RESUMO
A lamb strain of rotavirus has recently been licensed for use in China as a live vaccine to prevent rotavirus diarrhea in children. As rotavirus NSP4, especially the cytotoxic domain alone is considered to be associated with diarrhea, we sequenced gene segment 10, which encodes NSP4, of lamb rotavirus. Comparative analyses was performed to identify differences from human rotavirus strains, that might be associated with attenuation, and to ascertain whether the lamb rotavirus gene fits among the NSP4 of other sequenced rotavirus strains. Our comparative nucleotide sequence analysis suggests its close identity (91.17% homology) with that of group-A equine rotavirus (strain HI23). Multiple alignment of the deduced amino acid sequence of lamb NSP4 with that of other group A rotaviruses demonstrated homology ranging from 63.42% with that of porcine YM strain to 93.71% with equine HI23 strain of rotavirus. A group A-specific NSP4 monoclonal antibody recognized the glycosylated and unglycosylated forms of the protein from virus-infected lysates, suggesting a well-conserved group-specificity of the lamb NSP4. Phylogenetic analysis of the lamb rotavirus gene, with 60 other NSP4 gene sequences of human and animal rotavirus strains, demonstrated that the lamb rotavirus strain belongs to genotype A. Comparative analysis also revealed that although it is a vaccine strain, the NSP4 cytotoxic domain of lamb strain demonstrated an overall amino acid conservation similar to that of other strains, whose NSP4 alone causes diarrhea in animal models. These results taken together with our previous observations clearly reaffirm the idea that the attenuation phenotype of rotaviruses does not involve NSP4 cytotoxic domain, perhaps due to the suppression of NSP4 cytotoxic activity by other rotaviral proteins.
Assuntos
Genes Virais , Vacinas contra Rotavirus/genética , Rotavirus/genética , Rotavirus/imunologia , Ovinos/virologia , Sequência de Aminoácidos , Animais , Sequência de Bases , Criança , China , DNA Viral/genética , Diarreia/prevenção & controle , Humanos , Dados de Sequência Molecular , Filogenia , Rotavirus/classificação , Infecções por Rotavirus/prevenção & controle , Infecções por Rotavirus/virologia , Homologia de Sequência de Aminoácidos , Homologia de Sequência do Ácido NucleicoRESUMO
Objective:To discuss mice cellular immune response to the hepatitis C viral nucleic acid vaccine.Methods:Hepatitis C viral nucleic acid vaccine pSVL HCV/C+E 1 was inoculated BALB/C mice by subcutaneous injection.To detect the CTL activity of immune mice,Made the standard 4 hours lysis test by labeling the target cells with (Na) 2 51 CrO 4,which was prepared from the syngentic BALB/C mouse myeloma derived cell strain SP2/0 transfected by the eukaryotic expression recombinant plasmid pEGFP N 1 HCV/C+E 1.Results:The kill ratio of CTL is up to 40.7%.Conclusion:The result suggested hepatitis C viral nucleic acid vaccine could induce strong cellular immune response
