The Epstein-Barr virus membrane antigen was constructed and inserted into vaccinia virus, Tian-tan strain in order to study the effect of this virus on EB infection and tumorogenesis. The EBV-derived membrane antigen ...The Epstein-Barr virus membrane antigen was constructed and inserted into vaccinia virus, Tian-tan strain in order to study the effect of this virus on EB infection and tumorogenesis. The EBV-derived membrane antigen was expressed under the control of a 7.5 K promoter of vaccinia virus. The antibody against the membrane antigen of EB virus was produced on rabbits vaccinated with recombinant vaccinia virus.展开更多
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
This study assessed and compared the immunogenicity of various immunization strategies in mice using combinations of re- combinant DNA （pCCMp24） and recombinant attenuated vaccinia virus Tian Tan （rddVTT_ccMpe4）. ...This study assessed and compared the immunogenicity of various immunization strategies in mice using combinations of re- combinant DNA （pCCMp24） and recombinant attenuated vaccinia virus Tian Tan （rddVTT_ccMpe4）. Intramuscular immuniza- tion was performed on days 0 （prime） and 21 （boost）. The immunogenicity of the vaccine schedules was determined by meas- uring human immunodeficiency virus （HIV）-specific binding antibody levels and cytokine （interleukin-2 and interleukin-4） concentrations in peripheral blood, analyzing lymphocyte proliferation capacity against HIV epitopes and CD4~/CD8＋cell ratio, and monitoring interferon-gamma levels at different times post-immunization. The results showed that pCCMp24, rddVTT.ccMp24 and their prime-boost immunization induced humoral and cellular immune responses. The pCCMp24/ rddVTT.ccMp24 immunization strategy increased CD8＋ T cells and induced more IFN-7-secreting cells compared with sin- gle-shot rDNA. The prime-boost immunization strategy also induced the generation of cellular immunological memory to HIV epitope peptides. These results demonstrated that prime-boost immunization with rDNA and rddVTT_ccMp24 had a tendency to induce greater cellular immune response than single-shot vaccinations, especially IFN-7 response, providing a basis for further studies.展开更多
Conventional influenza vaccines need to be designed and manufactured yearly.However,they occasionally provide poor protection owing to antigenic mismatch.Hence,there is an urgent need to develop universal vaccines aga...Conventional influenza vaccines need to be designed and manufactured yearly.However,they occasionally provide poor protection owing to antigenic mismatch.Hence,there is an urgent need to develop universal vaccines against influenza virus.Using nucleoprotein(NP)and extracellular domain of matrix protein 2(M2e)genes from the influenza A virus A/Beijing/30/95(H3N2),we constructed four recombinant vaccinia virus-based influenza vaccines carrying NP fused with one or four copies of M2e genes in different orders.The recombinant vaccinia viruses were used to immunize BALB/C mice.Humoral and cellular responses were measured,and then the immunized mice were challenged with the influenza A virus A/Puerto Rico/8/34(PR8).NP-specific humoral response was elicited in mice immunized with recombinant vaccinia viruses carrying full-length NP,while robust M2e-specific humoral response was elicited only in the mice immunized with recombinant vaccinia viruses carrying multiple copies of M2e.All recombinant viruses elicited NP-and M2e-specific cellular immune responses in mice.Only immunization with RVJ-4M2eNP induced remarkably higher levels of IL-2 and IL-10 cytokines specific to M2e.Furthermore,RVJ-4M2eNP immunization provided the highest cross-protection in mice challenged with 20 MLD5〇of PR8.Therefore,the cross-protection potentially correlates with both NP and M2e-specific humoral and cellular immune responses induced by RVJ-4M2eNP,which expresses a fusion antigen of full-length NP preceded by four M2e repeats.These results suggest that the rational fusion of NP and multiple M2e antigens is critical toward inducing protective immune responses,and the 4M2eNP fusion antigen may be employed to develop a universal influenza vaccine.展开更多
