Equal numbers of the two populations were mixed and injected challenged with 1 ! 106 live EG-7 cells and the survival and i.v. into mice. 18–48 h later, splenocytes from the recipients were tumor volume were monitored. Tumor volume based on caliper analyzed by flow cytometer to assess the peptide-specific killing. measurements were calculated by the modified ellipsoidal The ovalbumin-specific lysis was calculated as follows: r = % CFSElow / % CFSEhigh and % lysis = [1 – (runprimed / rprimed)] ! 100, where r is the ratio. formula, 1/2(Length ! Width2). Statistics. Statistical values were assessed by the Student’s t-test. P values were expressed and error bars are SE. The correlation of L. monocytegenes infection and tumor challenge. C57BL/6 tumor volume was analyzed for significance using 2-way analysis mice were vaccinated with the indicated cellular vaccine (day 0) of variance (ANOVA).The Kaplan-Meier method was used to before they were i.v. challenged with 5 ! 104 live L. monocytogenes estimate the survival outcomes and groups were compared with expressing OVA (day 10). Three days after the bacterial challenge, log-rank statistic. bacterial burden in spleen and liver was measured by culturing serially diluted homogenized spleen and liver on brain-heart Acknowledgments infusion PBMC agar plate. Peptide-specific IFNc-secreting cells in We thank Dr. Hao Shen for Listeria-Ova, the FACS Core Facility were measured after restimulation with SIINFEKL in at the MD Anderson for assistance with cell sorting and the the presence of Golgi-Plug for 5 h followed by intracellular NIH tetramer core facility (Atlanta, GA) for providing PBS57- staining of IFNc. In a tumor challenge model, mice vaccinated with indicated cellular vaccines were i.v. challenged with live 2 ! 105 B16-OVA 10 d after vaccination. Two weeks after the tumor loaded CD1d-tetramers. This study was funded in part by grants from the National Institutes of Health (C.D.) and an MD Anderson Interdisciplinary Research Project (C.D.). C.D. is a challenge, tumor foci in the lung were counted and OVA-specific Cancer Research Institute Investigator and a Research Trust © CD8 T 2012 cells were measured after staining Landes with PE-conjugated Fellow of Bioscience. MD Anderson Cancer Center. D.Z. is supported by OT-I tetramer in combination with FITC-conjugated anti-CD8 MD Anderson Cancer Center and a Developmental Award Ab. To test protective antitumor effect by autologous vaccine, from NCI Joe Moakley Leukemia Spore grant. The Flow aGC loaded EG-7 cells were irradiated before i.v. vaccination Cytometry Core Facility is supported by The University of (1x106 cells). At 7 d later, mice were s.c. challenged 1 ! 106 live Texas MD Anderson Cancer Center Support Grant CA16672 EG-7 cells and then the survival and tumor size of mice were (NIH). monitored. To deplete CD8 T cells, 100 mg of anti-CD8 mAb vaccinated with EG-7D/aGC oonday-7. nOn doay0, mtice wderei.v.i sNopottenrtial cionbflicts of uinteretsts ewere d.isclosed. (563.8) was i.p. injected twice (on day-3 and -1) into the mice Disclosure of Potential Conflicts Of Interest Statement References 1. Tupin E, Kinjo Y, Kronenberg M. The unique role of natural killer T cells in the response to microorganisms. Nat Rev Microbiol 2007; 5:405-17; PMID:17487145; http://dx.doi.org/10.1038/nrmicro1657 2. Fujii S, Shimizu K, Smith C, Bonifaz L, Steinman RM. Activation of natural killer T cells by alpha-galactosyl- ceramide rapidly induces the full maturation of dendritic cells in vivo and thereby acts as an adjuvant for combined CD4 and CD8 T cell immunity to a coadministered protein. 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