Saha Dipongkor, Rabkin Samuel D
Department of Immunotherapeutics and Biotechnology, School of Pharmacy, Texas Tech University Health Sciences Center, Abilene, TX, USA.
Department of Neurosurgery, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA.
Methods Mol Biol. 2020;2058:179-190. doi: 10.1007/978-1-4939-9794-7_11.
Immunohistochemistry (IHC) is an integral laboratory staining technique, which is used for the detection of immune cells in mouse/human tissues or tumors. Oncolytic herpes simplex virus (oHSV) treatment or virotherapy of solid tumors results in antitumor immune responses and infiltration of a variety of immune cells into the tumor. Here, we describe a step-by-step chromogen/substrate-based single- and dual-color IHC protocol to stain immune cells in formalin-fixed, paraffin-embedded mouse glioblastoma (GBM) brain tumor sections after oHSV virotherapy. Tumor sections are deparaffinized with xylene, then gradually rehydrated using ethanol, followed by heat-mediated antigen retrieval using appropriate buffers. Tumor sections are incubated with primary antibodies, which detect a specific immune cell antigen, then incubated with peroxidase- or phosphatase-labeled secondary antibodies, followed by incubation with a color-producing substrate and color visualization (of immune cells) by light microscopy. The protocol described herein is also applicable to detect immune cells in other mouse and human tumors or organs after other forms of immunotherapy.
免疫组织化学（IHC）是一种不可或缺的实验室染色技术，用于检测小鼠/人体组织或肿瘤中的免疫细胞。溶瘤单纯疱疹病毒（oHSV）治疗或实体瘤的病毒疗法会引发抗肿瘤免疫反应，并使多种免疫细胞浸润到肿瘤中。在此，我们描述了一种基于显色剂/底物的单双色免疫组化分步方案，用于在oHSV病毒疗法后对福尔马林固定、石蜡包埋的小鼠胶质母细胞瘤（GBM）脑肿瘤切片中的免疫细胞进行染色。肿瘤切片先用二甲苯脱蜡，然后用乙醇逐步复水，接着使用适当的缓冲液进行热介导的抗原修复。肿瘤切片与检测特定免疫细胞抗原的一抗孵育，然后与过氧化物酶或磷酸酶标记的二抗孵育，随后与显色底物孵育，并通过光学显微镜观察（免疫细胞的）颜色显现。本文所述方案也适用于在其他形式的免疫治疗后检测其他小鼠和人类肿瘤或器官中的免疫细胞。
