Methods for Analyzing Immune Infiltrates and Spatial Organization Within the Melanoma Microenvironment Using...

Immunotherapies improve survival in patients with melanoma by enhancing the immune system's ability to recognize and eliminate tumor cells. Understanding the immunological effects of these therapies requires evaluation of both changes in lymphocyte composition and activation within the tumor microenvironment (TME) as well as the spatial organization of these cells. Increased lymphocyte infiltration and distinct patterns of immune cell distribution within the TME have been associated with improved survival and enhanced response to immunotherapy in melanoma. However, relatively few methodologies currently enable detailed characterization of both the structural organization and immunological features of tumor-infiltrating lymphocytes. In this chapter, we describe multiplex immunofluorescence staining approaches for the identification, characterization, and quantification of tumor-infiltrating lymphocytes in melanoma.