汉滩病毒包膜糖蛋白嵌合DNA疫苗的细胞免疫评价及其优势表位的筛选
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摘要
目的:筛选Gn和Gc抗原优势表位,为汉滩病毒疫苗研究提供方向.方法:通过构建汉坦病毒包膜糖蛋白Gn与Gc嵌合溶酶体相关膜蛋白的DNA疫苗pVAX-LAMP/Gn,pVAX-LAMP/Gc,利用酶联免疫斑点实验(ELISpot)长期评估了BALB/c小鼠体内的细胞免疫应答.结果:在刺激T淋巴细胞的短效与长效免疫中,pVAX-LAMP/Gc组的效应优于其他组,并筛选出优势抗原表位.结论:目前许多新型汉滩病毒疫苗研究缺少细胞免疫应答评价和记忆免疫应答评价.筛选出的Gn和Gc的抗原优势表位,可以为新型汉滩病毒疫苗的研制奠定良好的基础.
AIM: To screen the dominant epitopes of antigens and provide a direction for further research of Hantaanvirus vaccine. METHODS: DNA vaccine encoding Hantavirus glycoproteins fused with lysosome-associated membrane protein was designed in this study. cellular immune response of BALB/c mice was assessed by ELISpot. RESULTS: The p VAX-LAMP/Gc group showed great advantages in stimulating short-term and longterm immunity reaction, and dominant antigen epitopes were found. CONCLUSION: Currently,lots of HFRS vaccine researches is limitted in cellular immune response and memory immune response evaluation. Epitope mapping was performed to study the T-cell epitopes. This study lays a good foundation for the research of Hantavirus vaccine.
AIM: To screen the dominant epitopes of antigens and provide a direction for further research of Hantaanvirus vaccine. METHODS: DNA vaccine encoding Hantavirus glycoproteins fused with lysosome-associated membrane protein was designed in this study. cellular immune response of BALB/c mice was assessed by ELISpot. RESULTS: The p VAX-LAMP/Gc group showed great advantages in stimulating short-term and longterm immunity reaction, and dominant antigen epitopes were found. CONCLUSION: Currently,lots of HFRS vaccine researches is limitted in cellular immune response and memory immune response evaluation. Epitope mapping was performed to study the T-cell epitopes. This study lays a good foundation for the research of Hantavirus vaccine.
引文
[1]Av2cic-6upanc T,Saksida A,Korva M.Hantavirus infections[J].Clin Microbiol Infect,2015.
[2]Mir MA.Hantaviruses[J].Clin Lab Med,2010,30(1):67-91.
[3]Jiang H,Zheng X,Wang L,et al.Hantavirus infection:a global zoonotic challenge[J].Virol Sin,2017,32(1):32-43.
[4]SzabóR.Antiviral therapy and prevention against hantavirus infections[J].Acta Virol,2017,61(1):3-12.
[5]Battisti AJ,Chu YK,Chipman PR,et al.Structural studies of Hantaan virus[J].J Virol,2011,85(2):835-841.
[6]Muyangwa M,Martynova EV,Khaiboullina SF,et al.Hantaviral Proteins:Structure,Functions,and Role in Hantavirus Infection[J].Front Microbiol,2015,6:1326.
[7]Hooper JW,Josleyn M,Ballantyne J,et al.A novel Sin Nombre virus DNA vaccine and its inclusion in a candidate pan-hantavirus vaccine against hantavirus pulmonary syndrome(HPS)and hemorrhagic fever with renal syndrome(HFRS)[J].Vaccine,2013,31(40):4314-4321.
[8]Higa MM,Petersen J,Hooper J,et al.Efficient production of Hantaan and Puumala pseudovirions for viral tropism and neutralization studies[J].Virology,2012,423(2):134-142.
[9]Marques ET Jr,Chikhlikar P,de Arruda LB,et al.HIV-1 p55Gag encoded in the lysosome-associated membrane protein-1 as a DNAplasmid vaccine chimera is highly expressed,traffics to the major histocompatibility classⅡcompartment,and elicits enhanced immune responses[J].J Biol Chem,2003,278(39):37926-37936.
[10]Anwar A,Chandrasekaran A,Ng ML,et al.West Nile premembrane-envelope genetic vaccine encoded as a chimera containing the transmembrane and cytoplasmic domains of a lysosome-associated membrane protein:increased cellular concentration of the transgene product,targeting to the MHC II compartment,and enhanced neutralizing antibody response[J].Virology,2005,332(1):66-77.
