本文選題：艾滋病疫苗　切入點(diǎn)：佐劑　出處：《北京工業(yè)大學(xué)》2014年碩士論文

【摘要】：研制安全有效的艾滋病疫苗控制人類免疫缺陷病毒（HIV）傳播是遏制艾滋病流行的最好方法。雖然已有幾十種疫苗進(jìn)行了臨床試驗(yàn)，但是還沒(méi)有一種疫苗對(duì)高危人群具有有效的保護(hù)作用。繼續(xù)進(jìn)行新型艾滋病疫苗研發(fā)刻不容緩，艾滋病治療性疫苗能夠在較長(zhǎng)的時(shí)間內(nèi)控制病毒載量、延緩艾滋病發(fā)病進(jìn)程，為艾滋病疫苗研發(fā)的主要方向，，同時(shí)利用各種佐劑增強(qiáng)艾滋病疫苗的免疫效力也是目前研究中的新策略。 本研究首先對(duì)本課題組前期構(gòu)建的表達(dá)HIV-1gp160的腺病毒5型載體（Ad5-HIVgp160）疫苗進(jìn)行臨床前小鼠藥效學(xué)研究。Ad5-HIVgp160疫苗以2×106VP/只、2×107VP/只、2×108VP/只、2×109VP/只、2×1010VP/只劑量分別免疫小鼠，采用ELISPOT法和ELISA法檢測(cè)免疫小鼠體內(nèi)特異性細(xì)胞免疫反應(yīng)和體液免疫反應(yīng)，結(jié)果各劑量組小鼠均可誘導(dǎo)出特異性細(xì)胞免疫反應(yīng)和體液免疫反應(yīng)，劑量為2×108VP/只時(shí)能夠在小鼠體內(nèi)激發(fā)出高水平的細(xì)胞免疫反應(yīng)和體液免疫反應(yīng)。 之后，研究IL15基因佐劑協(xié)同poly(I:C)對(duì)表達(dá)HIV-1B亞型gp160DNA及腺病毒載體疫苗免疫增強(qiáng)作用，并探討其作用機(jī)制。首先構(gòu)建表達(dá)IL15的質(zhì)粒pVR-IL15，將pVR-IL15聯(lián)合pVR-HIVgp160初免Ad5-HIVgp160加強(qiáng)方式免疫小鼠，采用IFN-γ ELISPOT、ELISA和CCK-8等方法比較疫苗單獨(dú)免疫組小鼠與疫苗加IL15佐劑組小鼠誘導(dǎo)的細(xì)胞免疫、體液免疫反應(yīng)及淋巴細(xì)胞增殖反應(yīng)的強(qiáng)度。結(jié)果顯示疫苗加IL15佐劑組小鼠誘導(dǎo)的細(xì)胞免疫反應(yīng)、體液免疫反應(yīng)及淋巴細(xì)胞增殖反應(yīng)比疫苗單獨(dú)免疫組小鼠相比均有顯著增強(qiáng)。pVR-IL15協(xié)同poly(I:C)聯(lián)合pVR-HIVgp160初免Ad5-HIVgp160加強(qiáng)方式免疫小鼠，同樣采用上述方法檢測(cè)免疫小鼠體內(nèi)特異性細(xì)胞免疫反應(yīng)和淋巴細(xì)胞增殖反應(yīng)，結(jié)果顯示pVR-IL15協(xié)同poly(I:C)組小鼠特異性細(xì)胞免疫反應(yīng)及淋巴細(xì)胞增殖反應(yīng)均顯著高于單獨(dú)使用pVR-IL15組小鼠。進(jìn)一步實(shí)驗(yàn)說(shuō)明，pVR-IL15協(xié)同poly(I:C)組小鼠在免疫后3天即可誘導(dǎo)出高水平的特異性細(xì)胞免疫反應(yīng)，高于其它各組小鼠，并且平穩(wěn)保持至免疫后第8周。熒光定量PCR實(shí)驗(yàn)進(jìn)一步顯示，pVR-IL15協(xié)同poly(I:C)組小鼠淋巴細(xì)胞中Th1型細(xì)胞因子IFN-γ及抗凋亡因子Bcl-2的表達(dá)均高于其他各組小鼠，并且Bcl-2表達(dá)量與特異性細(xì)胞免疫反應(yīng)及淋巴細(xì)胞增殖反應(yīng)強(qiáng)度呈一定相關(guān)性。 綜上所述，Ad5-HIVgp160疫苗可以誘導(dǎo)機(jī)體產(chǎn)生特異性細(xì)胞免疫應(yīng)答和體液免疫應(yīng)答，有可能成為有效的艾滋病治療性疫苗；IL15基因佐劑協(xié)同poly(I:C)可以增強(qiáng)HIV DNA疫苗初免/腺病毒疫苗加強(qiáng)免疫策略的免疫效果，延長(zhǎng)免疫反應(yīng)作用時(shí)間，為IL15佐劑協(xié)同poly(I:C)臨床應(yīng)用提供實(shí)驗(yàn)支持。
[Abstract]:The development of a safe and effective AIDS vaccine to control the spread of human immunodeficiency virus (HIV) is the best way to contain the AIDS epidemic. Although dozens of vaccines have been tested in clinical trials, However, none of the vaccines has an effective protective effect on high-risk groups. It is urgent to continue the development of new AIDS vaccines. AIDS therapeutic vaccines can control the load of the virus in a longer period of time and delay the onset of AIDS. The main direction of AIDS vaccine research and development, and the use of various adjuvants to enhance the immune effectiveness of AIDS vaccine is also a new strategy in current research. In this study, we first studied the preclinical pharmacodynamics of Ad5-HIVgp160 (Ad5-HIVgp160) vaccine constructed by our research group. Ad5-HIVgp160 vaccine was immunized with 2 脳 106VP/, 2 脳 107VP/, 2 脳 108VP/, 2 脳 109VP/ and 2 脳 1010VP/, respectively. The specific cellular and humoral immune responses of immunized mice were detected by