DNA疫苗-重组痘苗病毒-蛋白疫苗联合诱导HIV-1抗体应答免疫程序的优化
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摘要
比较DNA疫苗、重组痘苗病毒、蛋白疫苗不同免疫组合及免疫间隔时间对免疫效果的影响。采取Primeboost-boost免疫程序对豚鼠进行DNA疫苗初免三次,重组痘苗病毒rTV加强一次,分别间隔4、8、12周后免疫gp140蛋白加强免疫两次;同时,采取Prime-boost免疫程序重组痘苗病毒rTV初免一次,分别间隔4、8、12周后加强免疫gp140蛋白二次。末次免疫后不同时间点采血检测豚鼠血清中HIV-1特异性结合抗体、中和抗体以及抗体的相对亲合力。DNA-rTV-gp140免疫策略诱导的HIV-1gp140/gp120抗体滴度、相对亲合力以及V1V2-gp70抗体OD值均高于rTV-gp140。对DNA-rTV-gp140免疫策略而言,间隔12周的免疫策略较间隔4周诱导了更高的V1V2-gp70抗体,而间隔4周更有利于提高抗体亲合力。rTV-gp140免疫策略间隔8或12周诱导的体液免疫更为理想。DNA疫苗初免,rTV和蛋白疫苗依次加强能够诱导更强的体液免疫,较长的免疫间隔有利于诱导V1V2-gp70抗体,较短的间隔有利于提高抗体亲合力。
To optimize the immunization strategy against HIV-1,a DNA vaccine was combined with a recombinant vaccinia virus(rTV)vaccine and a protein vaccine.Immune responses against HIV-1were detected in 30 female guinea pigs divided into six groups.Three groups of guinea pigs were primed with HIV-1DNA vaccine three times,boosted with rTV at week 14,and then boosted with gp140 protein at intervals of 4,8or 12 weeks.Simultaneously,the other three groups of animals were primed with rTV vaccine once,and then boosted with gp140 after 4,8or 12 weeks.The HIV-1specific binding antibody and neutralizing antibody,in addition to the relative affinity of these antibodies,were detected at different time points after the final administration of vaccine in each group.The DNA-rTV-gp140 immune regimen induced higher titers and affinity levels of HIV-1gp120/gp140 antibodies and stronger V1V2-gp70 antibodies than the rTV-gp140 regimen.In the guinea pigs that underwent the DNA-rTV-gp140 regimen,the highest V1V2-gp70 antibody was induced in the 12-week-interval group.However,the avidity of antibodies was improved in the 4-week-interval group.Using the rTV-gp140 immunization strategy,guinea pigs boosted at 8or 12 weeks after rTV priming elicited stronger humoral responses than those boosted at 4weeks after priming.In conclusion,this study shows that the immunization strategy of HIV-1DNA vaccine priming,followed by rTV and protein vaccine boosting,could strengthen the humoral response against HIV-1.Longer intervals were better to induce V1V2-gp70-specific antibodies,while shorter intervals were more beneficial to enhance the avidity of antibodies.
To optimize the immunization strategy against HIV-1,a DNA vaccine was combined with a recombinant vaccinia virus(rTV)vaccine and a protein vaccine.Immune responses against HIV-1were detected in 30 female guinea pigs divided into six groups.Three groups of guinea pigs were primed with HIV-1DNA vaccine three times,boosted with rTV at week 14,and then boosted with gp140 protein at intervals of 4,8or 12 weeks.Simultaneously,the other three groups of animals were primed with rTV vaccine once,and then boosted with gp140 after 4,8or 12 weeks.The HIV-1specific binding antibody and neutralizing antibody,in addition to the relative affinity of these antibodies,were detected at different time points after the final administration of vaccine in each group.The DNA-rTV-gp140 immune regimen induced higher titers and affinity levels of HIV-1gp120/gp140 antibodies and stronger V1V2-gp70 antibodies than the rTV-gp140 regimen.In the guinea pigs that underwent the DNA-rTV-gp140 regimen,the highest V1V2-gp70 antibody was induced in the 12-week-interval group.However,the avidity of antibodies was improved in the 4-week-interval group.Using the rTV-gp140 immunization strategy,guinea pigs boosted at 8or 12 weeks after rTV priming elicited stronger humoral responses than those boosted at 4weeks after priming.In conclusion,this study shows that the immunization strategy of HIV-1DNA vaccine priming,followed by rTV and protein vaccine boosting,could strengthen the humoral response against HIV-1.Longer intervals were better to induce V1V2-gp70-specific antibodies,while shorter intervals were more beneficial to enhance the avidity of antibodies.
引文
[1]Berman P W,Gregory T J,Riddle L,et al.Protection of chimpanzees from infection by HIV-1after vaccination with recombinant glycoprotein gp120but not gp160[J].Nature,1990Jun 14;345(6276):p.622-625.
[2]el-Amad Z,Murthy K K,Hiqqins K,et al.Resistance of chimpanzees immunized with recombinant gp120SF2to challenge by HIV-1SF2[J].AIDS,1995 Dec;9(12):p.1313-1322.
