丙型肝炎病毒1b型F蛋白抗原和抗体特异性表达的研究

Specific antigen and antibody expression of hepatitis C virus genotype 1b F protein in chronic hepatitis C patients

目的研究丙型肝炎病毒(hepatitis C virus,HCV)1b型F蛋白抗原在慢性HCV感染者中产生的特异性抗原和抗体的表达并检测其流行率。方法应用人工合成HCV 1b型的双框移动F蛋白多肽,采用ELISA方法,对其在慢性HCV感染者中F蛋白的流行率进行分析;合成并纯化抗HCV 1b型F蛋白的抗体,采用Western blot方法验证抗体的特异性,包被纯化的F蛋白的特异性抗体,检测慢性HCV患者血清中F蛋白抗原的表达及其流行率。结果 HCV 1b型F蛋白的抗体在32例HCV 1b型慢性感染者中有19例可以检测到抗体的表达,其流行率为59.38%,而非HCV 1b型慢性感染者、健康对照及慢性HBV感染者中未检测到HCV 1b型F蛋白的抗体,其流行率为0;Western blot方法鉴定了人工合成的HCV 1b型F蛋白抗体的特异性及纯度;ELISA方法检测32例HCV 1b型感染者中有6例检测到HCV 1b型F蛋白抗原的表达(18.75%),而非HCV 1b型慢性感染者、健康对照及慢性HBV感染者中未检测到HCV 1b型F蛋白抗原的表达。结论丙型肝炎病毒F蛋白抗体的产生具有型免疫特异性的特点,不同基因型F蛋白产生的抗体不同,且慢性HCV感染者中有F蛋白抗原的特异性表达。

Objective To study the specific immune response of HCV genotype 1bF protein and assess the prevalence of anti-F antibodies in HCV chronically infected patients. Methods A phenotype ELISA assay was developed to assess the prevalence of specific anti-F antibodies using a specific 99 residue synthetic HCV genotype 1bF peptide.The specific sequence of F peptide was artificially introduced at codon 43 and codon 144as＋1/-1frameshifting from HCV genotype 1bF protein.In order to certify the F protein expression,the specific F antibody was made by mouse immunization and identified by Western blot,and a phenotype ELISA assay was developed using purified antibody to certify the production of HCV 1bF protein antigen and test its prevalence in chronic hepatitis C（CHC）patients. Results The HCV 1bF-protein elicited specific antibodies in 59.38%（19/32）of individuals chronically infected with HCV 1bgenotype using HCV 1bspecific F peptide.No reactivity against HCV 1bspecific F peptide was found in any of the 18non-HCV 1bpatients,20 HBV infected patients and 30 healthy controls.The immunogenicity of the purified anti F antibody was identified by Western blots using HCV 1bspecific F peptide.Interestingly,expression of F protein antigen in 6of 32 HCV 1bpatients were detected and the prevalence rate is about 18.75%. Conclusions The HCV protein bears the genotype immune specific features that different anti-F antibodies were produced by different HCV genotype viruses.The HCV F protein expression was detected in hepatitis C patients and its immunology characteristics in pathogenesis of HCV infected patients need to be further investigated.