胶原诱导的关节炎大鼠关节中浸润的T细胞TCR Vβ克隆型分析

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TCR Vβ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TCR Vβ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TCR Vβ克隆型的一致率，以TCR Vβ5．2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TCR Vβ克隆型条带，表明关节局部聚集的TCR Vβ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TCR Vβ5．2和8．2为主的致病性的T细胞TCR Vβ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。

Analysis of TCR Vβ clonotypes of T lymphocytes infiltrating in the joints of rats with collagen-induced arthritis

AIM: To investigate the TCR Vbeta 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 Vbeta clonotypes were identified by RT-PCR and single-strand conformation polymorphism (SSCP). Splenic lymphocytes from CIA rats were transferred into normal Lewis rats. TCR Vbeta 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 Vbeta5.2 and TCR Vbeta8.2 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 Vbeta clonotypes as those in the donor rats. CONCLUSION: CIA rats may have a limited number of dominant TCR Vbeta clonotypes of T lymphocytes accumulating in the synovium, of which TCR Vbeta5.2 and 8.2 predominate.