复制型痘病毒载体HIV疫苗免疫策略的系统研究

发布时间：2018-04-06 22:40

本文选题：HIV-1疫苗　切入点：免疫策略　出处：《中国疾病预防控制中心》2017年硕士论文

【摘要】：艾滋病已成为严重威胁世界人民健康的公共卫生问题。安全有效的HIV疫苗可能是控制HIV传播的最佳武器。有关临床试验表明,有效的HIV疫苗可能需要诱导较为平衡的体液免疫和细胞免疫应答。异质的prime—boost免疫策略是目前公认有希望诱导HIV保护性免疫的策略之一。本研究首先基于复制型痘病毒载体疫苗rTV和TTK-EG,对复制型痘病毒载体疫苗初始免疫2次+蛋白疫苗加强免疫2次、DNA疫苗初始免疫3次+复制型痘病毒载体疫苗加强免疫2次、DNA疫苗初始免疫3次+复制型痘病毒载体疫苗/蛋白疫苗联合加强免疫2次3种prime—boost免疫策略进行了研究,并对2次复制型痘病毒载体疫苗单独加强或复制型痘病毒载体疫苗/蛋白疫苗联合加强免疫时不同免疫间隔(免疫间隔为8周、12周或16周)对免疫应答的影响进行了细微研究。实验结果表明,复制型痘病毒载体疫苗rTV初始免疫-蛋白疫苗gp140/p55加强免疫的免疫策略中,免疫间隔对该免疫策略诱导产生的细胞免疫应答无显著性影响,2次rTV之间间隔12周诱导产生的HIV-1 p55抗体有显著增强,抗体滴度几何均数达到105.44,;DNA疫苗初始免疫-复制型痘病毒载体疫苗rTV加强免疫的免疫策略中,2次rTV之间间隔16周能够诱导较高水平的体液免疫和细胞免疫应答,诱导产生的HIV-1V1V2-gp70抗体滴度几何均数达到104.41,分泌IFN-γ、TNF-α的CD8+T细胞比例均值分别为0.86%、1.01%;DNA疫苗初始免疫-复制型痘病毒载体疫苗rTV/蛋白疫苗gp140联合加强免疫的免疫策略中,2次rTV/gp140之间间隔12周可以诱导最为理想的HIV-1特异性体液免疫和细胞免疫应答,诱导产生的HIV-1 gp140抗体滴度的几何均数达到105 93,分泌IFN-丫的CD4+和CD8+ T细胞比例均值分别为0.29%、0.39%。复制型痘病毒载体疫苗TTK-EG初始免疫-蛋白疫苗gp140/p55加强免疫的免疫策略中,2次TTK-EG之间间隔16周诱导产生的HIV-1 V1V2-gp70抗体有显著性增强,抗体滴度几何均数达到104.32;DNA疫苗初始免疫-复制型痘病毒载体疫苗TTK-EG加强免疫的免疫策略中,2次TTK-EG之间隔8周时能够诱导更强的CD4+ T细胞免疫应答,分泌IFN-γ、TNF-α的CD4+ T细胞比例均值分别为0.91%、0.89%,间隔16周时能够诱导更强的CD8+T细胞免疫应答,分泌IFN-γ、TNF-α的CD8+T细胞比例均值分别为0.64%、0.75%;DNA疫苗初始免疫-复制型痘病毒载体疫苗TTK-EG/蛋白疫苗gp140联合加强免疫的免疫策略中,2次TTK-EG/gp140之间间隔16周时诱导产生的HIV-1 V1V2-gp70抗体有显著性提高,抗体滴度几何均数达到10423,同时亦能诱导较强的细胞免疫应答,分泌IFN-γ、TNF-α的CD8+T细胞比例均值分别为0.33%、0.46%。此外本研究进一步对2次TTK-EG肌肉免疫途径所提呈的HIV-1 Env和Gag抗原的优势T细胞表位进行了探索。实验结果表明,经过ELISPOT筛选和ICS验证,确定2条Env多肽(gp140-9,AYDTEVHNVWATHACVPA,氨基酸位点60-77;gp140-74,IVQQQSNLLRAIEAQQHL,氨基酸位点 548-565)和 3 条 Gag多肽(Gag-49,AAMQILKDTINEEAA,氨基酸位点 196-210;Gag-64,VPVGDIYKRWIILGL,氨基酸位点 256-270;Gag-65,DIYKRWIILGLNKIV,氨基酸位点260-274)包含H-2d限制的HIV-1特异性T细胞表位,其中gp140-74(IVQQQSNLLRAIEAQQHL,氨基酸位点548-565)是首次发现的H-2d限制的HIV-1特异性T细胞表位。Env、Gag优势表位肽与Env、Gag全肽池刺激小鼠T细胞分泌IFN-γ、IL-2、TNF-α三种细胞因子的能力基本相同。
[Abstract]:AIDS has become a serious threat to the people of the world public health problem. Safe and effective HIV vaccine may be the best weapon to control the spread of HIV. The clinical trials show that effective HIV vaccine may require humoral and cellular immune responses induced by balanced. The heterogeneity of prime boost immune strategy is regarded as to induce HIV protection immunity strategy one. Based on the type of replication pox virus vector vaccine rTV and TTK-EG on replicative pox virus vector vaccine immune initial 2 + 2 protein vaccine, DNA vaccine immune initial 3 + replicative pox virus vector vaccine 2 times, 3 times the initial DNA vaccine immunization + copy type pox virus vector vaccine / protein vaccine combined with 2 times of 3 prime to strengthen the immune boost immune strategies were studied, and the 2 copy type pox virus vector vaccine alone or strengthen Copy type pox virus vector vaccine / protein vaccine with different immunization immunization (immunization was 8 weeks, 12 weeks or 16 weeks) on immune response to the subtle research. The experimental results show that replicating poxvirus vector rTV vaccine and protein vaccine gp140/ p55 initial immunization strategy to strengthen the immune immunity. No significant effect on cell immune response induced by the interval immunization strategy, HIV-1 