本文選題：完整肽聚糖　切入點(diǎn)：重組乙肝表面抗原　出處：《重慶醫(yī)科大學(xué)》2005年碩士論文

【摘要】：雙歧桿菌完整肽聚糖有很強(qiáng)的免疫增強(qiáng)活性,但它在機(jī)體對(duì)弱免疫原重組乙肝表面抗原的免疫應(yīng)答中的作用尚不清楚。本課題旨在初步探討雙歧桿菌完整肽聚糖對(duì)重組乙肝表面抗原的佐劑效應(yīng)及其相關(guān)機(jī)制。 實(shí)驗(yàn)使用兩歧雙歧桿菌分離提純完整肽聚糖作為佐劑,采用 PBS 液混合、戊二醛或二磷酸腺苷交聯(lián)來吸附或結(jié)合重組乙肝表面抗原,并檢測其對(duì)抗原的吸附情況;制備免疫原,免疫 BALB/c 小鼠,觀察其對(duì)小鼠的毒副作用以及對(duì)胸腺和脾臟發(fā)育的影響;ELISA 法檢測小鼠血清的特異性抗體,LDH 釋放法檢測脾臟 NK細(xì)胞和 CTL 細(xì)胞體外殺傷活性,MTT 法檢測脾臟淋巴細(xì)胞體外增殖能力,ELISA法檢測脾臟淋巴細(xì)胞體外誘生的細(xì)胞因子。 結(jié)果顯示,雙歧桿菌完整肽聚糖以 PBS 液混合、戊二醛交聯(lián)或二磷酸腺苷交聯(lián)等方式均能吸附重組乙肝表面抗原結(jié)果,蛋白質(zhì)交聯(lián)劑的促進(jìn)作用并不明顯;免疫 BALB/c 小鼠后,未見毒副反應(yīng)出現(xiàn),生長情況正常,體重增長平穩(wěn),胸腺和脾臟明顯增重;免疫 8 周后,小鼠血清抗-HBs 陽性率提高和抗體滴度均比鋁佐劑提高,抗體中 IgG2a 亞型的滴度也明顯上升;脾臟淋巴細(xì)胞體外增殖能力增強(qiáng),體外誘生細(xì)胞因子的能力得到改善,NK 細(xì)胞和抗原特異性 CTL 細(xì)胞殺傷活性提高。 實(shí)驗(yàn)表明,雙歧桿菌完整肽聚糖在某些緩沖鹽溶液中可以吸附一定量的重組乙肝表面抗原;作為佐劑免疫 BALB/c 小鼠后,不出現(xiàn)毒副作用。雙歧桿菌完整肽聚糖能夠通過緩解胸腺衰老和增加脾重以維持淋巴器官功能,并通過增強(qiáng)脾臟淋巴細(xì)胞體外增殖能力和誘生細(xì)胞因子能力調(diào)節(jié)機(jī)體免疫應(yīng)答中 Th1/Th2 的平衡,增強(qiáng) NK 細(xì)胞和 CTL 細(xì)胞殺傷活性以增強(qiáng)機(jī)體對(duì)病毒感染細(xì)胞的清除能力,在獲得足夠保護(hù)性抗體的同時(shí)產(chǎn)生適當(dāng)細(xì)胞免疫應(yīng)答,展現(xiàn)出良好的免疫佐劑品質(zhì)。
[Abstract]:Bifidobacterium peptidoglycan has a strong immune enhancement activity, but its role in the immune response to weak immunogen recombinant hepatitis B surface antigen is unclear.The aim of this study was to investigate the adjuvant effect of complete peptidoglycan of Bifidobacterium on recombinant hepatitis B surface antigen and its related mechanism.The whole peptidoglycan was isolated and purified by bifidobacterium bifidobacterium as adjuvant. The recombinant hepatitis B surface antigen was adsorbed or combined with glutaraldehyde or adenosine diphosphate crosslinking with PBS solution.BALB/c mice were immunized,To observe its toxicity and side effects on the development of thymus and spleen in mice; to detect the activity of splenic NK cells and CTL cells in vitro by Elisa and to detect the lymphatic fineness of spleen by using Elisa method to detect the release of specific antibody against LDH and to detect the cytotoxicity of splenic NK cells and CTL cells in vitro.In vitro proliferative ability and Elisa were used to detect cytokines induced by splenic lymphocytes in vitro.The results showed that the complete peptidoglycan of Bifidobacterium could adsorb recombinant hepatitis B surface antigen by PBS solution, glutaraldehyde crosslinking or adenosine diphosphate crosslinking, but the effect of protein crosslinking agent was not obvious.There were no side effects, normal growth, steady weight gain, significant weight gain in thymus and spleen, and increased anti-HBs positive rate and antibody titer in serum of mice after 8 weeks of immunization compared with aluminum adjuvant.The titer of IgG2a subtype in antibody also increased obviously, the proliferation ability of splenic lymphocytes in vitro was enhanced, the ability of inducing cytokines in vitro was improved, and the killing activity of NK cells and antigen-specific CTL cells was improved.The results showed that Bifidobacterium peptidoglycan adsorb a certain amount of recombinant hepatitis B surface antigen in some buffer salt solution, and no side effects occurred after immunizing BALB/c mice as adjuvant.Bifidobacterium peptidoglycan can maintain lymphoid organ function by relieving thymus aging and increasing spleen weight, and regulate the balance of Th1/Th2 in immune response by enhancing the proliferation and inducing cytokine ability of splenic lymphocytes in vitro.To enhance the killing activity of NK cells and CTL cells in order to enhance the ability of the body to clear the virus infected cells, and to produce appropriate cellular immune response while obtaining sufficient protective antibodies, showing a good quality of immune adjuvant.
【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2005
【分類號(hào)】：R392

【共引文獻(xiàn)】

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