The Immune Effects Of Recombinant Adenovirus Expressing Glycoprotein E Of Varicella-zoster Virus In Different Immunization Routes

Objective:To compare the immune effects of recombinant adenovirus type 63(r Ch Ad63/gE)expressing varicella zoster virus(VZV)gE glycoprotein E(gE)on C57BL/6n mice via intranasal,intradermal and intramuscular immunization routes,so as to lay a foundation for optimizing the immune strategy of herpes zoster adenovirus vector vaccine.Method:First,the eukaryotic expression plasmid p MEW-gE carrying the VZV gE gene was constructed,and the Chinese Hamster Ovary(CHO)cells were transfected with Lipofectamine 2000.After G418 selection screening,stable CHO monoclone cell lines expressing gE protein were obtained.And then gE protein was detected by dot blot and western blot after purification by Ni column affinity chromatography.Subsequently,the titer of recombinant adenovirus r Ch Ad63/gE was measured by immunospot method and Qubit fluorescent quantification.C57BL/6n mice were immunized by intranasal,intradermal and intramuscular immunization routes.Spleen cells were isolated after serum collection.The titer of gE specific serum antibody was detected by ELISA,and the specific CD4~+T cell and CD8~+T cell immune responses were detected by flow cytometry.Results:A eukaryotic expression vector p MEW was constructed and a stable CHO cell line expression of VZV gE protein was generated.And gE protein with a concentration of 348.65μg/m L was obtained after Ni column affinity chromatography purification for subsequent experiments.Compared to the intramuscular injection group,intranasal immunization of C57BL/6n mice with r Ch Ad63/gE induced a similar level of CD4~+T and CD8~+T cell response secreting IFN-γ,CD4~+T and CD8~+T cell response secreting IFN-γand IL-2,CD8~+T cell response secreting IL-2,yet induced a higher level of CD4~+T cell response and higher humoral immune response secreting of IL-2.Compared to the intramuscular injection group,intradermal immunization of C57BL/6n mice with r Ch Ad63/gE induced similar levels of CD4~+T and CD8~+T cell response.Conclusion:(1)A stable CHO cell line expresseing VZV gE protein was successfully established,and the gE protein was expressed and purified(2)C57BL/6n mice immunized with recombinant adenovirus via intranasal and intradermal routes can induce good humoral and cellular immune responses,which is an appropriate immune pathway for adenovirus based herpes zoster vaccine.