發(fā)布時間：2018-05-12 00:31

  本文選題：吸附PA苗 + A16R　； 參考：《中國人民解放軍軍事醫(yī)學(xué)科學(xué)院》2007年碩士論文

【摘要】： 炭疽芽孢桿菌是人畜共患炭疽病的重要病原體，是公認(rèn)的重要生物戰(zhàn)劑之一，目前人用疫苗有兩種：即炭疽減毒活芽孢苗和炭疽弱毒株無菌培養(yǎng)濾液佐劑吸附苗，自研制成功以來，已在人群中長期使用，為了深入了解炭疽疫苗的免疫應(yīng)答機(jī)制，為疫苗的開發(fā)及評價提供理論基礎(chǔ)，本研究采用炭疽芽孢桿菌兩種疫苗——A16R芽孢苗與吸附PA苗，，分別從三個層面上——脾、骨髓和外周血，初步觀察炭疽兩種疫苗的免疫應(yīng)答機(jī)制。 本研究共分為三部分：1、炭疽兩種疫苗的制備及免疫效果測定，分兩部分(1)兩種炭疽疫苗的制備和鑒定；(2)A16R芽孢苗及吸附PA苗免疫Balb／C小鼠血清抗體效價動態(tài)觀察及抗體亞型分析；2、A16R芽孢苗及吸附PA苗誘導(dǎo)Balb／C小鼠特異性ASCs的動態(tài)觀察(1)首先建立了ELISPOT檢測炭疽疫苗免疫Balb／C小鼠后抗原特異性抗體分泌細(xì)胞的實(shí)驗(yàn)方法；(2)從三個層面上，即骨髓、脾和外周血，觀察了兩種疫苗免疫Balb／C小鼠后特異性ASCs的動態(tài)變化及差異性，并結(jié)合外周血抗體效價的消長分析不同器官／部位ASCs應(yīng)答與外周血抗體效價消長之間的相關(guān)性；3、炭疽A16R芽孢苗與吸附PA苗誘導(dǎo)Balb／C小鼠脾、骨髓及外周血中特異性Th2細(xì)胞應(yīng)答的動態(tài)變化，并結(jié)合外周血抗體效價的消長分析不同器官／部位Th2應(yīng)答的產(chǎn)生與維持對穩(wěn)定血中抗芽孢／PA抗體的輔助作用。 結(jié)論：(1)通過對炭疽兩種疫苗免疫后Balb／C小鼠外周血抗體效價的動態(tài)觀察，發(fā)現(xiàn)隨著兩種疫苗的加強(qiáng)免疫，小鼠血清中均可檢測到高效價的抗體水平，并維持較長時間；吸附PA苗免疫組抗體亞型分析以IgG1和IgG2b為主，IgG2a水平較低，與對照組差異顯著；A16R芽孢苗免疫組抗體亞型分析雖仍以IgG1和IgG2b為主，亦可檢測到較高水平的IgG2a；(2)兩種疫苗免疫后Balb／C小鼠脾、骨髓和血中均可檢測到ASCs，但ASCs出現(xiàn)時間及強(qiáng)度不同，脾中最先出現(xiàn)，首先達(dá)峰值，骨髓中峰值晚于脾，但骨髓中應(yīng)答維持時間較長，與外周血抗體效價的維持有一定的相關(guān)性；兩種疫苗免疫后Balb／C小鼠脾、骨髓及外周血中ASCs的動態(tài)變化有一定差異；(3)兩種疫苗免疫后Balb／C小鼠脾、骨髓和外周血中均可檢測到抗原特異性Th2細(xì)胞應(yīng)答，但Th2應(yīng)答出現(xiàn)時間及強(qiáng)度不同，骨髓中最先出現(xiàn)，首先達(dá)峰值，且維持時間長，外周血中可檢測到較強(qiáng)的Th2應(yīng)答，且隨著脾、骨髓及外周血的Th2應(yīng)答的升高與維持，外周血抗體滴度不斷升高，且于末次加強(qiáng)免疫后60天或30天仍維持在較高水平。疫苗免疫應(yīng)答是一個復(fù)雜的多層次的過程，受復(fù)雜的網(wǎng)絡(luò)調(diào)控，因此對于疫苗的評價應(yīng)從多方面、多角度入手，全面、深入、系統(tǒng)的研究。
[Abstract]:Bacillus anthracis is an important pathogen of zoonotic anthracnose and one of the most important biological warfare agents. At present, there are two kinds of human vaccines: anthrax attenuated live spores vaccine and anthrax attenuated strain sterile culture filtrate adjuvant adsorbent vaccine. Since the successful development of anthrax vaccine, it has been used for a long time. In order to understand the immune response mechanism of anthrax vaccine and provide the theoretical basis for vaccine development and evaluation, In this study, two kinds of Bacillus anthracis vaccine, Bacillus anthracis A16R vaccine and adsorbed PA vaccine, were used to observe the immune response mechanism of the two vaccines from three levels: spleen, bone marrow and peripheral blood, respectively. This study was divided into three parts: the preparation of two anthrax vaccines and the determination of their immune effects. Preparation and Identification of two anthrax Vaccines: dynamic observation of Serum Antibody titer and Antibody subtype Analysis of Balb/C mice immunized with A16R Bacillus Vaccine and Adsorbed PA Vaccine; dynamic observation of specific ASCs in Balb/C mice induced by A16R Bacillus Vaccine and Adsorbed PA Vaccine In this paper, we first established a ELISPOT method for the detection of antigen-specific antibody secreting cells in Balb/C mice immunized with anthrax vaccine. Bone marrow, spleen and peripheral blood were used to observe the dynamic