A multispectral immunohistochemistry panel to investigate γδ T cells and butyrophilin molecules in the tumour microenvironment

Jessica da Gama Duarte, Luke T. Quigley, Elnaz Tavancheh, Simone Ostrouska, A. Behren
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引用次数: 0

Abstract

Conventional immunohistochemistry methods though once fundamental for the individual staining of cell markers, have now been superseded by multispectral immunohistochemistry (mIHC). mIHC enables simultaneous detection of multiple cell markers in situ using single formalin-fixed paraffin-embedded (FFPE) tissue sections. In addition to conserving patient tissue specimens, the ability to visualise more than one marker on individual cells allows for further refining of cell phenotypes, and provides insight into cell-to-cell interactions and spatial arrangements across single tissue sections. Here, a comprehensive protocol is described for the in situ interrogation of γδ T cells and phosphoantigen-presenting butyrophilin (BTN) molecules (BTN2A1 and BTN3A1) in human FFPE tissue using Opal™ tyramide signal amplification (TSA)-based mIHC. It is demonstrated that an effectively optimised Opal™-TSA 7-marker [CD3, Pan-γδ T cell receptor (TCR), granzyme B, BTN2A1, BTN3A1, tumour marker, 4’,6-diamidino-2-phenylindole (DAPI)] mIHC panel can be used to define the presence, localisation, and activation status of γδ T cells and the BTN2A1 and BTN3A1 ligands.
肿瘤微环境中γδ T细胞和嗜丁酸蛋白分子的多光谱免疫组化研究
传统的免疫组织化学方法虽然曾经是细胞标记物单独染色的基础,但现在已经被多光谱免疫组织化学(mIHC)所取代。使用单个福尔马林固定石蜡包埋(FFPE)组织切片,mIHC能够同时检测多个细胞标记物。除了保存患者组织标本外,在单个细胞上可视化多个标记的能力允许进一步完善细胞表型,并提供洞察细胞间相互作用和跨单个组织切片的空间安排。本文描述了一种基于Opal™酪酰胺信号放大(TSA)的mIHC对人FFPE组织中γδ T细胞和磷酸抗原呈递嗜丁酸蛋白(BTN)分子(BTN2A1和BTN3A1)的原位审讯的综合方案。结果表明,有效优化的Opal™-TSA 7标记物[CD3, Pan-γδ T细胞受体(TCR),颗粒酶B, BTN2A1, BTN3A1,肿瘤标记物,4 ',6-二氨基-2-苯基吲哚(DAPI)] mIHC面板可用于定义γδ T细胞和BTN2A1和BTN3A1配体的存在,定位和激活状态。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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