Development of poliovirus-rotavirus chimeric vaccine by using poliovirus artificial interfering particle RNA

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We constructed polio-rota VP4 chimeric cDNA by inserting the DNA fragments encoding VP8' or VP5' into pSM1 (T7) 3 or pSM1 (T7) 2 vector having poliovirus artificial DI genome. In the HeLa cells transfected with polio-rota chimeric RNA obtained from pSM1 (T7) 3-VP5', pSM1 (T7) 3-VP8', or pSM1 (T7) 2-VP8'+VP5', replication of rotavirus-specific RNA could be detected. However, the expression of rotaviral protein was observed only in the cells transfected with pSM1 (T7) 3-VP8' or pSM1 (T7) 2-VP8'+VP5'. The antigenicity of the expressed VP5' might be weak in this system because of the conformational change in the from of the VP5' fused with polioviral protein. Rotaviral protein reactive with antiserum to VP4 could be detected in the cells at the fourth generation after transfection with pSM1(T7)3-VP8' and infection with infectious poiliovirus. By preparing of polio-rota chimeric virus with high titer and by using the transgenic mice with poliovirus receptor which have the potency of oral inoculation, the immunogenicity of the poliorota chimeric virus would be investigated.We determined the nucleotide and deduced amino acid sequences of the VP4 genes of the rotaviruses from various mammalian species including humans. Comparative analysis of the VP4 sequences showed that there are at least 17 kinds of VP4 genotypes and that species specificity and interspecies relatedness in VP4 geneotypes are present. It was also shown that some human rotaviruses have VP4 geneotypes related to those of animal rotaviruses. These findings should be taken into considerataion for rotavirus vaccine development.