lnterleukin-21(IL-21)is a recently characterized T cell-derived cytokine with a significant homology to IL-2, IL-4 and IL-15.To determine whether IL-21 has broad immunoregulatory activity and can stimulate durable ant...lnterleukin-21(IL-21)is a recently characterized T cell-derived cytokine with a significant homology to IL-2, IL-4 and IL-15.To determine whether IL-21 has broad immunoregulatory activity and can stimulate durable antitumour responses,we constructed mouse IL-21(mIL-21) recombinant plasmid and evaluated its antitumor efficacy.Mouse IL-21 cDNA was amplified from Con A-activated mouse T cells by RT-PCR.Recombinant pcDNA3.1/mIL-21 was constructed and transfected into Sp2/0 cells.Mouse IL-21 expression was analyzed by Western blotting and its activities were detected by ~3H-TdR incorporation and MTT assay.The recombinant pcDNA3.1/mIL-21 was injected s.c.into tumor lump.Tumor size,weight,the activities of CTLs,NK cells and LAK cells and serum IFN-γ,level were measured for evaluating mIL-21 mediated antitumor responses.The results indicated that mIL-21 was correctly expressed in Sp2/0 cells and it can improve the proliferation of T cells and B cells,and enhance NK cytotoxic activity in vitro.The activities of CTL and NK cells,and serum IFN-γlevel were significantly improved,furthermore the tumor growth was obviously suppressed in pcDNA3.1/mIL-21 treated mice.However,the LAK activity did not alter significantly.Taken together,this study suggests that the injection with recombinant plasmid containing mIL-21 is a potential approach for tumor gene therapy.Cellular & Molecular Immunology.2004;1(6):461-466.展开更多
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-CSF were amplified by...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-CSF were amplified by PCR respectively from the Ag85A-containing pBSby5 and pC-mGM-CSF.The genes were then cloned 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 Lipofectamine^(TM),the expression of Ag85A and GM-CSF proteins was identified by Western blotting or RT-PCR.Then Balb/c mice were inoculated with the recombinant pI85AGM,pI85A,pIGM or plasmid alone,respectively.The activities of CTL,NK cells and the Ag85A-stimulated proliferation of spleen cells were measured by MTT method.The serum antibody against Ag85A was detected by ELISA.The results showed that the Ag85A and GM-CSF proteins could be expressed in 7721 cell line and the activity of CTLs and the proliferation of spleen cells were significantly increased in the pI85AGM-immunized mice,indicating that the pI85AGM-immunized mice could generate specific immune responses 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.展开更多
文摘lnterleukin-21(IL-21)is a recently characterized T cell-derived cytokine with a significant homology to IL-2, IL-4 and IL-15.To determine whether IL-21 has broad immunoregulatory activity and can stimulate durable antitumour responses,we constructed mouse IL-21(mIL-21) recombinant plasmid and evaluated its antitumor efficacy.Mouse IL-21 cDNA was amplified from Con A-activated mouse T cells by RT-PCR.Recombinant pcDNA3.1/mIL-21 was constructed and transfected into Sp2/0 cells.Mouse IL-21 expression was analyzed by Western blotting and its activities were detected by ~3H-TdR incorporation and MTT assay.The recombinant pcDNA3.1/mIL-21 was injected s.c.into tumor lump.Tumor size,weight,the activities of CTLs,NK cells and LAK cells and serum IFN-γ,level were measured for evaluating mIL-21 mediated antitumor responses.The results indicated that mIL-21 was correctly expressed in Sp2/0 cells and it can improve the proliferation of T cells and B cells,and enhance NK cytotoxic activity in vitro.The activities of CTL and NK cells,and serum IFN-γlevel were significantly improved,furthermore the tumor growth was obviously suppressed in pcDNA3.1/mIL-21 treated mice.However,the LAK activity did not alter significantly.Taken together,this study suggests that the injection with recombinant plasmid containing mIL-21 is a potential approach for tumor gene therapy.Cellular & Molecular Immunology.2004;1(6):461-466.
文摘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-CSF were amplified by PCR respectively from the Ag85A-containing pBSby5 and pC-mGM-CSF.The genes were then cloned 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 Lipofectamine^(TM),the expression of Ag85A and GM-CSF proteins was identified by Western blotting or RT-PCR.Then Balb/c mice were inoculated with the recombinant pI85AGM,pI85A,pIGM or plasmid alone,respectively.The activities of CTL,NK cells and the Ag85A-stimulated proliferation of spleen cells were measured by MTT method.The serum antibody against Ag85A was detected by ELISA.The results showed that the Ag85A and GM-CSF proteins could be expressed in 7721 cell line and the activity of CTLs and the proliferation of spleen cells were significantly increased in the pI85AGM-immunized mice,indicating that the pI85AGM-immunized mice could generate specific immune responses 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.
