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. 2021 Jan 6;9(1):e001402. doi: 10.1136/jitc-2020-001402

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© Author(s) (or their employer(s)) 2021. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.

This is an open access article distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited, appropriate credit is given, any changes made indicated, and the use is non-commercial. See http://creativecommons.org/licenses/by-nc/4.0/.

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Identification and characterization of selective HPK1 inhibitor, CompK. (A) Chemical structure of CompK. (B) Enhanced IFN-γ production in human CD8+ T cells by CompK. Cells were stimulated by anti-CD3 and anti-CD28 for 48 hours in the presence of different concentrations of CompK followed by measurement of IFN-γ release. (C) Reduction of pSLP76 in CD4+ and CD8+ T cells by CompK in human whole blood. Heparinized fresh human whole blood was treated with CompK for 60 min followed by stimulation with anti-CD3 and anti-CD28 for 15 min. Cells were stained and pSLP76 was measured by flow cytometry. (D) Increased production of IFN-γ by CompK treatment using human whole blood. Human whole blood was stimulated with anti-CD3 and anti-CD28 in the presence of different concentrations of CompK for 3 hours. Messenger RNA and complementary DNA were generated followed by quantitative PCR analysis. Studies were conducted with multiple donors; representative graphs are shown here. CompK, compound K; DMSO, dimethyl sulfoxide; IFN-γ, interferon gamma.