In order to develop a new vaccine candidate for equine infectious anemia virus (EIAV), gag gene of Chinese donkey leukocyte attenuated strain (EIAV DLV) and its pa- rental virulent strain (EIAV LN) were inserted respe...In order to develop a new vaccine candidate for equine infectious anemia virus (EIAV), gag gene of Chinese donkey leukocyte attenuated strain (EIAV DLV) and its pa- rental virulent strain (EIAV LN) were inserted respectively into the TK region of the Tiantan strain (VV) of vaccinia virus by homologous recombination and the positive clone was confirmed by blue plaque assay. Protein expression was examined by Western blot. Prime and prime-boost proce- dures were used to immunize mice with two DNA vectors and two recombinant vaccinia viruses expressing EIAV Gag pro- teins. The results showed that the specific lysis of CTL re- sponses in the DNA+rVV groups was stronger than those in the DNA groups, amounting to 31%. Although the levels of specific antibodies were not significantly different, we could conclude that the recombinant vaccinia virus could boost the cellular responses following DNA vector priming. There was no detectable difference between the immune responses in- duced by DLV and LN Gag proteins. This data demonstrates that the combined immunity of DNA vector and recombinant vaccinia virus expressing EIAV gag proteins, utilizing the prime-boost procedure, can drive immunized mice to pro- duce powerful cellular responses. These results lay an im- portant foundation for the development of a new EIAV ge- netic engineering vaccine.展开更多
将生长抑素(ss)与乙肝表面抗原(HBsAg)融合基因(ss/HBs)克隆到痘苗病毒表达质粒pGJP-5中,通过与痘苗病毒天坛株体内重组和蚀斑挑选技术获得融合基因的重组病毒,它保持痘苗病毒原有的感染性,并能表达出 ss 和 HBsAg 产物.表达产物呈表面...将生长抑素(ss)与乙肝表面抗原(HBsAg)融合基因(ss/HBs)克隆到痘苗病毒表达质粒pGJP-5中,通过与痘苗病毒天坛株体内重组和蚀斑挑选技术获得融合基因的重组病毒,它保持痘苗病毒原有的感染性,并能表达出 ss 和 HBsAg 产物.表达产物呈表面带 ss 决定簇的 HBsAg 杂合颗粒,并分泌到感染细胞之培养液中.杂合颗粒的大小和密度与报道的 HBsAg 颗粒相似。本文还对该重组痘苗病毒可能作为 ss 活载体苗的前景进行了讨论。展开更多
文摘The Epstein-Barr virus membrane antigen was constructed and inserted into vaccinia virus, Tian-tan strain in order to study the effect of this virus on EB infection and tumorogenesis. The EBV-derived membrane antigen was expressed under the control of a 7.5 K promoter of vaccinia virus. The antibody against the membrane antigen of EB virus was produced on rabbits vaccinated with recombinant vaccinia virus.
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supported by the National Natural Science Foundation of China (81001342)the National Basic Research Program of China (2011CB512110)the National Mega Project on Major Infectious Diseases Prevention (2012ZX10001005-006)
文摘This study assessed and compared the immunogenicity of various immunization strategies in mice using combinations of re- combinant DNA （pCCMp24） and recombinant attenuated vaccinia virus Tian Tan （rddVTT_ccMpe4）. Intramuscular immuniza- tion was performed on days 0 （prime） and 21 （boost）. The immunogenicity of the vaccine schedules was determined by meas- uring human immunodeficiency virus （HIV）-specific binding antibody levels and cytokine （interleukin-2 and interleukin-4） concentrations in peripheral blood, analyzing lymphocyte proliferation capacity against HIV epitopes and CD4~/CD8＋cell ratio, and monitoring interferon-gamma levels at different times post-immunization. The results showed that pCCMp24, rddVTT.ccMp24 and their prime-boost immunization induced humoral and cellular immune responses. The pCCMp24/ rddVTT.ccMp24 immunization strategy increased CD8＋ T cells and induced more IFN-7-secreting cells compared with sin- gle-shot rDNA. The prime-boost immunization strategy also induced the generation of cellular immunological memory to HIV epitope peptides. These results demonstrated that prime-boost immunization with rDNA and rddVTT_ccMp24 had a tendency to induce greater cellular immune response than single-shot vaccinations, especially IFN-7 response, providing a basis for further studies.