[11]Chikhlikar P,Barros de Arruda L,Maciel M,et al.DNA encoding an HIV-1 Gag/human lysosome-associated membrane protein-1chimera elicits a broad cellular and humoral immune response in Rhesus macaques[J].PLo S One,2006,1:e135.
[12]Nakanishi Y,Lu B,Gerard C,et al.CD8(+)T lymphocyte mobilization to virus-infected tissue requires CD4(+)T-cell help[J].Nature,2009,462(7272):510-513.
[13]Hooper JW,Li D.Vaccines against hantaviruses[J].Curr Top Microbiol Immunol,2001,256:171-191.
[14]Boudreau EF,Josleyn M,Ullman D,et al.A Phase 1 clinical trial of Hantaan virus and Puumala virus M-segment DNA vaccines for hemorrhagic fever with renal syndrome[J].Vaccine,2012,30(11):1951-1958.
[15]Hooper JW,Moon JE,Paolino KM,et al.A Phase 1 clinical trial of Hantaan virus and Puumala virus M-segment DNA vaccines for haemorrhagic fever with renal syndrome delivered by intramuscular electroporation[J].Clin Microbiol Infect,2014,20Suppl 5:110-117.
[16]Song JY,Woo HJ,Cheong HJ,et al.Long-term immunogenicity and safety of inactivated Hantaan virus vaccine(HantavaxTM)in healthy adults[J].Vaccine,2016,34(10):1289-1295.
[17]Ma Y,Wang J,Yuan B,et al.HLA-A2 and B35 restricted hantaan virus nucleoprotein CD8+T-cell epitope-specific immune response correlates with milder disease in hemorrhagic fever with renal syndrome[J].PLo S Negl Trop Dis,2013,7(2):e2076.
[18]Ma Y,Yuan B,Zhuang R,et al.Hantaan virus infection induces both Th1 and Th Granzyme B+cell immune responses that associated with viral control and clinical outcome in humans[J].PLo S Pathog,2015,11(4):e1004788.
[19]Yang K,Sun K,Srinivasan KN,et al.Immune responses to T-cell epitopes of SARS Co V-N protein are enhanced by N immunization with a chimera of lysosome-associated membrane protein[J].Gene Ther,2009,16(11):1353-1362.
[20]Dhalia R,Maciel MJr,Cruz FS,et al.Membrane and envelope virus proteins co-expressed as lysosome associated membrane protein(LAMP)fused antigens:a potential tool to develop DNA vaccines against flaviviruses[J].An Acad Bras Cienc,2009,81(4):663-669.
[21]Hussein IT,Cheng E,Ganaie SS,et al.Autophagic clearance of Sin Nombre hantavirus glycoprotein Gn promotes virus replication in cells[J].J Virol,2012,86(14):7520-7529.
[22]Ganaie SS,Mir MA.The role of viral genomic RNA and nucleocapsid protein in the autophagic clearance of hantavirus glycoprotein Gn[J].Virus Res,2014,187:72-76.
[23]Ma RX,Cheng LF,Ying QK,et al.Screening and Identification of an H-2Kb-Restricted CTL epitope within the glycoprotein of hantaan virus[J].Front Cell Infect Microbiol,2016,6:151.
[2]Mir MA.Hantaviruses[J].Clin Lab Med,2010,30(1):67-91.
[3]Jiang H,Zheng X,Wang L,et al.Hantavirus infection:a global zoonotic challenge[J].Virol Sin,2017,32(1):32-43.
[4]SzabóR.Antiviral therapy and prevention against hantavirus infections[J].Acta Virol,2017,61(1):3-12.
[5]Battisti AJ,Chu YK,Chipman PR,et al.Structural studies of Hantaan virus[J].J Virol,2011,85(2):835-841.
[6]Muyangwa M,Martynova EV,Khaiboullina SF,et al.Hantaviral Proteins:Structure,Functions,and Role in Hantavirus Infection[J].Front Microbiol,2015,6:1326.
[7]Hooper JW,Josleyn M,Ballantyne J,et al.A novel Sin Nombre virus DNA vaccine and its inclusion in a candidate pan-hantavirus vaccine against hantavirus pulmonary syndrome(HPS)and hemorrhagic fever with renal syndrome(HFRS)[J].Vaccine,2013,31(40):4314-4321.