p55 antibody between the 2 rTV interval of 12 weeks induced significantly enhanced, geometric mean antibody titer reached 105.44, DNA; initial vaccine immunity - replicating pox virus vector vaccine rTV to strengthen immunization strategies in immunity, humoral immunity and the cellular immune response between the 2 rTV interval of 16 weeks can induce higher levels of antibody titer induced by HIV-1V1V2-gp70, the geometric mean reached 104.41, secreted IFN-, TNF- alpha CD8+T cell ratio The values were 0.86%, 1.01%; the initial DNA vaccine immune - replicating pox virus vector vaccine rTV/ protein vaccine combined with gp140 to strengthen the immune immunity strategy in between the 2 rTV/gp140 interval of 12 weeks most can induce HIV-1 specific humoral and cellular immune responses in the ideal geometry, HIV-1 gp140 antibody titer induced by the mean at 10593, IFN- CD4+ and CD8+ y secretion ratio of T cells which were respectively 0.29%, 0.39%. replication type pox virus vector vaccine TTK-EG initial immune protein vaccine gp140/p55 strengthen the immune immunity strategy in between the 2 TTK-EG interval of 16 weeks of HIV-1 induced by V1V2-gp70 antibody was significantly enhanced, the geometric mean antibody titer reached 104.32; DNA vaccine immune initial replicative poxvirus vector TTK-EG vaccine immunization strategy to strengthen immunity, CD4+ T cells between the 2 TTK-EG every 8 weeks to induce stronger In response, IFN- secretion, TNF- alpha CD4+ T cell mean proportion were 0.91%, 0.89%, CD8+T cell immune response interval of 16 weeks can induce stronger secretion, IFN- gamma, TNF- alpha CD8+T cell mean proportion were 0.64%, 0.75%; the initial DNA vaccine immune - replicating pox virus vector vaccine TTK-EG/ protein vaccine gp140 combined with strengthening immunization strategies in immunity, induced by an interval of 16 weeks when HIV-1 V1V2-gp70 antibody between 2 TTK-EG/gp140 increased significantly and the geometric mean antibody titer reached 10423, at the same time also can induce strong cellular immune response, secreted IFN-, TNF- alpha CD8+T cell mean proportion were 0.33%, in addition to the 0.46%. further research on the 2 TTK-EG the HIV-1 Env muscle immune pathway and Gag antigen were the dominant T cell epitopes were explored. The experimental results show that after ELISPOT selection and ICS verification, to determine the 2 Env polypeptide (G P140-9, AYDTEVHNVWATHACVPA, gp140-74, 60-77 amino acid sites; IVQQQSNLLRAIEAQQHL, 548-565 amino acid sites) and 3 Gag (Gag-49, AAMQILKDTINEEAA, polypeptide of 196-210 amino acid sites; Gag-64, VPVGDIYKRWIILGL, Gag-65, 256-270 amino acid sites; DIYKRWIILGLNKIV, 260-274 amino acid sites) contains H-2d restricted HIV-1 specific T cell epitopes, including gp140-74 (IVQQQSNLLRAIEAQQHL, amino acid site 548-565) is the first discovery of H-2d restricted HIV-1 specific T cell epitope.Env Gag epitope and Env Gag peptide pool stimulated mouse T cells to secrete IFN-, IL-2, TNF- alpha three cytokines are almost the same.

【学位授予单位】：中国疾病预防控制中心
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R392

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