Antigen Gene Cloning and Expression of HIV-1 for AIDS Vaccine Design Ⅲ. HIV-1 Antigen Gene Cloning, in Vitro Expression and Antibody Induction

Objective: To evaluate the humoral immune induction in rats of a candidate AIDS vaccine expressing the gag p24 gene froma subtype B HIV-1 isolate. Methods: The amplified p24 gene was inserted into aneukaryotic expression vector to form the supercoiled DNAvaccine. The linearized expressed DNA vaccine was preparedfrom the expression plasmid by polymerase chain reaction(PCR). The antigen gene expression in rats of the linearizedand supercoiled DNA vaccines were in vitro and in vivodetected. Results: In vitro transcription and Northern hybridizationshowed that the linearized DNA vaccine could synthesizeamounts or p24 mRNA similar to the supercoiled DNA vaccine.Antibody assays of inoculated rats confirmed that thelinearized expression DNA could induce a slightly higherantibody titer than the expression plasmid, while the highestautibody titer had been induced by plasmid plus adjuvantinoculation. Conclusion: The construction of a candidate AIDS vaccinebased on the p24 gene could shed light on a potential IV vaccine, meriting evaluation in a rhesus macaque SHIV-AIDSmodel.

Objective: To evaluate the humoral immune induction in rats of a candidate AIDS vaccine expressing the gag p24 gene from a subtype B HIV-1 isolate. Methods: The amplified p24 gene was inserted into an eukaryotic expression vector to form the supercoiled DNA vaccine. The linearized expressed DNA vaccine was preparedfrom the expression plasmid by polymerase chain reaction(PCR). The antigen gene expression in rats of the linearized and supercoiled DNA vaccines were in vitro and in vivodetected. Results: In vitro transcription and Northern hybridizationshowed that the linearized DNA vaccine could synthesizeamounts of p24 mRNA similar to the supercoiled DNA vaccine.Antibody assays of inoculated rats confirmed that thelinearized expression DNA could induce a slightly higherantibody titer than the expression plasmid, while the highestantibody titer had been induced by plasmid plus adjuvantinoculation. Conclusion: The construction of a candidate AIDS vaccinebased on the p24 gene could shed light on a potential HIVvaccine, meriting evaluation in a rhesus macaque SHIV-AIDSmodel.