原位杂交
生物
免疫分型
T细胞受体
免疫组织化学
分子生物学
T细胞
流式细胞术
荧光原位杂交
病理
免疫学
医学
免疫系统
信使核糖核酸
基因
生物化学
染色体
作者
Jocelyn H. Wright,Liupei Huang,Stephanie Weaver,L. Diego Archila,Megan S. McAfee,Alexandre V. Hirayama,Aude G. Chapuis,Marie Bleakley,Anthony Rongvaux,Cameron J. Turtle,R. Savanh Chanthaphavong,Jean S. Campbell,Robert H. Pierce
标识
DOI:10.1016/j.jim.2020.112955
摘要
Identifying engineered T cells in situ is important to understand the location, persistence, and phenotype of these cells in patients after adoptive T cell therapy. While engineered cells are routinely characterized in fresh tissue or blood from patients by flow cytometry, it is difficult to distinguish them from endogenous cells in formalin-fixed, paraffin-embedded (FFPE) tissue biopsies. To overcome this limitation, we have developed a method for characterizing engineered T cells in fixed tissue using in situ hybridization (ISH) to the woodchuck hepatitis post-transcriptional regulatory element (WPRE) common in many lentiviral vectors used to transduce chimeric antigen receptor T (CAR-T) and T cell receptor T (TCR-T) cells, coupled with alternative permeabilization conditions that allows subsequent multiplex immunohistochemical (mIHC) staining within the same image. This new method provides the ability to mark the cells by ISH, and simultaneously stain for cell-associated proteins to immunophenotype CAR/TCR modified T cells within tumors, as well as assess potential roles of these cells in on-target/off-tumor toxicity in other tissue.
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