沙门氏菌菌蜕佐剂增强结核分枝杆菌4种蛋白的免疫作用研究
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摘要
为探究肠炎沙门氏菌菌蜕(ghost)佐剂对结核分枝杆菌(MTB) 4种抗原的免疫增强作用,本研究以MTB中的Rv3802、Rv0394c(SEC)、Rv0199及E6c10(ESAT6与CFP10融合蛋白)蛋白为免疫原,应用小鼠模型评价沙门氏菌菌蜕对MTB 4种抗原的免疫佐剂效应。结果显示,蛋白加菌蜕佐剂组可以刺激小鼠产生持久高水平的抗体,并且菌蜕佐剂的免疫增强效果高于弗氏不完全佐剂;IgG1/IgG2a的比率增高,提示蛋白加菌蜕佐剂组小鼠趋向于Th2型免疫反应,同时可以诱导淋巴细胞分化为CD4~+T细胞亚群并产生IFN-γ及IL-4细胞因子;病理组织学显示以菌蜕佐剂配伍蛋白制备的亚单位疫苗具有良好的安全性。本研究为结核病的预防和新型疫苗的研发奠定了一定的基础。
In order to evaluate the immunological enhancement of Salmonella enteritidis ghost adjuvant aiding four important antigens of Mycobacterium tuberculosis(MTB), the immunological efficiency of the four MTB proteins of Rv3802c, Rv0394c(SEC), Rv0199 and E6c10(ESAT6 and CFP10 fusion protein) associated with the bacterial ghost(BG) adjuvants were evaluated in mice model. The study demonstrated the BG adjuvant was able to stimulate mice to secrete persistent and higher levels of antigen-specific antibodies than that of Freund's incomplete adjuvant. The increasing ratio of IgG1/IgG2a demonstrated that the ghost adjuvant with the four proteins induce an immune response bias for Th2 in challenged mice. Simultaneously, the BG adjuvant also helped lymphocytes differentiate into substantial CD4~+T cells and produced cytokines of IFN-γ and IL-4. In addition,the pathological examination suggested that the BG was a safe adjuvant. These findings not only provide new subunit vaccine candidates against TB, but also pave a way for new adjuvant creation in the area of anti-TB vaccines.
In order to evaluate the immunological enhancement of Salmonella enteritidis ghost adjuvant aiding four important antigens of Mycobacterium tuberculosis(MTB), the immunological efficiency of the four MTB proteins of Rv3802c, Rv0394c(SEC), Rv0199 and E6c10(ESAT6 and CFP10 fusion protein) associated with the bacterial ghost(BG) adjuvants were evaluated in mice model. The study demonstrated the BG adjuvant was able to stimulate mice to secrete persistent and higher levels of antigen-specific antibodies than that of Freund's incomplete adjuvant. The increasing ratio of IgG1/IgG2a demonstrated that the ghost adjuvant with the four proteins induce an immune response bias for Th2 in challenged mice. Simultaneously, the BG adjuvant also helped lymphocytes differentiate into substantial CD4~+T cells and produced cytokines of IFN-γ and IL-4. In addition,the pathological examination suggested that the BG was a safe adjuvant. These findings not only provide new subunit vaccine candidates against TB, but also pave a way for new adjuvant creation in the area of anti-TB vaccines.
引文
[1]Sali M,Dainese E,Morandi M,et al.Homologous prime boosting based on intranasal delivery of non-pathogenic invasive Escherichia coli expressing MPT64,decreases Mycobacterium tuberculosis dissemination[J].Vaccine,2014,32(3):4051-4058.
[2]Colditz G A,Brewer T F,Fineberg H V,et al.Efficacy of BCGvaccine in the prevention of tuberculosis:Meta-analysis of the published literature[J].J Am Med Assoc,1994,271(6):698-702.
[3]Girard M P,Fruth U,Kieny M P.A review of vaccine research and development:Tuberculosis[J].Vaccine,2005,23(17):5725-5731.
[4]Langermans J A,Doherty T M,Greenwald R,et al.Protection of macaques against Mycobacterium tuberculosis infection by a subunit vaccine based on a fusion protein of antigen 85B and ESAT-6[J].Vaccine,2005,23(5):2740-2750.
[5]Coffman R L,Sher A,Seder R A.Vaccine adjuvants:Putting innate immunity to work[J].Immunity,2010,33(6):492-503.
[6]Liu Si-guo,Gong Qiang,Kong Xian-gang,et al.A novel DNAvaccine for protective immunity against virulent Mycobacterium bovis in mice[J].Immunol Lett,2008,117(8):136-145.
[7]Parker S K,Barkley R M,Rino J G,et al.Mycobacterium tuberculosis Rv3802c encodes a phospholipase/thioesterase and is inhibited by the antimycobacterial agent tetrahydrolipstatin[J].PLo S One 2009,4(1):e4281.
[8]Derrick S C,Yabe I M,Morris S,et al.Immunogenicity and protective efficacy of novel Mycobacterium tuberculosis antigens[J].Vaccine,2013,31(12):4641-4646.
[9]Lubitz W,Witte A,Eko F O,et al.Extended recombinant bacterial ghost system[J].J Biotechnol,1999,73(2-3):261-273.
[10]Rosenblum M D,Way S S,Abbas A K.Regulatory T cell memory[J].Nat Rev Immunol,2016,16(2):90-101.
[2]Colditz G A,Brewer T F,Fineberg H V,et al.Efficacy of BCGvaccine in the prevention of tuberculosis:Meta-analysis of the published literature[J].J Am Med Assoc,1994,271(6):698-702.
[3]Girard M P,Fruth U,Kieny M P.A review of vaccine research and development:Tuberculosis[J].Vaccine,2005,23(17):5725-5731.
[4]Langermans J A,Doherty T M,Greenwald R,et al.Protection of macaques against Mycobacterium tuberculosis infection by a subunit vaccine based on a fusion protein of antigen 85B and ESAT-6[J].Vaccine,2005,23(5):2740-2750.
[5]Coffman R L,Sher A,Seder R A.Vaccine adjuvants:Putting innate immunity to work[J].Immunity,2010,33(6):492-503.
[6]Liu Si-guo,Gong Qiang,Kong Xian-gang,et al.A novel DNAvaccine for protective immunity against virulent Mycobacterium bovis in mice[J].Immunol Lett,2008,117(8):136-145.
[7]Parker S K,Barkley R M,Rino J G,et al.Mycobacterium tuberculosis Rv3802c encodes a phospholipase/thioesterase and is inhibited by the antimycobacterial agent tetrahydrolipstatin[J].PLo S One 2009,4(1):e4281.
[8]Derrick S C,Yabe I M,Morris S,et al.Immunogenicity and protective efficacy of novel Mycobacterium tuberculosis antigens[J].Vaccine,2013,31(12):4641-4646.
[9]Lubitz W,Witte A,Eko F O,et al.Extended recombinant bacterial ghost system[J].J Biotechnol,1999,73(2-3):261-273.
[10]Rosenblum M D,Way S S,Abbas A K.Regulatory T cell memory[J].Nat Rev Immunol,2016,16(2):90-101.