Immunologic monitoring of cancer vaccine trials using the ELISPOT assay

Cancer vaccines are designed to activate an immune response to tumor-specific or tumor-associated antigens expressed by the tumor. Cancer vaccines take many forms, including synthetic peptides, tumor cells and lysates, cell lines, and autologous antigen presenting cells like dendritic cells. The target antigens may be known, or "defined" in the vaccine, or unknown. In melanoma, more so than in other cancers, a large number of immunogenic "shared" antigens (tumor-specific or tumor-associated) have been identified. This allows for vaccination of groups of patients with the same vaccine, and also allows for testing for melanoma tumor immunity even when the vaccine does not include defined antigens. For the cancer vaccine field, the goal of a prognostic or predictive biomarker has yet to be achieved. However, the primary immunologic goal of any cancer vaccine is the induction (or amplification) of an immune response against the tumor, therefore the primary goal of immunologic monitoring in this setting, is testing for that response. In this chapter, we present standardized methodology from a central immunologic monitoring laboratory for melanoma cancer vaccine immune response assessment by the Enzyme-Linked Immunosorbant Spot (ELISPOT) assay. This assay allows for enumeration of antigen-specific cells in a plate format. We present the Interferon (IFN)-γ-producing lymphocyte assay, but the platform is easily adjusted to several cell types and several secreted molecules.