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• W2272502034 abstract "AACR Annual Meeting-- Apr 12-16, 2008; San Diego, CA4992 Objective: To determine whether antigen presenting cells in malignant ascites (aAPC) can elicit an anti-tumor immune response after ex vivio stimulation with Toll-like receptor agonists (TLR-a) in a mouse model of ovarian cancer. Methods: C57/b6 mice were injected with murine ovarian cancer cell lines, ID8-Vegf-β-defensin and ID8-E7. Both aAPC and T cells harvested from ascites were subjected to flow cytometric characterization using antibodies to CD3, CD4, CD8, FoxP3, CD11c, CD14, CD80, CD86,CD123, B7-H1, and MHC II. Adherent CD45+ cells from ascites were considered candidate aAPC and stimulated with TLR-a, including LPS, CpG, and Poly I:C along with a blocking antibody to the IL-10 receptor (IL-10Rab) or to MHC-I. After 48 hours, treated aAPC were incubated with T cells harvested from autologous ascites specimens, and IFN-γ production was assessed by ELISA. Peripheral T cells from a second group of mice vaccinated with E7/E8 DNA were used as controls. To study the therapeutic potential of these cells, healthy mice were injected subcutaneously with a cellular vaccine composed of stimulated aAPC. After two weeks, peripheral T cells from vaccinated mice were incubated with stimulated aAPC in vitro . IFN-γ production was measured by ELISA. Vaccinated mice were then injected with tumor to evaluate for survival differences after immunization. Results: Cells from fresh ascites expressed CD80, CD86, and high levels of MHC II. After 48 hours in culture, stimulated cells expressed more CD14, CD80, CD86, B7-H1, and CD11c, but less CD123, than unstimulated cells. Functional studies showed high levels of IFN-γ production by T cells after incubation with aAPC treated with LPS, CpG, and IL-10Rab. Autologous ascites-derived T cells consistently produced more IFN-γ than control T cells. Addition of an anti-MHC I antibody blocked this effect, suggesting that CD8 cells were primarily responsible for IFN-γ production. Finally, peripheral T cells derived from healthy mice vaccinated with stimulated aAPC produced IFN-γ following exposure to stimulated aAPC in vitro. No IFN-γ was produced by T cells from mice vaccinated with unstimulated aAPC. Mice vaccinated with treated aAPC were less likely to develop tumor. Conclusions: APC present in malignant ascites have engulfed tumor antigen in vivo , and can be stimulated to elicit a polyclonal anti-tumor cellular immune response in a mouse model. This immunity is transferable as a cellular vaccine and confers protection from tumor challenge. This finding has significant translational potential as a rapid protocol for immunotherapy for select patients with ovarian cancer." @default.
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• W2272502034 date "2008-05-01" @default.
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• W2272502034 title "Development of a polyvalent cellular vaccine against ovarian cancer using ascites-derived antigen presenting cells" @default.
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