The vaccinia virus Tiantan(VTT)is widely utilized as a smallpox vaccine in China and holds significant impor-tance in the prevention of diseases stemming from poxvirus infections.Nevertheless,few studies have investi-...The vaccinia virus Tiantan(VTT)is widely utilized as a smallpox vaccine in China and holds significant impor-tance in the prevention of diseases stemming from poxvirus infections.Nevertheless,few studies have investi-gated the influence of VTT infection on host gene expression.In this study,we constructed time series transcriptomic profiles of HeLa cells infected with both VTT and western reserve(WR)strains.We observed similar patterns of viral gene expression,while the expression levels of host genes varied between the two strains.There was an immediate and significant repression of host gene expression,particularly in genes asso-ciated with oxidative phosphorylation.Conversely,genes involved in nerve growth factor(NGF)-stimulated transcription were significantly activated.The upregulation of genes linked to the ribonucleic acid(RNA)-induced silencing complex(RISC)suggested a potential role for posttranscriptional regulation in the interac-tion between the vaccinia virus and the host.In the later stages of infection,pathways such as extracellular matrix organization,neutrophil degranulation,complement and interferon responses,translation,and pro-grammed cell death are largely inhibited.A significant number of host genes exhibit correlations with changes in the expression levels of viral genes.The host genes that are negatively correlated with viral genes are mainly enriched in pathways associated with translation and the response to viral infection.This study significantly contributes to advancing our understanding of the dynamics between the vaccinia virus and the host,improv-ing the application of VTTs and facilitating the development of effective vaccines against diseases such as smallpox and monkeypox.展开更多
Background Although DNA vaccine is considered as the next generation of vaccine, most DNA vaccine candidates are still suffering from the relatively weak immunogenicity despite the increased dosage of plasmid DNA admi...Background Although DNA vaccine is considered as the next generation of vaccine, most DNA vaccine candidates are still suffering from the relatively weak immunogenicity despite the increased dosage of plasmid DNA administered. In order to enhance the immune responses elicited by a codon-optimized HIV gag DNA vaccine, a modified plasmid vector pDRVI1.0 and a booster immunization with replicating Tiantan vaccinia （RTV） strain expressing the same gene were employed. Methods Vector pDRVI1.0 was constructed through inserting the 72-bp element from the SV40 enhancer, which was reported promoting nuclear transport of plasmid DNA, to the upstream of cytomegalovirus enhancer/promoter region of the plasmid vector pVR1012. Gene expression levels from expression plasmids based on pDRVI1.0 and pVR1012 were tested. Humoral and cellular immune responses induced by DNA vaccine alone or DNA prime-RTV boost regimen were determined in mice. Results It was shown that the 72-bp element significantly enhanced the gene expression level in non-dividing cells. gag-specific humoral and cellular immune responses induced by DNA vaccination were both significantly improved, while the Thl/Th2 balance was not obviously affected by the 72-bp element. RTV boosting further significantly enhanced DNA vaccine-palmed antibody and T cell responses in a Thl-biased manner. Conclusions The 72-bp SV40 enhancer element should be included in the DNA vaccine vector and RTV strain is a very efficient live vector for boosting immunization.展开更多
基金supported by the National Key Research and Development Program of China(2022YFC2303401)the National Natural Science Foundation of China(82241066).
文摘The vaccinia virus Tiantan(VTT)is widely utilized as a smallpox vaccine in China and holds significant impor-tance in the prevention of diseases stemming from poxvirus infections.Nevertheless,few studies have investi-gated the influence of VTT infection on host gene expression.In this study,we constructed time series transcriptomic profiles of HeLa cells infected with both VTT and western reserve(WR)strains.We observed similar patterns of viral gene expression,while the expression levels of host genes varied between the two strains.There was an immediate and significant repression of host gene expression,particularly in genes asso-ciated with oxidative phosphorylation.Conversely,genes involved in nerve growth factor(NGF)-stimulated transcription were significantly activated.The upregulation of genes linked to the ribonucleic acid(RNA)-induced silencing complex(RISC)suggested a potential role for posttranscriptional regulation in the interac-tion between the vaccinia virus and the host.In the later stages of infection,pathways such as extracellular matrix organization,neutrophil degranulation,complement and interferon responses,translation,and pro-grammed cell death are largely inhibited.A significant number of host genes exhibit correlations with changes in the expression levels of viral genes.The host genes that are negatively correlated with viral genes are mainly enriched in pathways associated with translation and the response to viral infection.This study significantly contributes to advancing our understanding of the dynamics between the vaccinia virus and the host,improv-ing the application of VTTs and facilitating the development of effective vaccines against diseases such as smallpox and monkeypox.
基金This project was supported by the CIPRA Program granted by the National Institute of Health(No.1 U19 AI51915-02)a grant from the National High Technology Research and Development Program of China(863 Program,No.2003AA219100)
文摘Background Although DNA vaccine is considered as the next generation of vaccine, most DNA vaccine candidates are still suffering from the relatively weak immunogenicity despite the increased dosage of plasmid DNA administered. In order to enhance the immune responses elicited by a codon-optimized HIV gag DNA vaccine, a modified plasmid vector pDRVI1.0 and a booster immunization with replicating Tiantan vaccinia （RTV） strain expressing the same gene were employed. Methods Vector pDRVI1.0 was constructed through inserting the 72-bp element from the SV40 enhancer, which was reported promoting nuclear transport of plasmid DNA, to the upstream of cytomegalovirus enhancer/promoter region of the plasmid vector pVR1012. Gene expression levels from expression plasmids based on pDRVI1.0 and pVR1012 were tested. Humoral and cellular immune responses induced by DNA vaccine alone or DNA prime-RTV boost regimen were determined in mice. Results It was shown that the 72-bp element significantly enhanced the gene expression level in non-dividing cells. gag-specific humoral and cellular immune responses induced by DNA vaccination were both significantly improved, while the Thl/Th2 balance was not obviously affected by the 72-bp element. RTV boosting further significantly enhanced DNA vaccine-palmed antibody and T cell responses in a Thl-biased manner. Conclusions The 72-bp SV40 enhancer element should be included in the DNA vaccine vector and RTV strain is a very efficient live vector for boosting immunization.
