摘要
Objective： Heat shock protein （HSP） has the promiscuous abilities to chaperone and present a broad repertoire of tumor antigens to antigen presenting cells including DCs. In this report, we analyzed the modulation of immature DC by lISP 96 （gp96）. Method： Murine bone marro6-derived DC was induced by GM-CSF plus IL-4, which aped the immunostimulatory effects of DC. Cocultured DC and gp96-peptide complexes （gp96-PC） or inactivated H22 cells, the expression of MHC class Ⅱ , CD40, CD80 was quantified by flow cytometry. The concentration of IL-12 and TNF- in culture supematants were determined by ELISA. C1 release assay was used to test specific cytotoxic T cell. Results： Our study demonstrated that the extent of DC maturation induced by gp96-PC, which was reflected in surface density of costimulatory and MHC Ⅱmolecules, was correlated with the secretion of IL-12 and with the T cellactivating potential in vitro. Conclusion： Heat shock protein 96 could be isolated and purified from H22 cells and could induce maturation of dendritic cell. Our findings might be relevance to the use of DC vaccine in therapy of human tumors.
Objective： Heat shock protein （HSP） has the promiscuous abilities to chaperone and present a broad repertoire of tumor antigens to antigen presenting cells including DCs. In this report, we analyzed the modulation of immature DC by lISP 96 （gp96）. Method： Murine bone marro6-derived DC was induced by GM-CSF plus IL-4, which aped the immunostimulatory effects of DC. Cocultured DC and gp96-peptide complexes （gp96-PC） or inactivated H22 cells, the expression of MHC class Ⅱ , CD40, CD80 was quantified by flow cytometry. The concentration of IL-12 and TNF- in culture supematants were determined by ELISA. C1 release assay was used to test specific cytotoxic T cell. Results： Our study demonstrated that the extent of DC maturation induced by gp96-PC, which was reflected in surface density of costimulatory and MHC Ⅱmolecules, was correlated with the secretion of IL-12 and with the T cellactivating potential in vitro. Conclusion： Heat shock protein 96 could be isolated and purified from H22 cells and could induce maturation of dendritic cell. Our findings might be relevance to the use of DC vaccine in therapy of human tumors.
基金
National Natural Science Foundation of China (No.30200369)
