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  本文關(guān)鍵詞：炭疽桿菌保護(hù)性抗原重組腺病毒疫苗的構(gòu)建及氣溶膠免疫探索性研究　出處：《中國人民解放軍軍事醫(yī)學(xué)科學(xué)院》2006年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 炭疽桿菌 保護(hù)性抗原 重組腺病毒 氣溶膠

【摘要】：目的：探討利用腺病毒載體作為炭疽桿菌基因工程疫苗載體的可行性；探索重組腺病毒對霧化的耐受能力；探討腺病毒活載體疫苗霧化吸入免疫途徑的可行性。 方法： 1．PCR擴增PA基因后克隆至質(zhì)粒pAdTrack-CMV，經(jīng)與pAdeasy-1同源重組，構(gòu)建重組腺病毒vAd-PA，轉(zhuǎn)染293細(xì)胞，經(jīng)Western-blot檢測表明PA在293細(xì)胞中得到表達(dá)。 2．重組腺病毒肌肉注射和滴鼻免疫途徑免疫BALB／c小鼠，通過ELISA和MTT等實驗對免疫鼠的特異性抗體、黏膜免疫反應(yīng)及細(xì)胞免疫應(yīng)答進(jìn)行分析。 3．優(yōu)化重組腺病毒霧化條件，選擇最優(yōu)條件霧化免疫小鼠，通過ELISA和MTT等實驗對免疫鼠的特異性抗體、黏膜免疫反應(yīng)及細(xì)胞免疫應(yīng)答進(jìn)行分析。 結(jié)果： 1．通過酶切鑒定的方法驗證，成功構(gòu)建了重組腺病毒vAd-PA，并用Western-blot的方法驗證了PA蛋白在細(xì)胞內(nèi)的表達(dá)。 2．重組腺病毒通過肌肉注射和滴鼻的途徑免疫小鼠，均能誘導(dǎo)特異的體液、黏膜和細(xì)胞免疫應(yīng)答。表現(xiàn)為血清中IgG升高，肺灌洗液中的IgA升高，以及脾細(xì)胞培養(yǎng)上清中的細(xì)胞因子濃度有所升高，MMT實驗結(jié)果顯示T細(xì)胞增殖程度高于對照組。 3．重組腺病毒霧化途徑免疫小鼠，可誘導(dǎo)特異的體液、黏膜和細(xì)胞免疫應(yīng)答。表現(xiàn)為血清中IgG升高，肺灌洗液中的IgA升高，MMT實驗結(jié)果顯示T細(xì)胞增殖程度高于對照組。 結(jié)論：炭疽桿菌重組腺病毒基因工程疫苗有望發(fā)展成為一種新型的基因工程疫苗；霧化吸入免疫途徑可考慮作為炭疽桿菌腺病毒活載體疫苗的免疫方式。
[Abstract]:Objective: to explore the feasibility of using adenovirus vector as gene engineering vaccine vector of Bacillus anthracis. To explore the tolerance of recombinant adenovirus to atomization; To investigate the feasibility of aerosol inhalation of adenovirus live vector vaccine. Methods: 1. The PA gene was amplified by PCR and cloned into plasmid pAdTrack-CMV. After homologous recombination with pAdeasy-1, the recombinant adenovirus vAd-PA was constructed. After transfection into 293 cells, Western-blot analysis showed that PA was expressed in 293 cells. 2. BALB/c mice were immunized by recombinant adenovirus intramuscular injection and nasal drip immunization. Specific antibodies were obtained by ELISA and MTT. Mucosal immune response and cellular immune response were analyzed. 3. Optimize the nebulization conditions of recombinant adenovirus and select the optimal conditions to immunize mice. The specific antibodies of immunized mice were obtained by ELISA and MTT experiments. Mucosal immune response and cellular immune response were analyzed. Results: 1. The recombinant adenovirus vAd-PAwas successfully constructed by restriction endonuclease assay, and the expression of PA protein in the cells was verified by Western-blot. 2. The recombinant adenovirus could induce specific humoral, mucosal and cellular immune responses in mice by intramuscular injection and nasal drip. IgA in lung lavage fluid and cytokine concentration in the supernatant of spleen cell culture were increased. The results showed that the proliferation of T cells was higher than that of control group. 3. The specific humoral, mucosal and cellular immune responses were induced by recombinant adenovirus nebulization pathway in mice. The results showed that IgG in serum and IgA in lung lavage fluid were increased. MMT results showed that T cell proliferation was higher than that in control group. Conclusion: the recombinant adenovirus gene engineering vaccine of Bacillus anthracis is expected to be a new type of genetic engineering vaccine. Nebulized inhalation can be considered as a vaccine against anthrax adenovirus.
【學(xué)位授予單位】：中國人民解放軍軍事醫(yī)學(xué)科學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2006
【分類號】：R392.1

【引證文獻(xiàn)】

相關(guān)博士學(xué)位論文 前1條

1 于龍;模式病毒（噬菌體）分離、特性及在防護(hù)裝備和設(shè)施評價中的應(yīng)用研究[D];中國人民解放軍軍事醫(yī)學(xué)科學(xué)院;2010年

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本文編號：1413140