Selection of highly responsive T cell receptors by an analysis combining the expression of multiple markers |
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Authors: | My Thi Viet Ha Hiroshi Hamana Kiyomi Shitaoka Abdul Hayee Eiji Kobayashi Toshiaki Yoshikawa Tetsuya Nakatsura Reiko Saikawa Eri Sato Mitsujiro Osawa Yasumichi Hitoshi Tung Son Dang Tatsuhiko Ozawa Hiroyuki Kishi |
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Affiliation: | 1. Department of Immunology, Faculty of Medicine, Academic Assembly, University of Toyama, Toyama, Japan;2. Division of Cancer Immunotherapy, Exploratory Oncology Research & Clinical Trial Center, National Cancer Center, Kashiwa, Japan;3. Thyas Co., Ltd., Kyoto, Japan;4. Department of Pathology, Faculty of Medicine, Academic Assembly, University of Toyama, Toyama, Japan |
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Abstract: | The clinical success of T cell receptor (TCR) gene–transduced T (TCR-T) cell therapy is expected as one of the next-generation immunotherapies for cancer, in which the selection of TCRs with high functional avidity (high-functional TCRs) is important. One widely used approach to select high-functional TCRs is a comparison of the EC50 values of TCRs, which involves laborious experiments. Therefore, the establishment of a simpler method to select high-functional TCRs is desired. We herein attempted to establish a simple method to select high-functional TCRs based on the expression of T cell activation markers using the mouse T cell line BW5147.3 (BW). We examined relationships between the EC50 values of TCRs in interleukin-2 production and the expression levels of TCR activation markers on BW cells. In TCR-expressing BW cells stimulated with antigenic peptides, the CD69, CD137, and PD-1 expression was differentially induced by various doses of peptides. An analysis of TCRs derived from the tumor-infiltrating lymphocytes of murine melanoma and peripheral blood T cells of hepatocellular carcinoma patients treated with a peptide vaccination revealed that an analysis combining CD69, CD137, and PD-1 expression levels in BW cells stimulated with a single dose of an antigenic peptide selected high-functional TCRs with functional avidity assessed by EC50 values. Our method facilitates the section of high-functional TCRs among tumor-reacting TCRs, which will promote TCR-T cell therapy. The stimulation of BW cells expressing objective TCRs with a single dose of antigenic peptides and analysis combining the expression of CD69, CD137, and PD-1 allows us to select highly responsive TCRs. |
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Keywords: | activation marker functional avidity T cell line T cell receptor TCR-T cells |
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