Dendritic cells modified with 6Ckine/IFNγ fusion gene induce specific cytotoxic T lymphocytes in vitro
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Backgroud and objective
Dendritic cells play an important role in initiation and regulation of immune responses. Previous studies demonstrated that intratumoral administration of 6Ckine-modified DCs enhanced local and systemic antitumor effects. Herein we report the investigation of the specific CTL responses elicited by adenoviral 6Ckine/IFNγ fusion gene-modified DCs in vitro.
Human monocyte-derived DCs were modified with an adenoviral vector encoding 6Ckine/IFNγ fusion protein (Ad-6Ckine/IFNγ), and then investigated the effect of 6Ckine/IFNγ fusion protein on the maturation, cytokine and chemokine secretion of DCs, and their activities of recruiting and activating T cells in vitro were investigated.
6Ckine/IFNγ fusion protein induced DC maturation characterized with the upregulation of CD83 and CCR7. And it up-regulated the expression of RANTES and IL-12p70, down-regulated that of IL-10 in DCs. Additionally, 6Ckine/IFNγ markedly increased DC’s recruiting ability for naive T cells, benefiting from the enhanced expression of chemokines 6Ckine and RANTES in DCs. Fusion gene-modified DCs significantly promoted the proliferation of autologous T cells, induced Th1 differentiation by upregulating the expression of IL-2 and T-bet in T cells, and increased specific cytotoxicity of CTLs against specific tumor cells, HepG2 or LoVo cells, respectively.
Combining the effects of 6Ckine and IFNγ, Ad-6Ckine/IFNγ modified DCs induced enhanced CTL responses in vitro, which indicated that Ad-6Ckine/IFNγ modified DCs might be used as an adjuvant to trigger an effective antitumor immune response.
KeywordsDendritic cells Cytokines Chemokines Tumor immunology Gene therapy
Recombinant adenovirus carrying 6Ckine/IFNγ fusion gene
Recombinant adenoviruses carrying 6Ckine gene
Recombinant adenoviruses carrying IFNγ gene
Recombinant adenoviruses carrying β-galactosidase gene
Recombinant adenoviruses carrying green fluorescent protein gene
DC transfected with Ad-6Ckine
DC transfected with Ad-6Ckine/IFNγ
DC transfected with Ad-IFNγ
DC transfected with Ad-LacZ
National Institute for the Control of Pharmaceutical and Biological Products
This study was supported by the grants from National Basic Research Program of China (No.2004CB518801), Research & Development Fund of Guangdong Provine (No.2003A10902), Natural Science Foundation of Guangdong Province (No.021810), Research & Development Fund of Guangzhou City (No.2004Z3-E4011), and Medical Science Grant from Guangdong Province (No.B2002050). We thank Miao-la Ke, Jie-min Chen, Ying-hui Zhu and Xia Xiao (Sun Yat-sen University) for their technical assistance. We thank Dr. Changyou Wu, Dr. Li-min Zeng, and Dr. Qiang Liu (Sun Yat-sen University) for their review of this article. We also thank Ying-jun Ji (Doublle Bioproduct Inc.) and Yin Duan (Simon Fraser University) for their linguistic help in the preparation of this manuscript.
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