Enhancement of Intracellular Signaling in T Cell Receptor-transduced T Cells

2020 Fiscal Year Final Research Report

• Project/Area Number: 18K08351
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 54010:Hematology and medical oncology-related
• Research Institution: Nagoya University
• Principal Investigator: Terakura Seitaro 名古屋大学, 医学部附属病院, 講師 (40625141)
• Project Period (FY): 2018-04-01 – 2021-03-31
• Keywords: 遺伝子導入T細胞療法 / アダプター分子 / Tリンパ球

Outline of Final Research Achievements

In this study, we attempted to develop a novel vector that enables gene transfer of TCR and adapter molecule (ATAM) into T cells by a single gene transfer. One virus vector method (1vv method) using all-in-one vector and two virus vector (2vv) method using two vectors used in the previous study showed that the expression of ATAM was about 3 times higher in 2vv method. The expression of ATAM was about 3-fold higher in the 2vv method. Comparison of the function of the TCR-T cells generated by the 2vv method showed that the cells generated by the 2vv method proliferated significantly better after antigen stimulation, and the intracellular signals after stimulation showed that the signals of phosphorylated Erk and p38 were significantly enhanced in the TCR-T cells generated by the 2vv method. The results showed that it is necessary to transfer three times more ATAM than TCR.

Free Research Field

血液学

Academic Significance and Societal Importance of the Research Achievements

T細胞レセプター遺伝子と新規に開発したアダプター分子を１回の遺伝子導入によりT細胞内に遺伝子導入可能とする新規ベクターの開発を試みた。ひとつのベクターで遺伝し導入する1 virus vector法（1vv法)と二つのウイルスベクターを用いる方法 (2 virus vector法, 2vv法)を比較したところ、ウェスタンブロット法でも、フローサイト メトリー法でも2vv法でアダプター分子の発現が約3倍程度高く、2vv法で作成したTCR-Tの方が有意に抗原刺激後の増殖が有利であった。TCRよりも3倍以上多くのアダプター分子を遺伝子導入する必要があることが示された。