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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。
Objective To investigate the relationship between type Ⅱ collagen (C Ⅱ) - specific T cell proliferation and C Ⅱ antibody production in patients with rheumatoid arthritis (RA) and human HLA - DR_4 subtype. Methods The CⅡ263 -272 decapeptide was incubated with peripheral blood mononuclear cells (PBMCs) of 62 patients with RA for 5 days. The proliferation of T cells was measured by MTT assay. Serum CⅡ antibody levels were measured by enzyme linked immunosorbent assay (ELISA). Peripheral blood DNA was extracted from 40 patients, and the HLA DR typing was determined by PCR amplification of 33 HLA-DR_4 allelic subtypes. Results In 62 RA patients, 38 (61.3%) T cells in patients after adding C Ⅱ 263-272 significant proliferation, C Ⅱ 263 -272 antibody positive rate of 53.2%. The positive rate of CⅡ263-272 antibody was 66.7% in patients with positive cell proliferation reaction, 34.6% in patients with negative proliferative response, P <0.05. The positive rate of T cell proliferation was 69.7% in CⅡ263-272 antibody positive patients and 42.1% in negative patients, P <0.05. All patients with T cell proliferation stimulation index (SI) and C Ⅱ 263-272 antibody concentration was significantly correlated (r = 0 68, P <0.01). Of the 40 patients, HLA-DR_4 alleles were found in 16 cases of HLA typing. HLA-DR4 was not found to be associated with CII-specific T cell proliferation and CII antibody positive. Conclusion The formation of C Ⅱ antibody may be related to the activation of B cells induced by C Ⅱ specific T cells.