title: Effects of Malignant Melanoma Initiating Cells on T-Cell Activation.
abstract: Although human malignant melanoma is a highly immunogenic cancer, both the endogenous antitumor immune response and melanoma immunotherapy often fail to control neoplastic progression. Accordingly, characterizing melanoma cell subsets capable of evading antitumor immunity could unravel optimized treatment strategies that might reduce morbidity and mortality from melanoma. By virtue of their preferential capacity to modulate antitumor immune responses and drive inexorable tumor growth and progression, malignant melanoma-initiating cells (MMICs) warrant closer investigation to further elucidate the cellular and molecular mechanisms underlying melanoma immune evasion and immunotherapy resistance. Here we describe methodologies that enable the characterization of immunoregulatory effects of purified MMICs versus melanoma bulk populations in coculture with syngeneic or allogeneic lymphocytes, using [3H]thymidine incorporation, enzyme-linked immunosorbent spot (ELISPOT), or ELISA assays. These assays were traditionally developed to analyze alloimmune processes and we successfully adapted them for the study of tumor-mediated immunomodulatory functions.
authors.affiliation: Harvard Skin Disease Research Center, Department of Dermatology, Brigham and Women's Hospital, Harvard Medical School, Boston, MA, USA.|Transplantation Research Program, Division of Nephrology, Children's Hospital Boston, Harvard Medical School, 300 Longwood Avenue, Boston, MA, 02115, USA.
authors.forename: Tobias
authors.initials: T
authors.lastname: Schatton
authors.affiliation: Transplantation Research Program, Division of Nephrology, Children's Hospital Boston, Harvard Medical School, 300 Longwood Avenue, Boston, MA, 02115, USA.|Center of Integrated Oncology (CIO) Cologne-Bonn, Department of Internal Medicine III, University Hospital of Bonn, Bonn, Germany.
authors.forename: Ute
authors.initials: U
authors.lastname: Schütte
authors.affiliation: Harvard Skin Disease Research Center, Department of Dermatology, Brigham and Women's Hospital, Harvard Medical School, Boston, MA, USA. markus.frank@childrens.harvard.edu.|Transplantation Research Program, Division of Nephrology, Children's Hospital Boston, Harvard Medical School, 300 Longwood Avenue, Boston, MA, 02115, USA. markus.frank@childrens.harvard.edu.
authors.forename: Markus H
authors.initials: MH
authors.lastname: Frank
keywords: Antitumor immunity; Cancer stem cell; Cytokines; ELISA; ELISPOT; Immune escape; Immunology; Immunomodulation; Immunoregulatory; Melanoma; PBMC; Proliferation; T cell activation; T effector cells; Tumor initiating cell; [3H]thymidine incorporation