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目的分析HCV感染者血清中是否存在交叉反应性中和抗体。方法以分泌表达HCV包膜E2蛋白真核表达质粒转染的293T细胞培养上清液中的HCVE2蛋白作为检测抗原,建立检测HCVE2抗体的ELISA方法,检测32份HCV抗体阳性的慢性丙肝患者血清,然后用免疫荧光分析血清与HCV全长包膜蛋白表达质粒转染的293T细胞的结合反应,再用5株HCV假病毒(HCVpp)及两株细胞培养产生的HCV(HCVcc)为模型分析血清的病毒中和活性。结果 32份HCV抗体阳性血清中,26份血清可检测出E2抗体,阳性率81.3%。其中HCV RNA阳性的12份血清均为E2抗体阳性,E2抗体水平与HCV RNA水平负相关。HCVE2抗体阳性血清对5株HCVpp以及两株HCVcc的感染性均有不同程度的中和作用,中和活性与E2抗体水平相平行。结论 HCV感染可诱导保护性体液免疫应答,丙肝患者血清中存在交叉中和抗体,提示开发能诱导广泛交叉中和抗体的丙肝疫苗具有可行性。
Objective To analyze the presence of a cross-reactive neutralizing antibody in the serum of HCV-infected patients. Methods HCVE2 protein in 293T cell culture supernatant transfected with eukaryotic expression plasmid of HCV envelope E2 protein was used as detection antigen. ELISA method for detecting HCVE2 antibody was established. Serum of 32 HCV-positive chronic hepatitis C patients were detected, Then, the binding of serum to 293T cells transfected with HCV full-length envelope protein expression plasmids was analyzed by immunofluorescence. Then, five strains of HCV pseudovirus (HCVpp) and HCV (HCVcc) produced by two cell cultures were used as a model to analyze serum Virus neutralization activity. Results Of the 32 HCV antibody-positive sera, E2 antibody was detected in 26 sera and the positive rate was 81.3%. Among them, 12 serums positive for HCV RNA were positive for E2 antibody, and E2 antibody level was negatively correlated with HCV RNA level. HCVE2 antibody-positive sera had different degree of neutralization to the infection of 5 HCVpp and 2 HCVcc, and the neutralization activity was in parallel with the E2 antibody level. Conclusion HCV infection can induce a protective humoral immune response. The presence of cross-neutralizing antibodies in the serum of hepatitis C patients suggests that it is feasible to develop a hepatitis C vaccine that can induce a wide range of cross-neutralizing antibodies.