本文選題：治療性疫苗　切入點：減毒鼠傷寒沙門氏菌　出處：《復旦大學》2010年博士論文

【摘要】： 治療性疫苗能夠打破免疫耐受并引起針對癌癥細胞的長時程免疫反應,由于疫苗給藥途徑與構建有效的抗原遞呈系統(tǒng)是疫苗制備需要解決的關鍵性問題。減毒鼠傷寒沙門氏菌作為癌癥疫苗的載體能夠將DNA從原核細胞傳遞至真核細胞,并且能夠選擇性積聚在腫瘤組織。熱休克蛋白70能夠通過其多肽結合結構域與抗原結合并與抗原遞呈細胞表面的Hsp70受體作用,將抗原傳遞至抗原遞呈細胞。本論文的第一部分,我們構建了一種新型的低拷貝DNA疫苗,該疫苗基于Hsp70-TAA復合物構建,并由減毒鼠傷寒沙門氏菌SL3261菌株作為載體將DNA運送至小鼠體內�？诜撝亟M細菌疫苗能夠誘發(fā)針對黑色素腫瘤細胞的CTL細胞毒性細胞殺傷作用并顯著激活T細胞反應。在腫瘤預防實驗組C57BL/6J小鼠模型中,該治療性口服疫苗能夠誘發(fā)57.1%的腫瘤預防免疫反應,在腫瘤治療組中,該疫苗能夠完全消除62.5%的C57BL/6J小鼠中B16F10腫瘤的生長,顯著延長小鼠的生存期。 口服減毒沙門氏菌疫苗載體通常定位在吞噬性細胞中,限制了其所攜帶的抗原蛋白進入抗原遞呈途徑,論文的第二部分我們利用沙門氏菌Ⅲ型分泌系統(tǒng)對蛋白質的轉運功能來克服這種限制。Ⅲ型分泌蛋白能夠將沙門氏菌編碼的抗原蛋白直接運送至抗原遞呈細胞胞質�？乖f呈細胞表面存在Hsp70的特異性受體,故Hsp70能夠以Hsp70-TAA復合物的形式向APC細胞遞呈抗原。本研究利用Ⅲ型分泌系統(tǒng)來實現(xiàn)對Hsp70-TRP2融合抗原蛋白的轉運和遞呈。口服該重組細菌疫苗能夠實現(xiàn)對腫瘤特異性抗原的有效遞呈,打破免疫耐受,誘導產(chǎn)生腫瘤細胞特異性的CTL反應和NK細胞殺傷作用,并在C57BL/6J小鼠上建立的B16F10黑色素腫瘤模型中獲得了75%的腫瘤預防效果和62.5%的治療效果。我們設計了以減毒鼠傷寒沙門氏菌作為腫瘤DNA疫苗及蛋白疫苗的傳遞工具的兩套抗原遞呈系統(tǒng),為腫瘤的預防和治療提供了有效的新方法。
[Abstract]:Therapeutic vaccine can break the immune tolerance and induce long-term immune response to cancer cells. Because of the vaccine delivery pathway and the construction of an effective antigen presentation system is the key problem to be solved in vaccine preparation.Attenuated Salmonella typhimurium can transfer DNA from prokaryotic cells to eukaryotic cells as a vector of cancer vaccine and can selectively accumulate in tumor tissues.Heat shock protein 70 (HSP70) can transfer antigens to antigen-presenting cells through its polypeptide binding domain which binds to antigens and acts with Hsp70 receptors on the surface of antigen-presenting cells.In the first part of this thesis, we constructed a novel low copy DNA vaccine, which was constructed based on Hsp70-TAA complex and was transported to mice by attenuated Salmonella typhimurium SL3261 strain as a vector.Oral administration of the recombinant bacterial vaccine could induce cytotoxicity of CTL cells against melanoma cells and activate T cell response significantly.In the C57BL/6J mice model of tumor prevention group, the therapeutic oral vaccine could induce 57.1% tumor preventive immune response. In the tumor treatment group, the vaccine could completely eliminate the growth of B16F10 tumor in 62.5% of C57BL/6J mice.The survival time of mice was significantly prolonged.Oral attenuated Salmonella vaccine vectors are usually located in phagocytic cells, limiting the entry of antigen proteins into antigen presentation.In the second part of this paper, we use the protein transport function of Salmonella type 鈪，

本文編號：1725736