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

Abstract

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 adenoviruses,negative Ad-eGFP group which was adding with blank adenovirus 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 successfully 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 adenovirus 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-γ. Key words: Dendritic cells; Mouse alpha-fetoprotein; 4-1BBL gene; Biology characteristics