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[A bivalent VP1 gene vaccine against Coxsackie virus B1/B3]
Objective:
To construct a bivalent VP1 gene vaccine against Coxsackie virus B1 and B3 (CVB1 and CVB3) and to test its immunogenicity in mice.
Methods:
The VP1 gene of CVB1 was amplified by RT-PCR. The VP1 genes of CVB1 and of CVB3 under the control of cytomegalovirus (CMV) promoter and SV40 promoter respectively were inserted oppositely into the pCR3-Uni, a eukaryotic expression vector. The resultant bivalent gene vaccine plasmid was used to inoculate 3-week-old BALB/c mice. Immunofluorescence test and RT-PCR were conducted 24, 48, and 96 hours after the transfection. Sera of the tested animals were sampled at 0, 2, 4, and 6 weeks after the inoculation and detected for CVB-specific antibodies by ELISA.
Results:
A bivalent gene vaccine plasmid of CVB1 and CVB3 was obtained. Samples of 4 and 6 weeks after the inoculation were shown to be CVB-antibody positive, while those of 0 and 2 weeks after the inoculation shown negative.
Conclusion:
The gene vaccine thus constructed induces CVB-speicific antibodies in mice, and can serve as gene vaccine candidate for human beings.