【摘要】 目的分析MRL/lpr和(NZB×NZW)F1两种狼疮鼠成年发病时,广泛浸润于多脏器的T细胞克隆T细胞受体(TCR)Vβ基因编码的氨基酸基序特点,揭示该T细胞克隆的抗原特异性。方法用反转录聚合酶链反应(RT-PCR)分别扩增不同成年发病狼疮鼠个体的肾脏、大脑中的TCRVβ6基因,对扩增产物进行基因克隆和DNA测序分析;单链构象多态性分析法(SSCP)鉴定广泛浸润的T细胞克隆TCRVβ6基因编码的氨基酸基序。结果两种成年发病狼疮鼠的不同个体之间,广泛浸润于多脏器的T细胞克隆TCRVβ6基因的互补决定区3(CDR3)中出现相同的氨基酸基序。结论不同个体成年发病狼疮鼠的多脏器中广泛浸润的T细胞克隆TCRVβ基因的CDR3中出现相同的氨基酸,表明该克隆针对特定的自身抗原。更多还原

【Abstract】 Objective There were some identical T cell clonotypes expanded and accumulated in different organs in aged diseased (NZB×NZW)F1 and MRL/lpr mice. The amino acid motifs in the third complementarity determining region (CDR3) from the T cell receptor (TCR) Vβ chain was analyzed to demonstrate the antigenic specificity of these identical clones. Methods The TCR Vβ6 gene in kidneys and brains from different individuals of aged diseased lupus mice were amplified by reverse transcriptase polymerase chain reaction (RT-PCR) respectively. The amino acid motifs in CDR3 loops of TCR Vβ6 chain were demonstrated by DNA cloning and DNA sequence analysis. The amino acid motifs from the identical T cell clones accumulated in different organs were identified by single-strand conformation polymorphism (SSCP). Results Some conserved amino acid motifs such as isoleucine (I) and aspartic acid (D) were observed in CDR3 loops of TCR Vβ6 from these identical T cell clones in different individuals of aged diseased (NZB×NZW)F1 mice. Likewise, I, glycine (G) and D or glutamic acid (E) were also found in MRL/lpr mice. Conclusion These identical T cell clones may recognize restricted T cell epitopes on autoantigens and are involved in specific immune responses in SLE.更多还原