Modulation of p38 MAPK signaling enhances dendritic cell activation of human CD4+ Th17 responses to ovarian tumor antigen

Martin J Cannon, Hannah E Goyne, Pamela J B Stone, Laura J Macdonald, Lindsey E James, Everardo Cobos, Maurizio Chiriva-Internati, Martin J Cannon, Hannah E Goyne, Pamela J B Stone, Laura J Macdonald, Lindsey E James, Everardo Cobos, Maurizio Chiriva-Internati

Abstract

The recent finding that Th17 infiltration of ovarian tumors positively predicts patient outcomes suggests that Th17 responses play a protective role in ovarian tumor immunity. This observation has led to the question of whether Th17 cells could be induced or expanded to therapeutic advantage by tumor vaccination. In this study, we show that treatment of ovarian tumor antigen-loaded, cytokine-matured human dendritic cells (DC) with a combination of IL-15 and a p38 MAP kinase inhibitor offers potent synergy in antagonism of CD4(+) Treg induction and redirection toward CD4(+) Th17 responses that correlate with strong CD8(+) cytotoxic T lymphocyte (CTL) activation. Ovarian tumor antigen-specific CD4(+) T cells secrete high levels of IL-17 and show reduced expression of CTLA-4, PD-1, and Foxp3 following activation with IL-15/p38 inhibitor-treated DC. We further show that modulation of p38 MAPK signaling in DC is associated with reduced expression of B7-H1 (PD-L1), loss of indoleamine 2,3-dioxygenase activity, and increased phosphorylation of ERK 1/2 MAPK. These observations may allow the development of innovative DC vaccination strategies to boost Th17 immunity in ovarian cancer patients.

Conflict of interest statement

Conflict of interest Dr. Cannon is founder of DCV Technologies Inc, a biotechnology company dedicated to the clinical development of dendritic cell vaccines for the treatment of cancer. Dr. Chiriva- Internati is founder of Kiromic Inc, a biotechnology company that seeks to develop therapeutic cancer vaccines. The other authors declare no conflict of interest.

Figures

Fig. 1
Fig. 1
Functional analysis of hepsin 48–84 tumor antigen-specific CD4+ T cell lines derived by stimulation with cytokine-matured DC, IL-15-treated cytokine-matured DC, p38i-treated cytokine-matured DC or IL-15/p38i-treated, cytokine-matured DC. a CD4+ T cell expression of TNFα, IFNγ and Foxp3 following stimulation with antigen-loaded autologous LCL. CD4+ T cells cocultured with control LCL did not express TNFα or IFNγ (not shown). Representative results from two healthy subjects are shown (from a total of ten experiments). b CD8+ CTL responses against antigen-loaded autologous LCL (filled circle) or control LCL (open circle). c CD4+ T cell IL-17 expression following stimulation with antigen-loaded autologous LCL. Statistically significant differences were observed between IL-17 expression by the IL-15/p38i DC-derived CD4+ T cells and all other CD4+ T cell groups (*P ≤ 0.008). For all groups, CD4+ T cells cocultured with control LCL expressed <15 pg/ml IL-17 (not shown). LCL alone did not express IL-17
Fig. 2
Fig. 2
Costimulatory molecule expression by mature DC following treatment with IL-15 and/or p38 inhibition. a Expression of CD80, CD86, and CD83. b Expression of B7-H1 (PD-L1) and ICOS-L
Fig. 3
Fig. 3
CTLA-4 and PD-1 expression by CD4+ T cell lines derived by DC stimulation following treatment of DC with IL-15 and/or p38 inhibition
Fig. 4
Fig. 4
Phosphorylation of p38, ERK, and NF-κB following treatment of DC with IL-15 and/or p38 inhibition
Fig. 5
Fig. 5
IDOactivity inDC. Kynurenine production by cytokine-matured DC, IL-15-treated cytokine-matured DC, p38i-treated cytokine-matured DC or IL-15/p38i-treated, cytokine-matured DC, incubated with or without tryptophan (the IDO substrate). Statistically significant differences in IDO activity were observed between p38i-treated and untreated DC, and also between cytokine-matured DC and IL-15-treated cytokine-matured DC (*P ≤ 0.001 for all comparisons)
Fig. 6
Fig. 6
Summary of phenotypic and functional changes in mature DC and CD4+ T cells following DC treatment with IL-15 and/or p38 inhibition

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