Ⅱ型脊灰病毒抗原表位嵌合蛋白的免疫学研究

Immunological Activity of a Chimeric Protein Carrying an Epitope of Type 2 Poliovirus on the VP6 of Rotavirus as a Vector

目的:评价以轮状病毒(RV)重组VP6蛋白为载体插入Ⅱ型脊髓灰质炎病毒(PV2)VP1蛋白上的1个抗原表位构建而成的嵌合蛋白的体外免疫学性质。方法:采用分子克隆和基因重组技术将PV2抗原表位插入到RV载体蛋白上,在大肠杆菌中表达并用SDS-PAGE确认表达产物,再通过动物免疫、Western blot、免疫荧光和病毒血清抗体中和试验分析嵌合蛋白的免疫学性质。结果:成功构建了以VP6为载体的PV2抗原表位嵌合蛋白6F/PV2N1,并且在E.coli系统中高效表达,嵌合蛋白免疫的豚鼠血清抗体对RV和PV2具备较好的中和活性。结论:以RV VP6为载体构建的嵌合蛋白具有较好的免疫原性,免疫豚鼠产生血清抗体可中和RV和PV2在体外细胞上的感染;进一步为研发RV/PV2嵌合疫苗提供了较好的基础。

Objective：To evaluate immunological activity of chimeric protein carrying one PV2 antigenic epitope on rotavirus VP6. Methods：one chimeric protein was constructed by inserting an epitope derived from neutralizing antigenic sites 1 of PV2 in rotavirus VP6 as a vector through gene cloning and recombination; the chimeric protein was expressed in E. coli cells and detected by Western blot, immunofluorescence assay and neutralization test. Results： The chimeric protein was favorably constructed and expressed in E. coli cells efficiently. Antibodies induced in inoculated guinea pigs by the chimeric protein could both neutralize the infection of RV and PV2 in vitro. Conclusion：The constructed chimeric protein possesses dual immunogenicity and can elicit production of antibodies in immunized guinea pigs which can protect infection of RV in MA104 cells and PV in Vero cells. The results are valuable for development of new RV/PV2 bivalent vaccine.