The infection of dendritic cells by recombinant adenoviral vector carrying HBsAg-HSP70 chimeric gene and its biological characteristics observations

• Published in: Zhonghua shi yan he lin chuang bing du xue za ji Vol. 23; no. 1; p. 29
• Main Authors: Lei, Chun-Liang, Ou, Yang-Ling, Yang, Zhan, Tang, Xiao-Ping
• Format: Journal Article
• Language: Chinese
• Published: China 01-02-2009
• Subjects: Adenoviridae - genetics; Adenoviridae - physiology; Cells, Cultured; Cytokines - genetics; Cytokines - immunology; Dendritic Cells - immunology; Dendritic Cells - virology; Genetic Vectors - genetics; Hepatitis B Surface Antigens - genetics; Hepatitis B Surface Antigens - immunology; HSP70 Heat-Shock Proteins - genetics; HSP70 Heat-Shock Proteins - immunology; Humans; Lymphocyte Culture Test, Mixed; Recombinant Fusion Proteins - genetics; Recombinant Fusion Proteins - immunology
• ISSN: 1003-9279
• DOI: 10.3760/cma.j.issn.1003-9279.2009.01.011

To test the infeciton efficiency of recombinant adenoviral vector carrying HBsAg-HSP70 chimeric gene and to abserve its biological characteristics.
Peripheral blood mononuclear cells (PBMC) were separated from healthy blood donor and they were infected by Ad-HSP70-HBsAg on the first day of isolation. DCs were induced in medium with cytokines IL-4, GM-CSF and TNF-alpha in vitro. The biological characteristics of DC induced were analyzed by inverted fluorecent microscope, RT-PCR, flow cytometer (FACS), and mixed lymphocyte reaction (MLR).
The traced gene-GFP were abserved in DCs by inverted fluorecent microscope and HSP70-HBsAg gene mRNA expression was detected by RT-PCR after the Ad-HSP70-HBsAg infection. FACS analysis shown that the expression of CD1a, CD80, CD86 and HLA-DR on surfece of two groups of DCs were similar. MLR showed that there are not a statitic difference of stimulated index (SI) between two groups.
Results indicated that Ad-HSP70-HBsAg can effectively infected DCs without affecting its biological characteristics.