Research Article: Down-Regulation of the Interferon Signaling Pathway in T Lymphocytes from Patients with Metastatic Melanoma

Date Published: May 8, 2007

Publisher: Public Library of Science

Author(s): Rebecca J Critchley-Thorne, Ning Yan, Serban Nacu, Jeffrey Weber, Susan P Holmes, Peter P Lee, Franco M Marincola

Abstract: BackgroundDysfunction of the immune system has been documented in many types of cancers. The precise nature and molecular basis of immune dysfunction in the cancer state are not well defined.Methods and FindingsTo gain insights into the molecular mechanisms of immune dysfunction in cancer, gene expression profiles of pure sorted peripheral blood lymphocytes from 12 patients with melanoma were compared to 12 healthy controls. Of 25 significantly altered genes in T cells and B cells from melanoma patients, 17 are interferon (IFN)-stimulated genes. These microarray findings were further confirmed by quantitative PCR and functional responses to IFNs. The median percentage of lymphocytes that phosphorylate STAT1 in response to interferon-α was significantly reduced (Δ = 16.8%; 95% confidence interval, 0.98% to 33.35%) in melanoma patients (n = 9) compared to healthy controls (n = 9) in Phosflow analysis. The Phosflow results also identified two subgroups of patients with melanoma: IFN-responsive (33%) and low-IFN-response (66%). The defect in IFN signaling in the melanoma patient group as a whole was partially overcome at the level of expression of IFN-stimulated genes by prolonged stimulation with the high concentration of IFN-α that is achievable only in IFN therapy used in melanoma. The lowest responders to IFN-α in the Phosflow assay also showed the lowest gene expression in response to IFN-α. Finally, T cells from low-IFN-response patients exhibited functional abnormalities, including decreased expression of activation markers CD69, CD25, and CD71; TH1 cytokines interleukin-2, IFN-γ, and tumor necrosis factor α, and reduced survival following stimulation with anti-CD3/CD28 antibodies compared to controls.ConclusionsDefects in interferon signaling represent novel, dominant mechanisms of immune dysfunction in cancer. These findings may be used to design therapies to counteract immune dysfunction in melanoma and to improve cancer immunotherapy.

Partial Text: Cancer inhibits the immune system by various cellular and molecular mechanisms. Dysfunction of the immune system arises during the early stages of cancer and throughout progression to metastatic disease [1]. CD8 T lymphocytes specific for tumor-associated antigens (TAAs) are often present in the blood of cancer patients and accumulate in tumor-draining lymph nodes and in primary and metastatic tumor sites [2]. While TAA-specific CD8 T cells are elicited in the majority of patients receiving current peptide vaccines and other immunotherapies, they do not effectively control or eradicate tumors, and the presence or magnitude of these responses does not reliably correlate with clinical outcome [3]. Such cells may be specifically driven into apoptosis [4,5] or rendered nonresponsive (anergic) in vivo, preventing cytolytic responses against tumor cells and appropriate activation to stimuli. Indeed, dysfunction of TAA-specific CD8 T cells has been shown in melanoma and other cancers [2,6]. TAA-specific CD4 T cells have also been identified and are thought to help the function, persistence, and magnitude of antigen-specific CD8 T cell responses [7,8]. Current immunotherapeutic strategies are subject to the immunosuppressive effects of cancer and regulatory T cells, which likely contribute to their lack of success thus far [9–13]. The precise nature and molecular basis of immune dysfunction in the cancer state are not well defined. Elucidation of the mechanisms of immune dysfunction in cancer will allow rational design of strategies to reverse existing immune dysfunction and normalization of lymphocyte populations to improve the endogenous immune responses to cancer and to improve the efficacy of cancer immunotherapy.

Dysfunction or nonresponsiveness of the immune system may be an early event in tumor progression, while global immune suppression develops in most patients with metastatic disease [1,2]. The molecular mechanisms underlying immune dysfunction in cancer remain unclear. In this study, we studied lymphocytes from patients with metastatic melanoma at the level of gene expression using DNA microarrays to identify immune signatures that are associated with the cancer state. To resolve gene expression changes in specific lymphocyte subsets, we analyzed pure cell populations, stringently sorted by flow cytometry to allow precise analysis of each cell type. Our study included all the peripheral blood lymphocyte populations that are potentially involved in antitumor responses and may be negatively impacted by tumors, specifically CD8 T cells, CD4 T cells, B cells, and CD56dim NK cells. Inhibition of these key immune cell subsets in cancer may aid tumor progression and confound immunotherapeutic approaches.



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