摘要
目的评价HBV G1613A和C1653T变异对乙型肝炎患者疾病进展、病毒体外复制力及核心启动子(CP)转录活性的影响。方法共纳入258例研究对象,包括65例急性乙型肝炎(AHB)患者,120例慢性乙型肝炎(CHB)患者和73例慢加急性肝衰竭(ACLF)患者。从患者血清中提取HBV DNA,PCR扩增HBV DNA全长基因组,统计G1613A、C 1 6 5 3 T和G 1 6 1 3 A+C 1 6 5 3 T变异的发生率。构建相应载体进行体外功能实验,观察病毒质粒转染He p G 2细胞后对病毒复制力及其CP转录活性的影响。结果 258例患者中共检出B、C、D三种基因型,其检出率分别是22.2%、76.2%和1.6%。G1613A、C1653T及G1613A+C1653T变异发生率随疾病程度加重依次升高。AHB、CHB和ACLF患者上述3种变异的检出率分别为13.70%、31.80%和45.20%(P<0.01),2.30%、16.30%和27.40%(P<0.01),2.29%、12.07%和23.29%(P<0.05)。与野生株相比,G1613A变异株复制力升高6%,HBs Ag降低15%,HBe Ag表达呈阴性,CP转录活性降低16.2%;C1653T变异株复制力升高10%,HBs Ag升高55%,HBe Ag与野生株接近,CP转录活性升高17.1%;G 1 6 1 3 A+C 1 6 5 3 T变异株复制力升高7%,H B s A g升高6 6%,H B e A g升高2 2 7%,但对CP转录活性没有影响。结论 G1613A、C1653T在CP区的变异可增加HBV复制力,影响CP转录活性和HBV抗原的表达,G1613A+C1653T联合变异可能对这些功能产生协同作用,推测这三种变异与乙肝重症化发生机制相关。
Objective To evaluate the impact of hepatitis B virus（HBV） genome G1613 A and C1653 T mutations on disease progression, viral replication capacity, and transcription activity of HBV core promoter（CP）. Methods A total of 258 patients were enrolled in the present study, including 65 patients with acute hepatitis B（AHB）, 120 with chronic hepatitis B（CHB）, and 73 with acute on chronic liver failure（ACLF）. Serum HBV DNA was extracted from patients, and full-length HBV genome was amplified by PCR. The incidences of G1613 A, C1653 T and G1613A＋C1653T in different groups were compared, and through functional experiments, the impact of mutants and wild-type virus on viral replication capacity and CP transcription activity was assessed. Results Genotype B, C and D were the three detected genotypes in 285 patients, with detection rates of 22.2%, 76.2% and 1.6%, respectively. The incidences of G1613 A, C1653 T and G1613A＋C1653T mutations increased with the disease exacerbation, and they were 13.70%, 31.80% and 45.20% in AHB patients（P〈0.01）, 2.30%, 16.30% and 27.40% in CHB patients（P〈0.01）, and 2.29%, 12.07% and 23.29% in ACLF patients（P〈0.05）. Compare with wild-type strain, the G1613 A mutant strain of HBV increased the viral replication capacity by 6%, reduced HBs Ag level and core promoter activity by 15% and 16.2%, and reduced HBe Ag to undetectable level; the C1653 T mutant strain increased the viral replication capacity, HBs Ag level, and core promoter activity by 10%, 55% and 17.1%, respectively, and the HBe Ag level was comparable to that of wild-type strain; the G1613A＋C1653T mutant strain increased viral replication capacity, HBs Ag level and HBe Ag level by 7%, 66% and 227%, respectively, while it had no influence on core promoter activity. Conclusion The G1613 A and C1653 T mutation in CP region may increase HBV replication capacity and alter CP activity and HBV antigens expression, the doublet mutation of G1613A＋C1653T shows synergic effect on these changes, suggesting these mutations are associated with liver disease progression.
出处
《解放军医学杂志》
CAS
CSCD
北大核心
2016年第3期204-210,共7页
Medical Journal of Chinese People's Liberation Army
基金
国家自然科学基金(81271847)~~
关键词
乙型肝炎病毒
启动区
遗传
遗传变异
肝功能衰竭
hepatitis B virus
promoter regions
genetic
genetic variation
liver failure
