Department of Dermatology, Yale University School of Medicine, New Haven, CT, USA.
Department of Chemical & Environmental Engineering, Yale University School of Engineering and Applied Science, New Haven, CT, USA.
Methods Mol Biol. 2021;2255:171-186. doi: 10.1007/978-1-0716-1162-3_15.
Immunogenic cell death (ICD) is a form of regulated cell death that is capable of eliciting an immune response. In cancer, tumor cells undergoing ICD are known to emit damage associated molecular patterns (DAMPs) that are capable of recruiting and activating antigen presenting cells (APCs), which ultimately lead to the activation of an antitumor immune response. Surface translocation of intracellular chaperones such as calreticulin, release of TLR agonists such as high mobility box 1, and the secretion of type I IFN are some of the hallmark features seen in tumors succumbing to ICD. While detection of these molecules is suggestive of ICD induction, which alone does not certify that the treatment is an ICD inducer, an in vivo vaccination assay using injured tumor cells remains to be the gold standard method to functionally verify ICD. This chapter will discuss the necessary steps required to conduct an in vivo vaccination assay, focusing on the preparation of vaccine using treated tumor cells, and how these cells are then utilized in the animal model.
免疫原性细胞死亡(ICD)是一种受调控的细胞死亡形式,能够引发免疫反应。在癌症中,经历 ICD 的肿瘤细胞被认为会释放损伤相关分子模式(DAMPs),这些 DAMPs 能够招募和激活抗原呈递细胞(APCs),最终导致抗肿瘤免疫反应的激活。细胞内伴侣蛋白如钙网蛋白的表面易位、TLR 激动剂如高迁移率族蛋白 1 的释放以及 I 型 IFN 的分泌是肿瘤对 ICD 敏感时出现的一些特征性特征。虽然这些分子的检测提示 ICD 的诱导,但仅凭这一点并不能证明该治疗是 ICD 诱导剂,使用受损肿瘤细胞进行体内疫苗接种试验仍然是功能验证 ICD 的金标准方法。本章将讨论进行体内疫苗接种试验所需的必要步骤,重点介绍使用处理后的肿瘤细胞制备疫苗,以及如何在动物模型中利用这些细胞。