Novel Constructs of Tuberculosis Gene Vaccine and Its Immune Effect on Mice

摘要

＜正＞ A novel tuberculosis（TB）gene vaccine containing mouse granulocyte macrophage-colony stimulating factor（mGM-CSF）and a TB antigen（Ag85A）was developed in this study.The genes encoding Ag85A and mGM-CSFwere amplified by PCR respectively from the Ag85A-containing pBSby5 and pC-mGM-CSF.The genes were thencloned into two different polylinker sites of plasmid pIRES,forming a novel TB gene vaccine construct pI85AGM.Following transfection of pI85AGM plasmid into 7721 cell line by LipofectamineTM,the expression of Ag85A andGM-CSF proteins was identified by Western blotting or RT-PCR.Then Balb/c mice were inoculated with therecombinant pI85AGM,pI85A,pIGM or plasmid alone,respectively.The activities of CTL,NK cells and theAg85A-stimulated proliferation of spleen cells were measured by MTT method.The serum antibody against Ag85Awas detected by ELISA.The results showed that the Ag85A and GM-CSF proteins could be expressed in 7721 cellline and the activity of CTLs and the proliferation of spleen cells were significantly increased in thepI85AGM-immunized mice,indicating that the pI85AGM-immunized mice could generate specific immuneresponses to Ag85A.This study might provide possibility for developing novel anti-TB gene vaccine.Cellular ＆Molecular Immunology.2005;2（1）:57-62.Cellular ＆ Molecular Immunology.2005;2（1）:57-62.