小鼠甲胎蛋白和4-1BB配体融合基因疫苗对树突状细胞生物学特性的影响  

The affects on the biology characteristics of dendritic cells transduced with truncated mouse alpha- fetoprotein internal ribosomal entry site 4-1BBL recombinant adenoviruses

作　　者：江志清[1] 张孝斌[2] 

机构地区：[1]武汉大学人民医院普外科,430060 [2]武汉大学人民医院泌尿外科,430060

出　　处：《中华实验外科杂志》2012年第10期1927-1929,共3页Chinese Journal of Experimental Surgery

摘　　要：目的观察感染小鼠甲胎蛋白（mAFP）和4-1BB配体（m4—1BBL）融合基因对小鼠树突状细胞（DC）生物学特性的影响。方法构建含mAFP和m4—1BBL的融合基因腺病毒Ad-mAFP-内部核糖体起始位点（IRES）-m4-1BBL，感染H22肝癌细胞，Westernblot法检测mAFP和4—1BBL蛋白的表达；诱导培养DC，并将其分为实验组、载体对照组和阴性对照组，以上各组感染后观察DC形态学变化；流式细胞仪检测各组细胞表型；混合淋巴细胞反应（MLR）检测各组DC的刺激活性以及对T细胞分泌干扰素（IFN）-1的影响。结果成功构建融合基因重组腺病毒Ad—mAFP-IRES-m4-1BBL并表达于H22细胞；体外诱导培养获得典型的DC；流式细胞仪检测各组DC感染后的细胞表型，实验组DC细胞CD80[（60．7±8．7）％]、CD86[（67．0±4．6）％]表达率显著增高于其余两组（P〈0．05），刺激T细胞增殖能力（0．66±0．05）亦显著增强（P〈0．05），刺激T细胞分泌IFN-γ[（1280．7±68．2）ng／L]水平更高（P〈0．05）。结论成功构建含mAFP及4．1BBL融合基因腺病毒疫苗，其感染DC后刺激T细胞增殖和分泌IFN-γ的能力增强。Objective To investigate the biological characteristics of the dendritic cells transduced with truncated mouse alpha-fetoprotein （mAFP） and 4-1BBL recombinant adenoviruses. Methods Construction with mAFP and m4-1BBL coexpression gene recombinant adenovirus, H22 cells infected by recombinant adenovirus, and then mAFP and 4-1BBL expression detected by Western blotting. C57BL/6j murine bone marrow derived dendritic cells inducted. The dendritic cells （DCs） will be divided into experimental group which was adding with recombinant adenovirnses, negative Ad-eGFP group which was adding with blank aderiovirus vectors and negative control group which was adding with normal saline, and then the morphological changes of DCs were observed by phase contrast microscope, flow cytometry was used to detective the phenotype of DCs, mixed lymphocyte reaction （MLR） was used to observe the DCs stimulating activity on T cell proliferation and secretion of cytokines. Results Recombinant adenovirus was success- fully constructed and can be expressed in H22 cells. Through the induction and culture in vitro, the mature DCs with typical figure can be found under the microscope, the DCs cell phenotype CD80, CD86 has significant difference between the experimental group and the other two groups （P 〈 0. 05）, the proliferation ability of DCs stimulated T cell was enhanced in the experimental group （P 〈 0. 05 ）, the DCs stimulated T cells secrete more interferon （IFN） -γ cytokines （ P 〈 0. 05 ）. Conclusion The recombinant adenovirns vaccine with fusion gene of mAFP and 4-1BBL was successfully constructed which transfected can increase the DC capacity to stimulate T cell proliferation and secretion of IFN-3.

关 键 词：树突状细胞 小鼠甲胎蛋白 4-1BB配体 生物学特性 

分 类 号：R392.1[医药卫生—免疫学]