Ag-Specific Type 1 CD8 Effector Cells Enhance Methotrexate-Mediated Antitumor Responses by Modulating Endogenous CD49b-Expressing CD4 and CD8 T Effector Cell Subpopulations Producing IL-10

Abstract

The chemotherapeutic agent methotrexate is widely used in the treatment of breast cancer. Although its mechanism-of-action has been defined, less is known about its interaction with Ag-specific T cell-mediated antitumor responses. Type 1 CD8 T cell-mediated immune responses (Tc1) are cytolytic, produce IFN-gamma and are associated with effective antitumor responses. Using a murine transgenic TCR tumor model, we show that single-dose-treatment with methotrexate enhanced CD8-mediated type 1 antitumor responses when administered three days prior to Tc1 effector cell transfer. Co-treatment with methotrexate not only enhanced donor Tc1 cell accumulation and persistence at sites of primary tumor growth, but also promoted elevated levels of activated CD25⁺ expressing donor TIL cells. This correlated with a marked decrease in the appearance of endogenous differentiated (CD44^High) CD3/CD8/CD49b and CD3/CD4/CD49b tumor-infiltrating effector T cells at both early (Days 1–8) and late (Days 12–20) stages following treatment when compared to that of corresponding groups receiving either MTX or Tc1 cell transfer alone. Moreover, such cellular response kinetics appeared to further correlate with the down-regulation of endogenous CD4/CD44^High/CD49b effector T cells producing IL-10 and delays in tumor growth in vivo. This suggested that Ag-specific Tc1 cell transfer, in combination with chemotherapy, can enhance antitumor responses by modulating select CD49b-expressing T effector/memory cell subpopulations involved in homeostasis and immune tolerance within the tumor environment. These studies offer insight into mechanisms that enhance T cell-based immunotherapy in cancer. Supplementary materials are available for this article. Go to the publisher's online edition of Immunological Investigations for the following free supplemental resource(s): Addendum 1.

Keywords:

• Tumor immunity
• Cytokines
• Chemotherapy
• T effector/memory cells
• Tc1/Th1 cells
• IL-10
• Immunoregulatory cells

ABBREVIATIONS
TIL:	=	Tumor Infiltrating T Cells
TSA-HA:	=	HA-expressing TS/A breast adenocarcinoma
Tc1/Th1:	=	CD8+ or CD4+ T cells producing type 1 cytokines
Treg:	=	Regulatory T cells
MTX:	=	Methotrexate