关。

Proliferation of type II collagen specific T cell response and antibody formation in rheumatoid arthritis and their relations to HLA-DR4

OBJECTIVE: To investigate the proliferation of type II collagen specific T cell response and antibody formation in rheumatoid arthritis (RA) and their relations to HLA-DR4 subtype. METHODS: Peripheral blood mononuclear cells and serum were obtained from 62 RA patients, 14 males and 48 females, aged 43 +/- 29. CII263-272 decapeptide and the peripheral blood mononuclear cells (PBMCs) from 62 RA patients were co-incubated for 5 approximately 7 days. MTT method was used to examine the T cell proliferation. The serum antibodies specific to CII 263-272 were examined by ELISA. HLA-DR typing was detected in 40 patients by SSP-PCR. RESULTS: Positive T cell response challenged by CII 263-272 decapeptide was observed in 38 (61.3%) RA patients. The positive rate of CII 263-272 antibody was 66.7% in the T cell response positive group, significantly higher than that of the T cell response negative group (34.6%, P < 0.05). The T cell response rate of the CII 263-272 antibody positive group was 69.7%, significant higher than that of the negative group (42.1%, P < 0.05). In all patients, the stimulation index (SI) of T cell proliferation to CII263-272 peptide was positively correlated with the level of antibody (r = 0.68, P < 0.01). HLA-DR4 allele was detected in 16 out of the 40 patients. Positive T cell response and presence of antibody to CII 263-272 were not associated with HLA-DR4 phenotype. CONCLUSION: The formation of CII antibody may be related to B cell activation mediated by CII specific T cell.