發(fā)布時(shí)間：2018-04-06 22:40

  本文選題：HIV-1疫苗　切入點(diǎn)：免疫策略　出處：《中國疾病預(yù)防控制中心》2017年碩士論文

【摘要】：艾滋病已成為嚴(yán)重威脅世界人民健康的公共衛(wèi)生問題。安全有效的HIV疫苗可能是控制HIV傳播的最佳武器。有關(guān)臨床試驗(yàn)表明,有效的HIV疫苗可能需要誘導(dǎo)較為平衡的體液免疫和細(xì)胞免疫應(yīng)答。異質(zhì)的prime—boost免疫策略是目前公認(rèn)有希望誘導(dǎo)HIV保護(hù)性免疫的策略之一。本研究首先基于復(fù)制型痘病毒載體疫苗rTV和TTK-EG,對復(fù)制型痘病毒載體疫苗初始免疫2次+蛋白疫苗加強(qiáng)免疫2次、DNA疫苗初始免疫3次+復(fù)制型痘病毒載體疫苗加強(qiáng)免疫2次、DNA疫苗初始免疫3次+復(fù)制型痘病毒載體疫苗/蛋白疫苗聯(lián)合加強(qiáng)免疫2次3種prime—boost免疫策略進(jìn)行了研究,并對2次復(fù)制型痘病毒載體疫苗單獨(dú)加強(qiáng)或復(fù)制型痘病毒載體疫苗/蛋白疫苗聯(lián)合加強(qiáng)免疫時(shí)不同免疫間隔(免疫間隔為8周、12周或16周)對免疫應(yīng)答的影響進(jìn)行了細(xì)微研究。實(shí)驗(yàn)結(jié)果表明,復(fù)制型痘病毒載體疫苗rTV初始免疫-蛋白疫苗gp140/p55加強(qiáng)免疫的免疫策略中,免疫間隔對該免疫策略誘導(dǎo)產(chǎn)生的細(xì)胞免疫應(yīng)答無顯著性影響,2次rTV之間間隔12周誘導(dǎo)產(chǎn)生的HIV-1 p55抗體有顯著增強(qiáng),抗體滴度幾何均數(shù)達(dá)到105.44,;DNA疫苗初始免疫-復(fù)制型痘病毒載體疫苗rTV加強(qiáng)免疫的免疫策略中,2次rTV之間間隔16周能夠誘導(dǎo)較高水平的體液免疫和細(xì)胞免疫應(yīng)答,誘導(dǎo)產(chǎn)生的HIV-1V1V2-gp70抗體滴度幾何均數(shù)達(dá)到104.41,分泌IFN-γ、TNF-α的CD8+T細(xì)胞比例均值分別為0.86%、1.01%;DNA疫苗初始免疫-復(fù)制型痘病毒載體疫苗rTV/蛋白疫苗gp140聯(lián)合加強(qiáng)免疫的免疫策略中,2次rTV/gp140之間間隔12周可以誘導(dǎo)最為理想的HIV-1特異性體液免疫和細(xì)胞免疫應(yīng)答,誘導(dǎo)產(chǎn)生的HIV-1 gp140抗體滴度的幾何均數(shù)達(dá)到105 93,分泌IFN-丫的CD4+和CD8+ T細(xì)胞比例均值分別為0.29%、0.39%。復(fù)制型痘病毒載體疫苗TTK-EG初始免疫-蛋白疫苗gp140/p55加強(qiáng)免疫的免疫策略中,2次TTK-EG之間間隔16周誘導(dǎo)產(chǎn)生的HIV-1 V1V2-gp70抗體有顯著性增強(qiáng),抗體滴度幾何均數(shù)達(dá)到104.32;DNA疫苗初始免疫-復(fù)制型痘病毒載體疫苗TTK-EG加強(qiáng)免疫的免疫策略中,2次TTK-EG之間隔8周時(shí)能夠誘導(dǎo)更強(qiáng)的CD4+ T細(xì)胞免疫應(yīng)答,分泌IFN-γ、TNF-α的CD4+ T細(xì)胞比例均值分別為0.91%、0.89%,間隔16周時(shí)能夠誘導(dǎo)更強(qiáng)的CD8+T細(xì)胞免疫應(yīng)答,分泌IFN-γ、TNF-α的CD8+T細(xì)胞比例均值分別為0.64%、0.75%;DNA疫苗初始免疫-復(fù)制型痘病毒載體疫苗TTK-EG/蛋白疫苗gp140聯(lián)合加強(qiáng)免疫的免疫策略中,2次TTK-EG/gp140之間間隔16周時(shí)誘導(dǎo)產(chǎn)生的HIV-1 V1V2-gp70抗體有顯著性提高,抗體滴度幾何均數(shù)達(dá)到10423,同時(shí)亦能誘導(dǎo)較強(qiáng)的細(xì)胞免疫應(yīng)答,分泌IFN-γ、TNF-α的CD8+T細(xì)胞比例均值分別為0.33%、0.46%。此外本研究進(jìn)一步對2次TTK-EG肌肉免疫途徑所提呈的HIV-1 Env和Gag抗原的優(yōu)勢T細(xì)胞表位進(jìn)行了探索。實(shí)驗(yàn)結(jié)果表明,經(jīng)過ELISPOT篩選和ICS驗(yàn)證,確定2條Env多肽(gp140-9,AYDTEVHNVWATHACVPA,氨基酸位點(diǎn)60-77;gp140-74,IVQQQSNLLRAIEAQQHL,氨基酸位點(diǎn) 548-565)和 3 條 Gag多肽(Gag-49,AAMQILKDTINEEAA,氨基酸位點(diǎn) 196-210;Gag-64,VPVGDIYKRWIILGL,氨基酸位點(diǎn) 256-270;Gag-65,DIYKRWIILGLNKIV,氨基酸位點(diǎn)260-274)包含H-2d限制的HIV-1特異性T細(xì)胞表位,其中g(shù)p140-74(IVQQQSNLLRAIEAQQHL,氨基酸位點(diǎn)548-565)是首次發(fā)現(xiàn)的H-2d限制的HIV-1特異性T細(xì)胞表位。Env、Gag優(yōu)勢表位肽與Env、Gag全肽池刺激小鼠T細(xì)胞分泌IFN-γ、IL-2、TNF-α三種細(xì)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.

【學(xué)位授予單位】：中國疾病預(yù)防控制中心
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R392

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 齊香榮;高瑛瑛;陸柔劍;鄧瑤;孟昕;譚文杰;阮力;;應(yīng)用ELISPOT方法篩選確定HIV-1 B'/C亞型疫苗六種抗原的H-2~d限制的T細(xì)胞表位[J];病毒學(xué)報(bào);2011年01期

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本文編號(hào)：1719264