中国13个省份结核分枝杆菌5种特异性抗原人T细胞表位多态性分析

摘要

Objective To investigate the polymorphisms of the coding gene and the human T cell epitopes of antigen GlnA1,Mpt70,LppX,GroES and LpqH on Mycobacterium tuberculosis complex (MTBC) strains in thirteen provinces of China.Methods A total of 173 clinical MTBC isolates from thirteen provinces were selected to test the gene sequences of the five antigens,using PCR and DNA sequencing methods.Sequences were compared and sliced by BioEdit,and the variations of the human and nonhuman T cell epitopes were analyzed.The rates on synonymous mutation (dS),non-synonymous mutation (dN) and dN/dS values were calculated by Mega 6.0 software.Results Among the 173 strains,there were two non-synonymous mutations in the non-epitope region of glnA1,one non-synonymous mutations in epitope domain of mpt70,one non-synonymous mutation and one synonymous mutation in the epitope domain of lpqH;while groES showed no mutation,lppX had five non-synonymous mutations and one synonymous mutation in the epitope domain.Nine strains presented higher polymorphism at the same gene locus of position 152 in lppX.And seven of the fifteen epitopes contained in lppX were altered and the dN/dS value of this gene was 0.19.Conclusions Data from the human T cell epitope domains of MTBC antigens Mpt70,LppX and LpqH contained epitope diversity,indicated that these antigens may have involved in diversifying the selection to evade the host immunity.GlnA 1 had the polymorphism in epitope domain,which might have little influence on the immuno-response.While GroES seemed relatively conservative,it could play an important role on identification,diagnosis and the development of potential Mycobacterium tuberculosis vaccine.%目的 分析我国结核分枝杆菌复合群(MTBC)临床分离株中GlnA1、Mpt70、LppX、GroES和LpqH 5种抗原编码基因多态性及其人T细胞表位的多态性.方法 选取13个省份临床分离的173株MTBC,采用PCR扩增5种抗原基因,并运用BioEdit软件进行序列比对,分析其人T细胞表位与非表位的变异情况.利用Mega 6.0软件分别计算同义突变率(dS)和非同义突变率(dN)及其比值.结果 173株菌的基因glnA1非表位区出现2个非同义突变位点;基因mpt70表位区出现1个非同义突变位点;基因lpqH表位区表现为1个非同义突变位点和1个同义突变位点;groES在整个基因中未发现任何突变;基因lppX表位区表现为5个非同义突变和1个同义突变位点,其中152位的氨基酸相同位点有9株菌发生了同义突变,表现较高的多态性.同时基因lppX的15个T细胞抗原表位中有7个表位发生了改变,其dN/dS值为0.19.结论 结核分枝杆菌抗原Mpt70、LppX和LpqH的人T细胞表位区具有多态性,反映了此抗原可能参与逃避宿主免疫的分化选择.GlnA1的非表位区的多态性,对该菌株的免疫反应影响较小.GroES序列相对保守,不具有明显的多态性,可能对结核分枝杆菌的鉴定、诊断及新型疫苗的研制具有重要作用.