[3]阎冬梅.HIV候选疫苗不同免疫策略下免疫效果的比较研究[D].佳木斯:佳木斯大学,2005年7月.
[4]Hutnick N A,Myles D J,Bian C B,et al.Selected approaches for increasing HIV DNA vaccine immunogencty in vivo[J].Curr Opin Virol,2011Oct;1(4):233-240.
[5]Mwan M,Cebere I,Sutton J,et al.A human immunodeficiency virus 1(HIV-1)clade A vaccine in clinical trials:stimulation of HIV-specific T-cell responses by DNA and recombinant modified vaccinia virus Ankara(MVA)vaccines in humans[J].J Gen Virol,2004 Apr;85(Pt4):911-919.
[6]Doria-Rose N A,Ohlen C,Polacino P,et a1.Multigene DNA priming-boosting vaccines protect macaques from acute CD4+-T-ceII depletion after simian-human immunodeficiency virus SHIV89.6P mucosal challenge[J].J Virol,2003Nov,77(21):11563-11577.
[7]Leung L,Srivastava I K,Kan E,et al.Immunogencity of HIV-1Env and Gag in baboons using a DNA prime/protein boost regimen[J].AIDS,2004Apr 30;18(7):991-1001.
[8]Zolla-Pazner S,Kong X P,Jiang X,et al.Cross-clade HIV-1neutralizing antibodies induced with V3-scaffold protein immunogens following priming with gp120DNA[J].J Virol,2011Oct;85(19):9887-9898.
[9]Gorelick R J,Benveniste R E,Lifson J D,et al.Protection of Macacs nemestrina from disease following pathogenic simian immunodeficiency virus(SIV)challenge:utilization of SIV nucleocapsid mutant DNA vaccines with and without an SIV protein boost[J].J Virol,2000Dec;74(24):11935-11949.
[10]Cherpelis S,Shrivastava I,Gettie A,et al.DNA vaccination with the human immunodeficiency virus type 1SF162DeltaV2envelop elicits immune responses that offer partial protection from simian/human immunodeficiency virus infection to CD8(+)T-cell-depleted rhesus macaques[J].J Virol,2001Feb;75(3):1547-1550.
[11]张舟,齐智,刘颖,等.HIV-1疫苗prime-boost策略免疫间隔研究[J].中国热带医学2013,13:393-397.
[12]刘颖,段丹丽,彭虹,等.HIV复制型痘苗病毒载体疫苗免疫原性分析[J].中华实验临床病毒学杂志,2004,18(3):281-283.
[13]Dai K,Liu Y,Liu M,et al.Pathogenicity and immunogenicity of recombinant Tiantan Vaccinia Virus with deleted C12Land A53R genes[J].Vacine,2008Sep15;26(39):5062-5071.
[14]林怡.HIV-1中和抗体的作用与机制及实验室检测进展[J].中国艾滋病性病,2008,14:634-636.
[15]Horton H,Vogel T U,Carter D K,et al.Immunization of rhesus macaques with a DNA prime/modified vaccina virus Ankara boost regimen induces broad simian immunodeficiency virus(SIV)-specific T-cell responses and reduces intial viral replication but dose not prevent disease progression following challenge with pathogenic SIVmac239[J].J Virol,2002Jul;76(14):p.7187-7202.
[16]Amara R R,Smith J M,Staprans S I,et al.Critical role for Env as well as Gag-Pol in control of a simian-human immunodeficiency virus 89.6P challenge by a DNA prime/recombinant modified vaccinia virus Ankara vaccine[J].J Virol,2002Jun;76(12):p.6138-6146.
[17]Amara R R,Sharma S,Patel M,et al.Studies on the cross-clade and cross-species conservation of HIV-1Gag-specific CD8and CD4Tcell responses elicited by a clade B DNA/MVA vaccine in macaques[J].Virology,2005 Mar 30;334(1):p.124-133.
[18]Haynes B F,Gilbert P B,McElrath M J,et al.Immunecorrelates analysis of an HIV-1vaccine efficacy trial[J].N Engl J Med,2012,Apr 5;366(14):1275-1286.
[19]Zolla-Pazner S,deCamp A,Gilbert P B,et al.Vaccineinduced IgG antibodies to V1V2regions of multiple HIV-1subtypes correlate with decreased risk of HIV-1infection[J/OL].PLoS One,2014Feb 4;9(2):e87572.
[20]Doria-Rose N A,Schramm C A,Gorman J,et al.Developmental pathway for potent V1V2-directed HIV-neutralizing antibodies[J].Nature,2014 May 1;509(7498):55-62.
[21]Bontjer I,Melchers M,Tong T,et al.Comparative immunogenicity of evolved V1V2-deleted HIV-1envelope glycoprotein trimers[J/OL].PLoS One,2013Jun 26;8(6):e67484.
