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Objective: To investigate whether human dendritic cells (DC) derived from peripheral blood mononuclear cells (PBMC), which were pulsed by heat shock protein 70 (HSP70) isolated from human bladder tumor cell lines of EJ, were able to induce peptide specific cytotoxic T-lymphocytes (CTL) response in vitro and give the experimental foundation for the future clinical trials of immunotherapy in bladder tumor. Methods: The EJ-derived HSP70 co-cultured with DC from the healthy volunteers’ PBMC, along with the crude lysate (the supernatant before HSP70 purification) from EJ cells were used as the experimental groups and DC not pulsed by any tumor cells antigen were the blank control. The autologous T-lymphocytes were added into the above various DC groups, and after incubation, the stimulation indexes (SI) and interferon-γ (IFN-γ) were detected to evaluate the immune activities of various DC groups. The killing effects of CTL to target cells, EJ and Hela cells, were determined with 51Cr releasing test. Results: Both DC/HSP70 and DC/the crude lysate could effectively activate CTL in vitro and kill target cells EJ. The killing effect of DC/HSP70 to EJ was much stronger than DC/the crude lysate (the supernatant before HSP70 purification) (P < 0.05). DC without any tumor cell antigens had a lower killing power to EJ. Meanwhile, DC/ HSP70 had little killing power to Hela non-relevant to bladder tumor histopathologically as compared with EJ cells (P < 0.05). Conclusion: The DC pulsed by HSP70 derived from the autologous tumor cells could induce a peptide complexes specific CTL response to tumor cells, and the CTL response induced by the DC/HSP70 was stronger, which display the basis of the possible clinical application of DC/HSP70 for bladder tumor.
Objective: To investigate whether human dendritic cells (derived from peripheral blood mononuclear cells (PBMC), which were pulsed by heat shock protein 70 (HSP70) from human bladder tumor cell lines of EJ, were able to induce peptide specific cytotoxic T Methods: The EJ-derived HSP70 co-cultured with DC from the healthy volunteers’ PBMC, along with the crude lysate (the supernatant before HSP70 purification) from EJ cells were used as the experimental groups and DC not pulsed by any tumor cells antigen were the blank control. The autologous T-lymphocytes were added into the above various DC groups, and after incubation, the stimulation indexes ( SI) and interferon-γ (IFN-γ) were detected to evaluate the immune activities of various DC groups. The killing effects of CTL to target cells, EJ and Hela cells, were determined with 51Cr rele asing test. Both DC / HSP70 and DC / the crude lysate could effectively activate CTL in vitro and kill target cells EJ. The killing effect of DC / HSP70 to EJ was much stronger than DC / the crude lysate (the supernatant before HSP70 DC without any cell killing antigens had a lower killing power to EJ. However, DC / HSP70 had little killing power to Hela non-relevant to bladder tumor histopathologically as compared with EJ cells (P <0.05) . Conclusion: The DC pulsed by HSP70 derived from the autologous tumor cells could induce a peptide complexes specific CTL response to tumor cells, and the CTL response induced by the DC / HSP70 was stronger, which display the basis of the possible clinical application of DC / HSP70 for bladder tumor.