慢性乙型肝炎阿德福韦酯治疗应答不佳者外周血单个核细胞HBV-P区基因突变分析  

HBV-P mutations in PBMCs of patients with chronic hepatitis B and a suboptimal response to adefovir treatment

作　　者：高淑春[1] 王晓花[1] 崔蕾[1] 张立新[1] 徐皖苏[1] 安勇[1] 陈士俊[1] 杜以真[1] 

机构地区：[1]山东大学附属传染病医院,山东济南250021

出　　处：《中国病原生物学杂志》2014年第1期5-11,67,共8页Journal of Pathogen Biology

基　　金：济南市2012年科技发展计划(201221051)

摘　　要：目的研究阿德福韦酯（ADV）治疗基因C型慢性乙型肝炎（cHB）患者外周血单个核细胞（PBMcs）乙肝病毒聚合酶区（HBV—P区）序列的基因突变。方法14例基因c型CHB患者分为初治组（ADV初始治疗）7例，经治组（拉米夫定100mg口服每日1次，治疗24～56个月出现YMDD变异后换用ADV治疗）7例，ADV用药时间2～59个月，剂量10mg口服每日1次，血清HBVDNA≥1000copies／ml（应答不佳）者应用直接测序法检测血清及PBMCs内的HBV-P区基因突变，比较两者的异同。结果所有患者的血清和PBMCs内HBV均成功测序，大部分患者血清和PBMCs内的HBV序列一致。1例经治者和3例初治者感染的HBV为野生株，分别有11691＋＋＋V＋、Q215H＋＋＋Q＋、P237T＋＋＋突变。另10例（71．4％）检测到HBV有1个或多个耐药位点变异，主要变异位点为A181V／T／I、N236T。有1例血清HBV基因突变（A181T＋＋＋、N236T＋＋＋）与PBMCs内的HBV基因突变（A181T＋＋＋、N236T＋＋＋、11691＋＋＋V＋）不完全一致。结论ADV治疗cHB应答不佳者大部分血清与PBMCs内的HBV-P区基因突变一致，耐药模式不同，可有一个或多个位点突变，主要变异位点为A181V／T／I、N236T。Objective The aim of this study was to determine mutations of the polymerase gene of hepatitis B virus （HBV-P） in peripheral blood mononuclear cells （PBMCs） of patients with chronic hepatitis B （CHB） who were treated with adefovir dipivoxil （ADV）. Methods Fourteen patients with CHB due to hepatitis B virus （HBV） genotype C had a mean age of 45.64＋9.46 years and were divided into two groups. Nucleos（t）ide analogue （NA）-nalve patients （group A） included 7 patients who received ADV therapy for the first time. Previously treated patients （group B） included the remaining 7 patients who had HBV with YMDD mutations after treatment with [amivudine （100 mg daily） for 24 to 49 months and who had been switched to ADV （10 mg daily） therapy. The duration of ADV treatment ranged from 2 to 59 months. Mutations in HBV-P in sera and PBMCs were detected by direct sequencing in patients with HBV DNA≥1 000 copies/ml. The similarities and differences in those mutations were compared. Results Most samples were sequenced successfully and most HBV gene sequences in sera were in accordance with those in PBMCs. The HBV of 1 previously treated patients and 3 NA-naIve patients were wild-type strains （with I169I＋＋＋V＋, Q215H＋＋＋Q＋, and P237T＋ ＋＋ mutations）. One or more mutations, which were mainly A181 V/T/I and N236T, were noted in samples from the other ten patients. One patient did have serum HBV mutations （A181T＋ ＋＋ and N236T＋＋＋） that were not entirely consistent with those in the patienfs PBMCs （A181T＋＋＋, N236T＋＋＋, and I169I＋＋＋V＋）. Conclusion Mu tations in the HBV-P of HBV in sera were consistent with those in PBMCs from patients with CHB and a suboptimal re sponse to ADV therapy. However, the modes of resistance to ADV were similar in sera and PBMCs. Genotypic resist- ance consisted of one or more amino acid substitutions, most of which were A181 V/T/I and N236T.

关 键 词：慢性乙型肝炎 外周血单个核细胞 基因突变 阿德福韦酯 直接测序 

分 类 号：R512.32[医药卫生—内科学]