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Large-Scale Expansion of Leukemia Specific CTLs Using Lysates-Pulsed Dendritic Cell and Low-Dose IL-2 for Adoptive Immunotherapy
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Abstract:
Objective: To determine the parameters settings for in vivo induction and in vitro expansion of leukemia-specific CTL. Methods: C57BL/6 mice were vaccinated repeatedly with 1×10 7 DCs pulsed with 5×10 7 FBL3-lysates (FBL3-LY) every 10 days. mice were sacrificed and the spleen T cells were collected and purified. Five days after the third immunization, T cells were restimulated weekly with FBL3-HS pulsed DC(30 Gy irradiated), and low-dose IL-2 was added 10 days following. Results: Three weeks′ co-culture of splenocytes T cells from the vaccinated mice with FBL3-LY pulsed DC in the presence of low dose IL-2 resulted in more than 100-folds′ expansion of leukemia specific CTLs, which had a higher percentage of CD8 + T cell. The expanded CTLs had potent cytotoxic activity against the parental FBL3 cells（81.84±8.68%） tested in standard LDH release assay, but no cytolytic activity above background was observed against the k562 leukemia targets. Conclusion: This regime could be considered as practical alternatives to the existing clinical immunotherapy strategies.
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