本文選題：B族鏈球菌　切入點(diǎn)：多糖蛋白質(zhì)結(jié)合物　出處：《蘭州大學(xué)》2010年碩士論文　論文類型：學(xué)位論文

【摘要】： 目的應(yīng)用B族鏈球菌莢膜多糖-破傷風(fēng)類毒素結(jié)合物(GBS-CPS-TT)及B族鏈球菌莢膜多糖-白喉類毒素結(jié)合物(GBS-CPS-DT),經(jīng)不同免疫途徑免疫小鼠,分別測定小鼠血清中特異性IgG的抗體滴度,觀察并比較兩種不同載體的B族鏈球菌莢膜多糖-蛋白質(zhì)結(jié)合物經(jīng)黏膜途徑誘導(dǎo)的全身特異性免疫反應(yīng)狀況,探索最佳黏膜免疫途徑及免疫劑量。 方法選取體重14-16g健康清潔級(jí)NIH品系雌性小鼠300只,隨機(jī)取240只分別以GBS-CPS-TT、GBS-CPS-DT免疫,分為皮下注射5μg組、滴鼻5μg組、滴鼻10μg組和灌胃10μg組共8組,每組各30只。各組均免疫3次,每次免疫間隔時(shí)間2周,免疫后1周時(shí)采血,分離血清,低溫保存。采用間接酶聯(lián)免疫實(shí)驗(yàn)(ELISA)檢測小鼠血清中特異性IgG的抗體滴度。 結(jié)果GBS-CPS-TT、GBS-CPS-DT結(jié)合物滴鼻免疫、灌胃免疫和皮下注射均可以誘導(dǎo)產(chǎn)生特異性血清IgG抗體。滴鼻10μg組誘導(dǎo)產(chǎn)生的IgG抗體滴度明顯高于滴鼻5μg組(P0.01)；當(dāng)免疫劑量相同時(shí),滴鼻免疫誘導(dǎo)產(chǎn)生的IgG抗體滴度顯著低于皮下注射產(chǎn)生的IgG抗體滴度(P0.01)。作為載體蛋白TT與DT比較無差異(P0.05)。對(duì)照組未檢出特異性IgG抗體。 結(jié)論GBS-CPS-TT、GBS-CPS-TT結(jié)合物經(jīng)滴鼻免疫小鼠后可產(chǎn)生明顯的系統(tǒng)性免疫應(yīng)答,滴鼻10μg組的免疫效果優(yōu)于5μg組,抗體滴度有顯著性差異意義,表明較大的免疫劑量達(dá)到的免疫效果可能更優(yōu)；黏膜免疫后第二、三針誘導(dǎo)應(yīng)答產(chǎn)生的抗體滴度具有明顯的遞增增強(qiáng)效應(yīng),表明結(jié)合物有免疫記憶及加強(qiáng)免疫應(yīng)答效應(yīng)；GBS多糖蛋白質(zhì)結(jié)合物的黏膜免疫機(jī)制還有待進(jìn)一步研究。
[Abstract]:Objective to immunize mice with group B streptococcus perfringens polysaccharide tetanus toxoid conjugate (GBS-CPS-TTT) and group B streptococcus perfringens polysaccharide diphtheria toxoid conjugate (GBS-CPS-DTT), and determine the antibody titers of specific IgG in serum of mice. To observe and compare the systemic specific immune response induced by mucosal pathway of two different carriers of streptococcus B group capsule polysaccharides and protein conjugates, and to explore the best mucosal immune pathway and immune dose. Methods A total of 300 healthy and clean grade NIH strains of female mice weighing 14-16 g were selected and immunized with GBS-CPS-CPS-DT. 240 mice were divided into 5 渭 g subcutaneous injection group, 5 渭 g nasal drip group, 10 渭 g dripping nose group and 10 渭 g gavage group with 30 rats in each group. The blood samples were collected at 1 week after immunization, the serum was isolated and preserved at low temperature. The antibody titers of specific IgG in serum of mice were detected by indirect enzyme-linked immunosorbent assay (Elisa). Results the specific serum IgG antibody was induced by nasal immunization with GBS-CPS-TT conjugate GBS-CPS-DT, intragastric immunization and subcutaneous injection. The titer of IgG antibody in 10 渭 g group was significantly higher than that in 5 渭 g group, and the titer of IgG antibody was significantly higher in 10 渭 g group than that in 5 渭 g group. The titer of IgG antibody induced by nasal immunization was significantly lower than that produced by subcutaneous injection of IgG antibody (P 0.01). There was no difference between TT and DT as carrier protein (P 0.05). No specific IgG antibody was detected in the control group. Conclusion GBS-CPS-TTS-TT conjugate can produce obvious systemic immune response after nasal drip immunization in mice. The immune effect of 10 渭 g group is better than that of 5 渭 g group, and there is significant difference in antibody titer, which indicates that the immune effect reached by larger immune dose may be better than that in 5 渭 g group. The titer of antibody induced by three needles was significantly increased after mucosal immunization, which indicated that the conjugate had immune memory and enhanced immune response. The mucosal immune mechanism of GBS polysaccharide protein conjugate was still to be further studied.
【學(xué)位授予單位】：蘭州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2010
【分類號(hào)】：R392

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