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Localized expression of GITR-L in the tumor microenvironment promotes CD8+ T cell dependent anti-tumor immunity

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Abstract

The systemic administration of an agonist antibody against glucocorticoid-induced tumor necrosis factor receptor related (GITR) protein has been shown to be effective in overcoming immune tolerance and promoting tumor rejection in a variety of murine tumor models. However, little is known regarding the functional consequence of ligation of GITR with its natural ligand (GITR-L) in the context of regulatory T cell (Treg) suppression in vivo. To determine the mechanism of GITR-L action in vivo, we generated a panel of tumor cell clones that express varying levels of GITR-L. The ectopic expression of GITR-L on the tumor cell surface was sufficient to enhance anti-tumor immunity and delay tumor growth in syngeneic BALB/c mice. Within the range examined, the extent of anti-tumor activity in vivo did not correlate with the level of GITR-L expression, as all clones tested exhibited a similar delay in tumor growth. The localized expression of GITR-L on tumor cells led to a significant increase in CD8+ T cell infiltration compared to the levels seen in control tumors. The increased proportion of CD8+ T cells was only observed locally at the tumor site and was not seen in the tumor draining lymph node. Depletion studies showed that CD8+ T cells, but not CD4+ T cells, were required for GITR-L mediated protection against tumor growth. These studies demonstrate that signaling between GITR-L and GITR in the tumor microenvironment promotes the infiltration of CD8+ T cells, which are essential for controlling tumor growth.

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Abbreviations

GITR:

Glucocorticoid-induced TNF receptor related

GITR-L:

GITR ligand

Tregs:

Regulatory T cells

ECD:

Extracellular domain

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Acknowledgments

We thank Dr. Koteswara R. Chintalacharuvu, Caiyun Xuan, and Sepideh Afshar for critically reading the manuscript. We are also grateful to Letitia Wims for help with protein purification and Dr. B. S. Kwon for generously providing the cDNA of GITR-L. Flow cytometry was performed in the UCLA Jonsson Comprehensive Cancer Center (JCCC) and Center for AIDS Research Flow Cytometry Core Facility that is supported by National Institutes of Health awards CA-16042 and AI-28697, and by the JCCC, the UCLA AIDS Institute, the David Geffen School of Medicine at UCLA, and the UCLA Chancellor’s Office. J. S. Cho was supported in part by NIH Tumor Immunology training grant 5 T32 CA009120: 32.

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  1. Department of Microbiology, Immunology and Molecular Genetics, University of California Los Angeles, BSRB 247, 615 Charles E. Young Dr. South, Los Angeles, CA, 90095-1570, USA

    John S. Cho, Jeffrey V. Hsu & Sherie L. Morrison

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  1. John S. Cho
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Correspondence to Sherie L. Morrison.

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Cho, J.S., Hsu, J.V. & Morrison, S.L. Localized expression of GITR-L in the tumor microenvironment promotes CD8+ T cell dependent anti-tumor immunity. Cancer Immunol Immunother 58, 1057–1069 (2009). https://doi.org/10.1007/s00262-008-0622-2

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