系统性红斑狼疮自体外周血干细胞移植T细胞克隆基因谱型变化  

The variation of T-cell clonal repertoire in patients with systemic lupus erythematosus (SLE) following autologous peripheral blood hematopoietic stem cell transplantation

作　　者：符粤文[1] 朱平[2] 赵晓武[1] 李惠芳[3] 曾中州[1] 张蕊[4] 王奕嘉[3] 张英[3] 刘继华[4] 

机构地区：[1]郑州市第三人民医院,450001 [2]北京大学第一医院血液病实验室,100034 [3]北京大学第一医院 [4]北京儿童医院

出　　处：《中华医学杂志》2003年第19期1648-1652,共5页National Medical Journal of China

基　　金：国家自然科学基金 (3 9970 13 1);北京市自然科学基金 (7992 0 3 9);北京大学人类疾病中心基金 ;北京大学生物医学跨学科基金;北京大学肿瘤中心科研基金

摘　　要：目的　通过系统性红斑狼疮 (SLE)自体外周血干细胞移植 (APBSCT)前后T细胞受体 β(TCRβ)CDR3基因表达谱型的变化 ,探讨SLE发病机理以及患者干细胞移植后T淋巴细胞克隆的免疫重建特征。方法　应用RT PCR扩增APBSCT的SLE患者外周血的TCRβ可变区 (V) 2 5个家族的基因序列 ,在长泳道测序胶上电泳 ,形成TCRβCDR3基因表达谱型图。通过谱型图上电泳条带分析SLE移植前T细胞克隆增生的特征以及移植后T细胞克隆群的变化 ,做增生T细胞克隆的TCRβ基因序列分析 ,了解TCRβ基因与自身免疫病的相关性。 结果　正常TCRβV2 5个家族的基因序列电泳后分别出现呈高斯分布的 10余条电泳条带 ,组成CDR3基因表达谱型图。 8例SLE患者谱型图均出现异常 ,4例TCRβ在 βV13.1基因家族T细胞出现寡克隆增生 ,3例在 βV8,βV9,βV15家族中出现寡克隆性增生。APBSCT后T细胞克隆分布趋于正常 ,CDR3恢复多态性 ,部分恢复到正常TCRβ基因分布。测序结果显示谱型图上单一条带过度浓集往往是单克隆或者寡克隆T细胞增殖 ;发现SLE一个异常T细胞克隆的TCRβV8CDR3氨基酸序列与已报道强直性脊椎炎疾病相关克隆完全一致。 结论　SLE患者出现T细胞寡克隆增生 ,增生的T细胞克隆可能与自身免疫病发病有关。APBSCT后免疫重建 ,抑制了致病的T细胞?Objective Analyze the variation of T cell receptorβ(TCRβ) CDR3 gene expressing repertoire in systemic lupus erythematosus (SLE) patients before and after autologous peripheral blood hematopoietic stem cell transplantation (APBSCT), to search the pathogenesis of SLE and characteristics of T cell reconstitute immune after APBSCT. Methods Reverse transcriptase - polymerase chain reaction (RT-PCR) amplified 25 subfamily genes of TCRβ prom peripheral blood lymphocytes (PBLs) of SLE patient with APBSCT and run denaturation polyacrylamide sequencing gel electrophoresis, set up a map of TCRβCDR3 gene expressing repertoire. The bands of TCRβ gene repertoire in the electrophoresis gel can be used to analyze the variety of T cell clones which expanded and disappeared in cases of SLE. Cut the bands and sequencing. Compare sequences of TCRβgenes in normal and abnormal T cell clones to understand the relationship between TCRβ gene and autoimmune diseases. Results In the normal control group, TCRβ gene repertoires show more than 10 bands in each of 25 TCRβV families, characterized as Gaussian distribution. TCRβ gene repertoire in the 8 patients with SLE were abnormally. Four patients expressed TCRβV13.1 gene preferentially. Three patients had TCRβV8, TCRβV9 and TCRβV18 genes over expressed. These over expressive genes represented the oligoclonal expansion of T cell populations. After APBSCT the TCRβ gene repertoires regained CDR3 polymorphism in 5 patients, some of them recovered to normal completely. The sequencing analyse show dense and strong band in map of TCRβ gene repertoires were T cell monoclonal or oligoclonal expansion frequently. In a case of SLE an amino acid sequence of TCRβV8 CDR3 was same to a sequence of disease correlative T cell clone with ankylosing spondylitis that had been reported in a literature before. Conclusion SLE patients have oligoclonal expansion of T cell that related with the pathogenesis of autoimmune disease. After APBSCT, T cell clones recover to normal distribution. T

关 键 词：系统性红斑狼疮 自体外周血干细胞移植 T细胞 克隆 基因表达谱 免疫重建 RT-PCR 

分 类 号：R593.241[医药卫生—内科学]