E.coli表达HEV ORF2编码蛋白第112～607氨基酸片段的免疫原性及其诱生抗体的病毒中和活性

Immunogenicity of Amino Acids 112-607 of A Recombinant HEV ORF2-Encoded Protein Expressed in E. cott and Neutralizing Capacity of Antibodies Elicited by Its Immunization in Animals

目的 探讨E.coli表达的戊型肝炎病毒(HEV)ORF2编码蛋白第112～607氨基酸片段(pORF2-56K)的免疫原性及其诱生抗体的病毒中和活性。方法 pORF2-56K免疫小鼠和豚鼠,用ELISA检测血清抗体效价。将HEV毒种与高效价豚鼠灭活血清在体外孵育后,接种于2BS细胞,用RT-PCR检测病毒RNA的复制情况。结果E.coli表达的pORF2-56K可诱导机体产生高水平的HEV特异性抗体和强烈的免疫记忆。该抗体可在体外有效中和HEV,使之失去在2BS细胞中有效复制的能力。结论 E-coli表达的pORF2-56K具有良好的免疫原性,且其诱生抗体具有中和HEV的能力,可作为HEV重组疫苗的候选抗原。

Objective To explore the immunogenicity of pORF2-56K,a recombinant HEV ORF2-en-coded protein (aminp acids 112-607) expressed in E. coli, as well as the neutralizing capacity of antibodies induced with it. Methods Immunize mice and guinea pigs with pORF2-56K and detect serum antibody liters by ELISA. Incubate HEV strain with inactivated guinea pig sera at high liters, inoculate into 2BS cells and detect the replication of viral RNA by RT-PCR. Results The pORF2-56K expressed in E. coli induced a high HEV-specific antibody level and a strong immune memory.The antibody neutralized HEV in vitro and prevented it from replication in 2BS cells. Conclusion The pORF2-56K expressed in E. coll showed good immunogenicity , and the antibody induced with it showed neutralizing capacity to HEV. So pORF2-56K might be used as a candidate antigen for the development of a recombinant HEV vaccine.