ELISPOT and ELISA methods. The results showed that specific cellular and humoral immune responses could be induced in each dose group of mice. When the dose of 2 脳 108VP/ was 2 脳 108VP/ only, the high level of cellular and humoral immune response could be stimulated in mice. After that, the effects of IL15 gene adjuvant combined with polysid I: C on the expression of HIV-1B subtype gp160DNA and adenovirus vector vaccine were studied, and the mechanism was discussed. Firstly, the plasmid pVR-IL15 expressing IL15 was constructed, and the combination of pVR-IL15 and pVR-HIVgp160 was used to immunize mice with Ad5-HIVgp160. IFN- 緯 Elispot Elisa and CCK-8 were used to compare the cellular immunity induced by vaccine alone and IL15 adjuvant. The results showed that the immune response of mice induced by vaccine and IL15 adjuvant was induced by humoral immune reaction and lymphocyte proliferation. The humoral immune response and lymphocyte proliferation reaction were significantly enhanced compared with the mice immunized with vaccine alone. PVR-IL15 combined with pVR-HIVgp160 combined with Ad5-HIVgp160 was used to immunize mice. The above methods were also used to detect the specific cellular immune response and lymphocyte proliferation in immunized mice. The results showed that the specific cellular immune response and lymphocyte proliferation of mice in pVR-IL15 combined with polysid I: C group were significantly higher than those in mice treated with pVR-IL15 alone. Further experiments showed that the mice in the group of pVR-IL15 combined with polycyclic I: C) could induce high water content 3 days after immunization. The specific cellular immune response, The expression of Th1 type cytokine IFN- 緯 and anti-apoptotic factor (Bcl-2) in the lymphocytes of the mice treated with pVR-IL15 combined with polygonal I: C- (1) was higher than that of the other groups, and the expression of IFN- 緯 and anti-apoptotic factor Bcl-2 in the lymphocytes of the mice with pVR-IL15 combined with polymorphic I: C was higher than that of the other groups. The expression of Bcl-2 was correlated with the specific cellular immune response and lymphocyte proliferation. In conclusion, Ad5-HIVgp160 vaccine can induce specific cellular and humoral immune responses. It is possible to be an effective adjuvant of IL15 gene for AIDS treatment vaccine combined with poly (I: C). It can enhance the immune effect of HIV DNA vaccine primary immunization / adenovirus vaccine and prolong the time of immune response. To provide experimental support for clinical application of IL15 adjuvant in collaboration with polyacrylamide I: C.
【學(xué)位授予單位】：北京工業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R392

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 馮霞;楊海儒;余雙慶;周玲;李紅霞;李澤琳;曾毅;;河南省有償供血者HIV-1外膜蛋白env基因序列分析及表型預(yù)測(cè)[J];病毒學(xué)報(bào);2009年02期

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