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Restrictions of T cell receptor beta chain repertoire in the peripheral blood of patients with systemic lupus erythematosus.

机译:系统性红斑狼疮患者外周血中T细胞受体β链库的限制。

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摘要

OBJECTIVE: To investigate whether oligoclonal T cell populations occur in peripheral blood lymphocytes from patients with systemic lupus erythematosus (SLE). METHODS: RNA was extracted from the lymphocytes isolated from whole peripheral blood of five female patients fulfilling ARA criteria for SLE and two healthy female controls, and synthesised into cDNA. CDR3 length spectratyping was performed using a polymerase chain reaction (PCR) run to saturation followed by a primer extension with a radioactively labelled primer. The resulting samples, one for each of the 23 V beta families, were then run on a polyacrylamide sequencing gel to examine the T cell receptor beta chain repertoire at the level of VDJ length heterogeneity. RESULTS: The two healthy female controls showed faint oligoclonal bands in only two and three V beta families respectively. Three of the patients showed a similar degree of oligoclonality to the controls, while the other two, who had active disease as shown by SLAM scores of 17 and 19 and in one case low C4 and raised C3dg levels, showed marked oligoclonality of their T cell beta chain repertoire affecting more than 17 of the 23 V beta families analysed. CONCLUSIONS: Using the technique of CDR3 length spectratyping, restriction of T cell receptor beta chain usage by peripheral blood T cells in patients with SLE has been demonstrated for the first time.
机译:目的:探讨系统性红斑狼疮(SLE)患者外周血淋巴细胞中是否存在寡克隆T细胞群。方法:从五名符合SLE ARA标准的女性患者和两名健康女性对照的全外周血淋巴细胞中提取RNA,然后合成为cDNA。使用进行至饱和的聚合酶链反应(PCR)进行CDR3长度光谱分型,然后使用放射性标记的引物延伸引物。然后,将所得样品(针对23 Vβ家族中的每个样品)在聚丙烯酰胺测序凝胶上电泳,以检查VDJ长度异质性水平上的T细胞受体β链库。结果:两个健康的女性对照仅分别在两个和三个V beta家族中显示出微弱的寡克隆带。其中三名患者表现出与对照组相似的寡克隆性程度,而其他两名患者的活动性疾病表现为SLAM评分为17和19,其中一种C4低而C3dg水平升高,表现出明显的T细胞寡克隆性beta链库影响了23个V beta系列中超过17个的分析。结论:使用CDR3长度谱分析技术,首次证明了SLE患者外周血T细胞对T细胞受体β链使用的限制。

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