评估癌症疫苗在临床前癌症模型中的体内免疫原性和治疗疗效的适当方法对于克服当前癌症疫苗的限制并增强这种有前途的免疫治疗策略的临床适用性至关重要。 特别是需要可用于表征内源性肿瘤抗原的T细胞反应的方法来评估疫苗效力并改善抗原制剂。 此外，多参数分析用于深入地表征肿瘤诱导和治疗诱导的免疫调节对于设计基于机制的联合免疫治疗非常重要。 本文描述了一种多功能流式细胞术，用于评估基于相关小鼠品系的相关肿瘤表位的短期体外重刺激后肿瘤抗原特异性CD4 +和CD8 + T细胞反应的多种细胞因子产生能力。我们还报告了开发和应用两个21色流式细胞术检测面板，全面表征小鼠的T细胞和自然杀伤细胞衰竭和记忆表型，特别关注临床前癌症模型。版权所有©2023 Moi、Zeng、Minnie、Bhatt、Wood、Sester、Mazzieri和Dolcetti。

Suitable methods to assess in vivo immunogenicity and therapeutic efficacy of cancer vaccines in preclinical cancer models are critical to overcome current limitations of cancer vaccines and enhance the clinical applicability of this promising immunotherapeutic strategy. In particular, availability of methods allowing the characterization of T cell responses to endogenous tumor antigens is required to assess vaccine potency and improve the antigen formulation. Moreover, multiparametric assays to deeply characterize tumor-induced and therapy-induced immune modulation are relevant to design mechanism-based combination immunotherapies. Here we describe a versatile multiparametric flow cytometry method to assess the polyfunctionality of tumor antigen-specific CD4+ and CD8+ T cell responses based on their production of multiple cytokines after short-term ex vivo restimulation with relevant tumor epitopes of the most common mouse strains. We also report the development and application of two 21-color flow cytometry panels allowing a comprehensive characterization of T cell and natural killer cell exhaustion and memory phenotypes in mice with a particular focus on preclinical cancer models.Copyright © 2023 Moi, Zeng, Minnie, Bhatt, Wood, Sester, Mazzieri and Dolcetti.