changes and differences of specific ASCs in Balb/C mice immunized with two vaccines. The correlation between ASCs response and peripheral blood antibody titer in different organs / parts was analyzed. The spleen of Balb/C mice was induced by anthrax A16R spores and PA adsorbed vaccine. The dynamic changes of specific Th2 cell responses in bone marrow and peripheral blood, and the auxiliary effect of Th2 response in different organs / parts on the anti-spores / PA antibody in stable blood were analyzed in combination with the increase and decline of antibody titers in peripheral blood. Conclusion by observing the antibody titers in peripheral blood of Balb/C mice immunized with two anthrax vaccines, it was found that the antibody level of high titer could be detected in serum of mice with the enhanced immunization of the two vaccines and maintained for a long time. The level of IgG2a in the immunized group was lower than that in the control group, but IgG1 and IgG2b were still the main subtypes of the antibody subtype in the immunized group of the adsorbed PA vaccine, and the difference was significant compared with that of the control group, and the antibody subtypes of the immunized group were higher than that of the control group. ASCs were also detected in spleen, bone marrow and blood of Balb/C mice immunized with two kinds of vaccines, but the onset time and intensity of ASCs were different, the first appeared in spleen, and the peak value in bone marrow was later than that in spleen. However, the duration of response in bone marrow was longer, which was related to the maintenance of antibody titer in peripheral blood, and the dynamic changes of ASCs in spleen, bone marrow and peripheral blood of Balb/C mice after immunization with the two vaccines were different. Antigen-specific Th2 cell responses were detected in both bone marrow and peripheral blood, but the onset time and intensity of Th2 response were different, the first peak appeared in bone marrow, and the maintenance time was long. A strong Th2 response could be detected in peripheral blood. With the increase and maintenance of Th2 response in spleen, bone marrow and peripheral blood, the antibody titer of peripheral blood increased continuously, and remained at a high level 60 or 30 days after the last booster immunization. Vaccine immune response is a complex and multi-level process, which is regulated by complex network. Therefore, the evaluation of vaccine should be carried out from many aspects, angles, comprehensive, in-depth and systematic research.
【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】：2007
【分類號】：R392

【共引文獻(xiàn)】

相關(guān)期刊論文 前1條

1 陳茶;黃彬;姜儻;;一種細(xì)胞免疫學(xué)新技術(shù)——酶聯(lián)免疫斑點(diǎn)技術(shù)[J];國際檢驗(yàn)醫(yī)學(xué)雜志;2006年03期

相關(guān)博士學(xué)位論文 前3條

1 孫琳;HSP65-MUC1通過分子模擬損傷小鼠胰腺的可能性研究[D];吉林大學(xué);2007年

2 王永生;趨化并靶向到APCs的融合疫苗誘導(dǎo)抗腫瘤血管生成的細(xì)胞及體液免疫[D];四川大學(xué);2007年

3 李劍平;Notch信號通路在同基因造血干/祖細(xì)胞移植誘導(dǎo)免疫耐受中的作用[D];第四軍醫(yī)大學(xué);2008年

本文編號：1876341