基金supported by grant from the National Key Plan for Scientific Research and Development of China (2016YFC1200200)
文摘Conventional influenza vaccines need to be designed and manufactured yearly.However,they occasionally provide poor protection owing to antigenic mismatch.Hence,there is an urgent need to develop universal vaccines against influenza virus.Using nucleoprotein(NP)and extracellular domain of matrix protein 2(M2e)genes from the influenza A virus A/Beijing/30/95(H3N2),we constructed four recombinant vaccinia virus-based influenza vaccines carrying NP fused with one or four copies of M2e genes in different orders.The recombinant vaccinia viruses were used to immunize BALB/C mice.Humoral and cellular responses were measured,and then the immunized mice were challenged with the influenza A virus A/Puerto Rico/8/34(PR8).NP-specific humoral response was elicited in mice immunized with recombinant vaccinia viruses carrying full-length NP,while robust M2e-specific humoral response was elicited only in the mice immunized with recombinant vaccinia viruses carrying multiple copies of M2e.All recombinant viruses elicited NP-and M2e-specific cellular immune responses in mice.Only immunization with RVJ-4M2eNP induced remarkably higher levels of IL-2 and IL-10 cytokines specific to M2e.Furthermore,RVJ-4M2eNP immunization provided the highest cross-protection in mice challenged with 20 MLD5〇of PR8.Therefore,the cross-protection potentially correlates with both NP and M2e-specific humoral and cellular immune responses induced by RVJ-4M2eNP,which expresses a fusion antigen of full-length NP preceded by four M2e repeats.These results suggest that the rational fusion of NP and multiple M2e antigens is critical toward inducing protective immune responses,and the 4M2eNP fusion antigen may be employed to develop a universal influenza vaccine.
文摘In order to develop a new vaccine candidate for equine infectious anemia virus (EIAV), gag gene of Chinese donkey leukocyte attenuated strain (EIAV DLV) and its pa- rental virulent strain (EIAV LN) were inserted respectively into the TK region of the Tiantan strain (VV) of vaccinia virus by homologous recombination and the positive clone was confirmed by blue plaque assay. Protein expression was examined by Western blot. Prime and prime-boost proce- dures were used to immunize mice with two DNA vectors and two recombinant vaccinia viruses expressing EIAV Gag pro- teins. The results showed that the specific lysis of CTL re- sponses in the DNA+rVV groups was stronger than those in the DNA groups, amounting to 31%. Although the levels of specific antibodies were not significantly different, we could conclude that the recombinant vaccinia virus could boost the cellular responses following DNA vector priming. There was no detectable difference between the immune responses in- duced by DLV and LN Gag proteins. This data demonstrates that the combined immunity of DNA vector and recombinant vaccinia virus expressing EIAV gag proteins, utilizing the prime-boost procedure, can drive immunized mice to pro- duce powerful cellular responses. These results lay an im- portant foundation for the development of a new EIAV ge- netic engineering vaccine.
文摘将生长抑素(ss)与乙肝表面抗原(HBsAg)融合基因(ss/HBs)克隆到痘苗病毒表达质粒pGJP-5中,通过与痘苗病毒天坛株体内重组和蚀斑挑选技术获得融合基因的重组病毒,它保持痘苗病毒原有的感染性,并能表达出 ss 和 HBsAg 产物.表达产物呈表面带 ss 决定簇的 HBsAg 杂合颗粒,并分泌到感染细胞之培养液中.杂合颗粒的大小和密度与报道的 HBsAg 颗粒相似。本文还对该重组痘苗病毒可能作为 ss 活载体苗的前景进行了讨论。
