PD1-CD28 Fusion Protein Enables CD4+ T Cell Help for Adoptive T Cell Therapy in Models of Pancreatic Cancer and Non-hodgkin Lymphoma.

Felicitas Rataj,Michael Chaloupka,Stefan Endres,Simon Grassmann,Bruno L Cadilha,Fabian B T Kraus,Constanze Heise,Sebastian Kobold,Peter Duewell

doi:10.3389/fimmu.2018.01955

Abstract

Background: Interaction of the programmed death receptor 1 (PD-1) and its ligand, PD-L1, suppresses T cell activity and permits tumors to evade T cell-mediated immune surveillance. We have recently demonstrated that antigen-specific CD8+ T cells transduced with a PD1-CD28 fusion protein are protected from PD-1-mediated inhibition. We have now investigated the potential of PD1-CD28 fusion protein-transduced CD4+ T cells alone or in combination with CD8+ T cells for immunotherapy of pancreatic cancer and non-Hodgkin lymphoma.Methods: OVA-specific CD4+ and CD8+ were retrovirally transduced with the PD1-CD28 fusion protein. Cytokine release, proliferation, cytotoxic activity, and phenotype of transduced T cells were assessed in the context of Panc02-OVA (murine pancreatic cancer model) and E.G7-PD-L1 (murine T cell lymphoma model) cells.Results: Stimulation of PD1-CD28 fusion protein-transduced CD4+ T cells with anti-CD3 and recombinant PD-L1 induced specific T cell activation, as measured by IFN-y release and T cell proliferation. Coculture with Panc02-OVA or E.G7-PD-L1 tumor cells also led to specific activation of CD4+ T cells. Cytokine release and T cell proliferation was most effective when tumor cells simultaneously encountered genetically engineered CD4+ and CD8+ T cells. Synergy between both cell populations was also observed for specific tumor cell lysis. T cell cytotoxicity was mediated via granzyme B release and mediated enhanced tumor control in vivo. Transduced CD4+ and CD8+ T cells in co-culture with tumor cells developed a predominant central memory phenotype over time. Different ratios of CD4+ and CD8+ transduced T cells led to a significant increase of IFN-y and IL-2 secretion positively correlating with CD4+ T cell numbers used. Mechanistically, IL-2 and MHC-I were central to the synergistic activity of CD4+ and CD8+ T cells, since neutralization of IL-2 prevented the crosstalk between these cell populations.Conclusion: PD1-CD28 fusion protein-transduced CD4+ T cells significantly improved anti-tumoral effect of fusion protein-transduced CD8+ T cells. Thus, our results indicate that PD1-CD28 fusion protein-transduced CD4+ T cells have the potential to overcome the PD-1-PD-L1 immunosuppressive axis in pancreatic cancer and non-Hodgkin lymphoma.

Highlights

Cytotoxic T cells recognize tumor antigens presented on major histocompatibility complex-1 (MHC-I)
Our results indicate the potential of PD1-CD28 fusion protein-transduced CD4+ T cells to further improve adoptive T cell therapy (ACT)
PTMtransduced and untransduced T cells were stimulated with anti-CD3 antibody, anti-CD3 antibody and recombinant PDL1 or anti-CD3 antibody and anti-CD28 antibody for 48 h

Summary

Introduction

Cytotoxic T cells recognize tumor antigens presented on major histocompatibility complex-1 (MHC-I). After binding to the tumor antigen in the context of MHC, T cells are activated, which results in the secretion of cytotoxic factors and target cell lysis [1, 2]. This concept is utilized therapeutically for adoptive T cell therapy (ACT). When being transduced for tumor specificity both cell types are being modified and in the case of CAR T cells, both cell populations are thought to be therapeutically relevant [9]. We have recently demonstrated that antigen-specific CD8+ T cells transduced with a PD1-CD28 fusion protein are protected from PD-1-mediated inhibition. We have investigated the potential of PD1-CD28 fusion protein-transduced CD4+ T cells alone or in combination with CD8+ T cells for immunotherapy of pancreatic cancer and non-Hodgkin lymphoma

Methods

Results

Conclusion