HIV/HBV合并感染者HBV核心区与多聚合酶区特异性细胞毒性T细胞表位变异分析  

Analysis the characteristic mutation of specific cytotoxic T lymphocytes epitopes in HBV core protein and polymerase protein in HIV/HBV co-infected patients

作　　者：邓西子 聂源 兰芸[1] 李锋 高鸣 蔡卫平[1] 李凌华[1] 胡凤玉[1] DENG Xizi;NIE Yuan;LAN Yun;LI Feng;GAO Ming;CAI Weiping;LI Linghua;HU Fengyu(The Institute for Infections Disease,Guangzhou Eighth People's Hospital Affiliated toGuangzhou Medical University,Guangzhou Guangdong 510060,China)

机构地区：[1]广州医科大学附属广州市第八人民医院传染病研究所

出　　处：《转化医学杂志》2019年第5期300-303,共4页Translational Medicine Journal

基　　金：“十三五”国家科技重大专项(2017ZX10202101)

摘　　要：目的比较人免疫缺陷病毒(human immunodeficiency virus,HIV)/乙型肝炎病毒(hepatitis B virus,HBV)合并感染者与HBV单一感染者外周血HBV核心(core,C)蛋白和多聚合酶(polymerase,P)蛋白的特异性细胞毒性T细胞(cytotoxic T lymphocyte,CTL)表位变异的差异,探讨HIV/HBV合并感染者肝病进程加快的可能致病机制。方法收集慢性HBV感染者的基本检查结果和血标本,并依据抗病毒前HIV抗体检测结果,分为HIV/HBV合并感染组和HBV单一感染组。提取HBV DNA,巢式PCR法扩增HBV C基因与P基因并送PCR产物测序,ContigExpress软件进行序列拼接,Vector NTI软件进行序列比对,参照相应基因型标准序列,分析2组患者的HBV CTL表位变异差异。结果成功扩增的HBV C基因与P基因的患者共151例,其中HIV/HBV合并感染者83例,HBV单一感染者68例。HIV/HBV合并感染组较HBV单一感染组CTL表位变异发生率均偏高,其中C18-27、C88-96、C139-148表位变异发生率差异比较有统计学意义(χ^2=7.509,P=0.006;χ^2=3.902,P=0.048;χ^2=4.238,P=0.040)。HIV/HBV合并感染组的C18-27表位主要变异氨基酸的类型增多,出现新的S26N(4.8%)、S26T(4.8%)变异类型,而C88-96、C139-148表位主要变异氨基酸相同(L95I、T147A)。结论HIV/HBV合并感染可增加HBV C蛋白和P蛋白的CTL表位变异,尤其是C18-27、C88-96、C139-148表位变异。C蛋白CTL表位变异增多可能与HIV/HBV合并感染肝病进程加快的致病机制相关。Objective To compare the difference in mutations of specific cytotoxic T lymphocytes(CTL)epitopes in hepatitis B virus(HBV)core(C)protein and polymerase(P)protein between human immunodeficiency virus(HIV)/HBV co-infected patients and HBV mono-infected patients,and to explore mechanism of accelerated liver disease progression in HIV/HBV co-infected patients.Methods The basic test results and blood samples of patients with chronic HBV infection were collected and classified into HIV/HBV co-infected group and HBV mono-infected group acc-ording to the results of HIV antibody before treatment.HBV DNA in serum was extracted.C gene,P gene were amplified by nest-PCR and PCR products were sequenced.ContigExpress software was used for sequence splicing and Vector NTI software was used for sequence alignment.With reference to the standard sequence of the matched genotype HBV,differences in CTL epitopes’mutation between the two groups were analyzed.Results HBV C gene and P gene fragments were successfully amplified from 151 patients,including 83 cases of HIV/HBV co-infected group and 63 cases of HBV mono-infected group.The result of analyzing mutants of C protein and P protein indicated that the incidence of mutation in all epitopes for CTL was higher in the HIV/HBV co-infected group,and C18-27,C88-96,C139-148 epitope were statistically significant higher(χ^2=7.509,P=0.006;χ^2=3.902,P=0.048;χ^2=4.238,P=0.040).In the HIV/HBV co-infected group,the major mutation amino acid types of C18-27 epitope increased,new S26N(4.8%)and S26T(4.8%)variant types appeared,however the major mutation amino acids of C88-96 and C139-148 epitopes were the same(L95I,T147A).Conclusion Co-infection of HIV/HBV increased the CTL epitopes’mutations in the HBV C protein and P protein,especially the epitope mutation of C18-27,C88-96 and C139-148.The increase of CTL epitope’s mutation of C protein may be related to the pathogenic mechanism of accelerated progression of HIV/HBV co-infection.

关 键 词：人类免疫缺陷病毒 肝炎病毒 乙型 细胞毒性T细胞 变异 核心蛋白 多聚酶蛋白 

分 类 号：R512.62[医药卫生—内科学]