Enhancing DC cancer vaccine by allogeneic MHC class II expression and Treg depletion
Noriko Seishima, William Becker, Purevdorj B. Olkhanud, Hoyoung M. Maeng, Miguel A. Lopez-Lago, William V. Williams, Jay A. Berzofsky
Noriko Seishima, William Becker, Purevdorj B. Olkhanud, Hoyoung M. Maeng, Miguel A. Lopez-Lago, William V. Williams, Jay A. Berzofsky
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Research Article Immunology Oncology

Enhancing DC cancer vaccine by allogeneic MHC class II expression and Treg depletion

Abstract

We assessed the therapeutic efficacy of a semiallogeneic dendritic cell (DC) vaccine in comparison to a syngeneic one for suppression of B16-F10 and TC-1 tumors. Syngeneic bone marrow–derived DCs (BMDCs) were generated from C57BL/6J mice and semiallogeneic BMDCs with a mutation in either MHC class I or II were generated from B6.C-H2-Kbm1/ByJ or B6(C)-H2-Ab1bm12/KhEgJ mice, respectively. We demonstrated in vivo and in vitro that the MHC class II semiallogeneic BMDC vaccine had superior efficacy over the syngeneic and the MHC class I semiallogeneic BMDC vaccine, providing allogeneic CD4+ T cell help to enhance the antitumor CD8+ T cell response through allogeneic stimulation by the mutant MHC class II molecules. We discovered that this help was induced only at an early stage of tumor growth and at a later stage of tumor growth; combining our BMDC vaccine with Treg depletion enhanced tumor suppression. We demonstrated the improved efficacy of a semiallogeneic BMDC vaccine that kept tumor-peptide presentation intact on syngeneic MHC class I molecules so that mutant MHC class II could provide allogeneic help. This strategy should enable promising new DC-based cancer immunotherapies, offering an alternative to autologous DC vaccines by incorporating allogenicity as an adjuvant.

Authors

Noriko Seishima, William Becker, Purevdorj B. Olkhanud, Hoyoung M. Maeng, Miguel A. Lopez-Lago, William V. Williams, Jay A. Berzofsky

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Figure 3

The E743–77 peptide–pulsed MHC class II semiallogeneic mature BMDCs activate allogeneic CD4+ T cells through allogeneic MHC class II to support E7-dependent CD8+ T cell responses in TC-1 tumor–bearing mice.

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The E743–77 peptide–pulsed MHC class II semiallogeneic mature BMDCs acti...
(A and B) Flow cytometric analysis of intracellular staining of IFN-γ in splenic CD8+ or CD4+ T cells stimulated with either syngeneic WT or MHC class II semiallogeneic bm12 mature BMDCs (mBMDCs) with or without E7 peptide pulsing. (A) CD8+ T cells and (B) CD4+ T cells were cocultured with indicated mBMDCs in 200 μL/well of 96-well round-bottom plates for 18 hours and then 6 hours with brefeldin A. (C) IFN-γ production of splenic CD8+ T cells and/or CD4+ T cells from TC-1 tumor–bearing mice stimulated with either WT or bm12 mBMDCs with or without peptide pulsing by ELISA. CD8+ T cells alone, CD4+ T cells alone, or a mixture of CD8+ T cells and CD4+ T cells were cocultured with indicated mBMDCs in 200 μL/well in 96-well round-bottom plates for 72 hours. (D) The WT or bm12 BMDCs blocked with anti–MHC class II antibody for 1 hour were subjected to flow cytometric analysis. The histograms indicate the change in I-A/I-E expression on each BMDC and the bar graphs indicate the median fluorescence intensity (MFI) of I-A/I-E with or without MHC II blocking and fluorescence minus one (FMO) control for the anti–MHC class II antibody. (E) Contrast values of IFN-γ production in the supernatants with those of C with MHC II blocked. In A–C, and E, 2 × 105 isolated CD8+ T cells or 2 × 105 isolated CD4+ T cells were cultured alone or mixed with 1 × 105 indicated mBMDCs. Data are presented as the mean ± SEM and represent 3 independent experiments, n = 4–5 per group. Statistical analysis was performed using 1-way ANOVA test with post hoc Tukey’s multiple-comparison correction. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001.