This protocol describes how to induce large numbers of tumor-specific cytotoxic T cells (CTLs) in the spleens and lymph nodes of mice receiving dendritic cell (DC) vaccines and how to modulate tumor microenvironments (TMEs) to ensure effective homing of the vaccination-induced CTLs to tumor tissues. We also describe how to evaluate the numbers of tumor-specific CTLs within tumors. The protocol contains detailed information describing how to generate a specialized DC vaccine with augmented ability to induce tumor-specific CTLs. We also describe methods to modulate the production of chemokines in the TME and show how to quantify tumor-specific CTLs in the lymphoid organs and tumor tissues of mice receiving different treatments. The combined experimental procedure, including tumor implantation, DC vaccine generation, chemokine-modulating (CKM) approaches, and the analyses of tumor-specific systemic and intratumoral immunity is performed over 30-40 d. The presented ELISpot-based ex vivo CTL assay takes 6 h to set up and 5 h to develop. In contrast to other methods of evaluating tumor-specific immunity in tumor tissues, our approach allows detection of intratumoral T-cell responses to nonmanipulated weakly immunogenic cancers. This detection method can be performed using basic laboratory skills, and facilitates the development and preclinical evaluation of new immunotherapies.

中文翻译：

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通过树突状细胞疫苗和趋化因子调节剂促进肿瘤特异性T细胞在肿瘤组织中的积累。


该方案描述了如何在接受树突状细胞（DC）疫苗的小鼠的脾脏和淋巴结中诱导大量肿瘤特异性细胞毒性T细胞（CTL），以及如何调节肿瘤微环境（TME）以确保疫苗有效归巢 -诱导CTLs到达肿瘤组织。我们还描述了如何评估肿瘤内肿瘤特异性 CTL 的数量。该协议包含详细信息，描述如何生成具有增强诱导肿瘤特异性 CTL 能力的专用 DC 疫苗。我们还描述了调节 TME 中趋化因子产生的方法，并展示了如何量化接受不同治疗的小鼠的淋巴器官和肿瘤组织中的肿瘤特异性 CTL。组合实验程序，包括肿瘤植入、DC 疫苗生成、趋化因子调节 (CKM) 方法以及肿瘤特异性全身和瘤内免疫分析，持续 30-40 天。所提出的基于 ELISpot 的离体 CTL 测定需要 6 小时来建立，5 小时来开发。与评估肿瘤组织中肿瘤特异性免疫的其他方法相比，我们的方法可以检测肿瘤内 T 细胞对非操纵的弱免疫原性癌症的反应。这种检测方法可以使用基本的实验室技能进行，并有助于新免疫疗法的开发和临床前评估。