| Ovarian carcinoma remains the most lethal of gynecologic malignancies. Cytoreductive surgery followed by platinum-based combination chemotherapy is the standard treatment of patients with ovarian carcinoma. However, the overall 5-year survival has changed little over the last two decades. A new approach for the treatment is necessary to improve the prognosis.Tumor immune response requires effective antigen presentation. Dendritic cells are most potent antigen presenting cells known up to date. Highly- expressing MHC-I, II and co-stimulatory molecules, DC play a core role in tumor antigen presentation. In view of the effects of unmethylated CpG motif that containing oligonucleotides (CpG ODN),the anti-tumor immunity of Dendritic cells (DC) have not yet been studied, we investigate the influence of CpG ODN-2006 on human being cord blood derived dendritic cells (CDC) and and on the anti-tumor ability of Cytotoxic T lymphocytes (CTL)in vitro. ON the bases of the before studies, We also investigate whether CpG ODN can affect the antitumor responses of DC tumor cell vaccine against Ovarian carcinoma .We study by following methods: NO 1 CpG oligonucleotide2006 (ODN 2006) were used to promote maturation of DC in vitro.â‘ DC were induced from human being cord bloods by granulocyte-macrophage colony stimulating factor (GM-CSF), interleukin-4 (IL-4). CpG ODN2006 were added stimulating groups. DC were separated by a method of density gradient centrifugation of freshly isolated procedure with lymphocytes separation medium the mononuclear cells (MNCs) from cord blood.â‘¡DC were stimulated by CpG ODN2006 or not by different time and different concentrations. The expression ofCD86,HLA-DR,CD1a\ MHC-I on DC surface was analyzed by flow cytometer (FCM).â‘¢The level of interleukin-12 (IL-12) p35,p40 on cells membrance was detected by RT-PCR.â‘£The proliferation of T cells that was stimulated by CpG ODN activating DC was examined by MTT assay. NO 2 By fusing DC with ovarian carcinoma HO8910 cells (HO8910/DC) or culturing with TAA-tumor associating antigen (TAA/DC), We get stimulated groups and non-stimulated DC vaccine by CpG ODN2006.Fused HO8910 cells to DC were stained with PKH26 red fluorescence, and the fusion cells (HO8910/DC) were selected by flow cytometer ,then DC based tumor cell vaccines were developed. To determine the immune responses to the vaccines, T cell proliferation and cytotoxicity were done in vitro.â‘ We proceed T cell proliferation and cytotoxicity in vitro by using CpG ODN2006 stimulating DC vaccines and non ODN2006 stimulating DC vaccines respectively.â‘¡The expressions of CD86,HLA-DR,CD1a,MHC-I of different tumor antigen loading styles on DC surface were analyzed by flow cytometer (FCM). NO3 Therapeutic and prophylactic immunization with different DC vaccines were performed in BALB/ C mice bearing Ovarian carcinoma.We get results by the studies:â‘ Cord blood cells cultured in the presence of GM-CSF and IL-4 plus additional CpG ODN2006 appeared typical morphology of DC. FACS analyses showed that the mean fluorescence index (MFI) of CD1a,CD86,HLA-DR,MHC-I expression of DC stimulated with CpG ODN2006 were higher than that of without CpG ODN2006.â‘¡IL-12 from the DC cultured with CpG ODN2006 was higher than that from the DC cultured without CpG ODN2006.â‘¢T cell proliferation and T cell mediated cytotoxicity against HO8910 by MTT were induced in vitro. Marked inhibition of tumor growth in HO8910 bearing mice was observed upon prophylactic and therapeutic immunizations with the vaccines.We get following Conclusion: CpG ODN2006 can enhance the antitumor responses of DC vaccine and effectively produce differential antitumor immunity by promoting DC maturation. Immature DC engender definite antitumor ability by different antigen loading styles. And that mature DC get effective antitumor activity by the means of fusion cells to loading antigen.
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