[8]Higa MM,Petersen J,Hooper J,et al.Efficient production of Hantaan and Puumala pseudovirions for viral tropism and neutralization studies[J].Virology,2012,423(2):134-142.
[9]Marques ET Jr,Chikhlikar P,de Arruda LB,et al.HIV-1 p55Gag encoded in the lysosome-associated membrane protein-1 as a DNAplasmid vaccine chimera is highly expressed,traffics to the major histocompatibility classⅡcompartment,and elicits enhanced immune responses[J].J Biol Chem,2003,278(39):37926-37936.
[10]Anwar A,Chandrasekaran A,Ng ML,et al.West Nile premembrane-envelope genetic vaccine encoded as a chimera containing the transmembrane and cytoplasmic domains of a lysosome-associated membrane protein:increased cellular concentration of the transgene product,targeting to the MHC II compartment,and enhanced neutralizing antibody response[J].Virology,2005,332(1):66-77.
[11]Chikhlikar P,Barros de Arruda L,Maciel M,et al.DNA encoding an HIV-1 Gag/human lysosome-associated membrane protein-1chimera elicits a broad cellular and humoral immune response in Rhesus macaques[J].PLo S One,2006,1:e135.
[12]Nakanishi Y,Lu B,Gerard C,et al.CD8(+)T lymphocyte mobilization to virus-infected tissue requires CD4(+)T-cell help[J].Nature,2009,462(7272):510-513.
[13]Hooper JW,Li D.Vaccines against hantaviruses[J].Curr Top Microbiol Immunol,2001,256:171-191.
[14]Boudreau EF,Josleyn M,Ullman D,et al.A Phase 1 clinical trial of Hantaan virus and Puumala virus M-segment DNA vaccines for hemorrhagic fever with renal syndrome[J].Vaccine,2012,30(11):1951-1958.
[15]Hooper JW,Moon JE,Paolino KM,et al.A Phase 1 clinical trial of Hantaan virus and Puumala virus M-segment DNA vaccines for haemorrhagic fever with renal syndrome delivered by intramuscular electroporation[J].Clin Microbiol Infect,2014,20Suppl 5:110-117.
[16]Song JY,Woo HJ,Cheong HJ,et al.Long-term immunogenicity and safety of inactivated Hantaan virus vaccine(HantavaxTM)in healthy adults[J].Vaccine,2016,34(10):1289-1295.
[17]Ma Y,Wang J,Yuan B,et al.HLA-A2 and B35 restricted hantaan virus nucleoprotein CD8+T-cell epitope-specific immune response correlates with milder disease in hemorrhagic fever with renal syndrome[J].PLo S Negl Trop Dis,2013,7(2):e2076.
[18]Ma Y,Yuan B,Zhuang R,et al.Hantaan virus infection induces both Th1 and Th Granzyme B+cell immune responses that associated with viral control and clinical outcome in humans[J].PLo S Pathog,2015,11(4):e1004788.
[19]Yang K,Sun K,Srinivasan KN,et al.Immune responses to T-cell epitopes of SARS Co V-N protein are enhanced by N immunization with a chimera of lysosome-associated membrane protein[J].Gene Ther,2009,16(11):1353-1362.
[20]Dhalia R,Maciel MJr,Cruz FS,et al.Membrane and envelope virus proteins co-expressed as lysosome associated membrane protein(LAMP)fused antigens:a potential tool to develop DNA vaccines against flaviviruses[J].An Acad Bras Cienc,2009,81(4):663-669.
[21]Hussein IT,Cheng E,Ganaie SS,et al.Autophagic clearance of Sin Nombre hantavirus glycoprotein Gn promotes virus replication in cells[J].J Virol,2012,86(14):7520-7529.
[22]Ganaie SS,Mir MA.The role of viral genomic RNA and nucleocapsid protein in the autophagic clearance of hantavirus glycoprotein Gn[J].Virus Res,2014,187:72-76.
[23]Ma RX,Cheng LF,Ying QK,et al.Screening and Identification of an H-2Kb-Restricted CTL epitope within the glycoprotein of hantaan virus[J].Front Cell Infect Microbiol,2016,6:151.