基于免疫分选的T淋巴细胞亚群嵌合性定量分析

Quantitative chimerism analysis of T lymphocyte subsets based on immune sorting

目的:建立基于免疫分选的T淋巴细胞亚群嵌合性定量分析的方法。方法:以淋巴细胞不同比例混合制备14组人工嵌合体血样,以免疫磁珠阳性选择法获取CD4+和CD8+T细胞亚群,分选后的细胞再进行16位点的短串重复序列(STR)多态性分析。结果:分选后T细胞亚群提取的DNA量能满足STR检测要求,当嵌合体中受体比例≥10%时,16个STR位点均能检出受体和供体特异性等位基因,按照STR峰面积计算的受体比例与理论值相符;当受体比例≤1%时,仅有部分位点能检出受体特异性等位基因,实测嵌合率与理论值存在差异。结论:建立了基于免疫分选的T细胞亚群嵌合性定量分析方法,可用于造血干细胞移植术后嵌合体监测。

Objective：To establish a quantitative method for chimerism analysis of T lymphocyte subsets based on immune sorting.Methods： 14 artificial chimera groups were mixed according to different lymphocytes proportion.The CD4＋ and CD8＋ T cell subsets were sorted by positive selection of immunomagnetic beads,and then were analyzed by STR polymorphisms technique.Results： The DNAs extracted from sorted T cells were fit for STR detection.All STR alleles specific for recipient or donor can be detected and the quantitative results were consistent with theoretic values in over 10% recipient chimeras.But only partial recipient alleles can be detected and the quantitative results were different from theoretic values in less then 1% recipient chimeras.Conclusion： A quantitative chimerism analysis of T cell subsets based on immune sorting was established.The method can be used to monitoring of hematopoietic chimerism following hematopoietic stem cell transplantation.