【摘要】 目的:研究胶原诱导的关节炎(CIA)大鼠关节中局部浸润的T细胞TCRVβ克隆型。方法:用II型胶原(CII)诱导Lewis大鼠发生关节炎后,经流式细胞术分析大鼠外周血CD4+和CD8+细胞的变化。提取CIA大鼠关节组织的总RNA,通过RT-PCR/SSCP方法分析大鼠两后肢足关节中TCRVβ克隆型的一致率。将CIA大鼠脾淋巴细胞转移至同系正常大鼠,观察发病情况。结果:从注射CII的第30天起,大鼠后爪出现红、肿等症状。流式细胞术分析显示,与正常大鼠相比较,CIA大鼠外周血CD8+T细胞的降低有显著性意义(P<0.01),CD4+T/CD8+T的比值大于2(P<0.01),提示CIA大鼠体内存在T淋巴细胞亚群的比例失调。CIA大鼠两后肢足关节中聚集的TCRVβ克隆型的一致率,以TCRVβ5.2和8.2最高,分别为78.89%和72.23%。脾淋巴细胞转移试验显示,受者发病的大鼠关节中,出现与供者CIA大鼠一致的TCRVβ克隆型条带,表明关节局部聚集的TCRVβ克隆型与关节炎(RA)的发病有关。结论:CIA大鼠关节局部存在以TCRVβ5.2和8.2为主的致病性的T细胞TCRVβ克隆型,可将其作为治疗CIA的靶物质,进行实验动物RA治疗的研究。更多还原

【Abstract】 AIM: To investigate the TCR Vβ clonetypes of T lymphocytes infiltrating in the synovium of rats with collagen-induced arthritis (CIA). METHODS: Type II collagen was used to induce arthritis in Lewis rats. CD4~+ and CD8~+ T lymphocytes in peripheral blood of the rats were analyzed by flow cytometry (FCM). Total RNA was extracted from rat joint tissues and TCR Vβ clonotypes were identified by RT-PCR and single-strand conformation polymorphism (SSCP). Splenic lymphocytes from CIA rats were transferred into normal Lewis rats. TCR Vβ clonotypes of T lymphocytes infiltrating in the joints of the recipient rats were also analyzed by RT-PCR/SSCP. RESULTS: FCM analysis of the T lymphocytes indicated that CD8~+ T lymphocytes decreased in CIA rats and the CD4~+/CD8~+ T lymphocytes ratio was >2, which suggested that there was a disproportion of the T lymphocyte subsets in CIA rats. RT-PCR/SSCP analysis showed that TCR Vβ5.2 and TCR Vβ8.2 clonotypes predominated in the T lymphocytes infiltrating in the joints of CIA rats. T lymphocytes infiltrating in the joints of the recipient rats displayed the same TCR Vβ clonotypes as those in the donor rats. CONCLUSION: CIA rats may have a limited number of dominant TCR Vβ clonotypes of T lymphocytes accumulating in the synovium, of which TCR Vβ 5.2 and 8.2 predominate.更多还原