Immunospot Assessment of T-Cell Responses in Preclinical Tumor Models with Undefined Target Antigens

Abstract

Assessment of functional tumor-specific T-cell responses in preclinical tumor models represents an important tool for successful translation of new immunotherapies to clinics. Usually, it requires a known tumor antigen target. Here, we describe the method to detect tumor-specific T cell after immunotherapies without a known antigen. Splenocytes, lymph node immune cells, or PBMCs are isolated from treated mice and stimulated with relevant tumor cells ex vivo before immunospot analysis of Granzyme B and interferon γ-positive T cells. The method is especially valuable for monitoring tumor-specific T cells after vaccination with various whole tumor vaccines or after in situ vaccination and other antigen agnostic immunotherapies, where no specific antigens are used.

Keywords: Antigen agnostic immunotherapy; FluoroSpot; Granzyme B; Immunospot; Interferon γ; T cells; Tumor-specific; Undefined target antigens.