类风湿关节炎患者外周血T细胞抑制性多肽的研究

目的探讨HLADRB1结合性流感病毒血凝素(HA)308317变构肽对类风湿关节炎(RA)患者外周血T细胞激活的影响。方法选取27例HLADRB1阳性的RA患者,密度梯度离心法分离外周血单个核细胞。采用固相法合成HA308317变构肽,并检测RA患者外周血T细胞对这些多肽的反应性;及多肽对白细胞介素(IL)2、γ干扰素分泌及CD69表达的影响。同时进一步观察HA308317变构肽对CII263272或HA308317多肽介导的T细胞激活的抑制作用。结果变构肽刺激T细胞增殖的阳性率分别为7.4%(APL1)、3.7%(APL2)和3.7%(APL3),明显低于CII263272和HA308317原型肰(62.2%和52.9%),3条变构肽的刺激指数也明显低于原型肽(均P<0.01)。并且,其诱导IL2、γ干扰素分泌的水平及CD69的表达亦下降(P<0.05)。与单独加入CII263272或HA308317原型肽的对照组相比,加入HA308317变构肽的T细胞增殖的刺激指数明显下降(P<0.05)。结论替换HA308317多肽中T细胞受体的接触残基产生了弱或非T细胞刺激肽,它们可能有效地抑制RA的T细胞免疫反应。

The study of inhibitory peptides on T cell activation in rheumatoid arthritis

OBJECTIVE: To study the inhibitory role of altered HA308 - 317 peptides in T cell responses in patients with rheumatoid arthritis (RA). METHODS: Peripheral blood mononuclear cells (PBMC) were obtained from 27 HLA-DRB1 positive RA patients. T cell responses to altered HA308-317 peptides were examined by using (3)H incorporation assay. The level of IL-2 and IFNgamma in the supernatants was identified by an enzyme-linked immunosorbent assay. The CD69 expression on T cell surface was studied by using flow cytometry. RESULTS: Compared to wild type CII263 - 272 or HA308 - 317, altered HA308 - 317 peptides failed to stimulate T cell proliferation (P < 0.05) and production of IL-2 as well as IFNgamma and resulted in lower expression of CD69 in RA patients (P < 0.05). SI values for T cell coincubated with altered HA308 - 317 peptides and CII263 - 272 or wild type HA308 - 317 were significantly lower than coincubated with CII263 - 272 and wild type HA308 - 317 alone (P < 0.05). CONCLUSION: Substitution of TCR-binding residue of HA308 - 317 yielded weak or non-T-cell stimulating peptides, which might be potentially effective in blocking abnormal T cell responses in RA.