[2]el-Amad Z,Murthy K K,Hiqqins K,et al.Resistance of chimpanzees immunized with recombinant gp120SF2to challenge by HIV-1SF2[J].AIDS,1995 Dec;9(12):p.1313-1322.
[3]阎冬梅.HIV候选疫苗不同免疫策略下免疫效果的比较研究[D].佳木斯:佳木斯大学,2005年7月.
[4]Hutnick N A,Myles D J,Bian C B,et al.Selected approaches for increasing HIV DNA vaccine immunogencty in vivo[J].Curr Opin Virol,2011Oct;1(4):233-240.
[5]Mwan M,Cebere I,Sutton J,et al.A human immunodeficiency virus 1(HIV-1)clade A vaccine in clinical trials:stimulation of HIV-specific T-cell responses by DNA and recombinant modified vaccinia virus Ankara(MVA)vaccines in humans[J].J Gen Virol,2004 Apr;85(Pt4):911-919.
[6]Doria-Rose N A,Ohlen C,Polacino P,et a1.Multigene DNA priming-boosting vaccines protect macaques from acute CD4+-T-ceII depletion after simian-human immunodeficiency virus SHIV89.6P mucosal challenge[J].J Virol,2003Nov,77(21):11563-11577.
[7]Leung L,Srivastava I K,Kan E,et al.Immunogencity of HIV-1Env and Gag in baboons using a DNA prime/protein boost regimen[J].AIDS,2004Apr 30;18(7):991-1001.
[8]Zolla-Pazner S,Kong X P,Jiang X,et al.Cross-clade HIV-1neutralizing antibodies induced with V3-scaffold protein immunogens following priming with gp120DNA[J].J Virol,2011Oct;85(19):9887-9898.
[9]Gorelick R J,Benveniste R E,Lifson J D,et al.Protection of Macacs nemestrina from disease following pathogenic simian immunodeficiency virus(SIV)challenge:utilization of SIV nucleocapsid mutant DNA vaccines with and without an SIV protein boost[J].J Virol,2000Dec;74(24):11935-11949.
[10]Cherpelis S,Shrivastava I,Gettie A,et al.DNA vaccination with the human immunodeficiency virus type 1SF162DeltaV2envelop elicits immune responses that offer partial protection from simian/human immunodeficiency virus infection to CD8(+)T-cell-depleted rhesus macaques[J].J Virol,2001Feb;75(3):1547-1550.
[11]张舟,齐智,刘颖,等.HIV-1疫苗prime-boost策略免疫间隔研究[J].中国热带医学2013,13:393-397.
[12]刘颖,段丹丽,彭虹,等.HIV复制型痘苗病毒载体疫苗免疫原性分析[J].中华实验临床病毒学杂志,2004,18(3):281-283.
[13]Dai K,Liu Y,Liu M,et al.Pathogenicity and immunogenicity of recombinant Tiantan Vaccinia Virus with deleted C12Land A53R genes[J].Vacine,2008Sep15;26(39):5062-5071.
[14]林怡.HIV-1中和抗体的作用与机制及实验室检测进展[J].中国艾滋病性病,2008,14:634-636.
[15]Horton H,Vogel T U,Carter D K,et al.Immunization of rhesus macaques with a DNA prime/modified vaccina virus Ankara boost regimen induces broad simian immunodeficiency virus(SIV)-specific T-cell responses and reduces intial viral replication but dose not prevent disease progression following challenge with pathogenic SIVmac239[J].J Virol,2002Jul;76(14):p.7187-7202.
[16]Amara R R,Smith J M,Staprans S I,et al.Critical role for Env as well as Gag-Pol in control of a simian-human immunodeficiency virus 89.6P challenge by a DNA prime/recombinant modified vaccinia virus Ankara vaccine[J].J Virol,2002Jun;76(12):p.6138-6146.
[17]Amara R R,Sharma S,Patel M,et al.Studies on the cross-clade and cross-species conservation of HIV-1Gag-specific CD8and CD4Tcell responses elicited by a clade B DNA/MVA vaccine in macaques[J].Virology,2005 Mar 30;334(1):p.124-133.
[18]Haynes B F,Gilbert P B,McElrath M J,et al.Immunecorrelates analysis of an HIV-1vaccine efficacy trial[J].N Engl J Med,2012,Apr 5;366(14):1275-1286.
[19]Zolla-Pazner S,deCamp A,Gilbert P B,et al.Vaccineinduced IgG antibodies to V1V2regions of multiple HIV-1subtypes correlate with decreased risk of HIV-1infection[J/OL].PLoS One,2014Feb 4;9(2):e87572.
[20]Doria-Rose N A,Schramm C A,Gorman J,et al.Developmental pathway for potent V1V2-directed HIV-neutralizing antibodies[J].Nature,2014 May 1;509(7498):55-62.
[21]Bontjer I,Melchers M,Tong T,et al.Comparative immunogenicity of evolved V1V2-deleted HIV-1envelope glycoprotein trimers[J/OL].PLoS One,2013Jun 26;8(6):e67484.