Clark Kolin M, Wang Qiankun, Shan Liang
Division of Infectious Diseases, Department of Medicine, Washington University School of Medicine, Saint Louis, MO, USA.
Andrew M. and Jane M. Bursky Center for Human Immunology and Immunotherapy Programs, Washington University School of Medicine, Saint Louis, MO, USA.
Methods Mol Biol. 2023;2641:67-79. doi: 10.1007/978-1-0716-3040-2_6.
The pattern recognition receptor CARD8 is an inflammasome sensor for intracellular HIV-1 protease activity. Previously, the only method for studying the CARD8 inflammasome has been through utilizing DPP8/DPP9 inhibitors including Val-boroPro (VbP) to modestly and nonspecifically activate the CARD8 inflammasome. The identification of HIV-1 protease as a target for sensing by CARD8 has opened the door for a new method of studying the underlying mechanism of CARD8 inflammasome activation. Additionally, triggering the CARD8 inflammasome offers a promising strategy for reducing HIV-1 latent reservoirs. Here we describe the methods to study CARD8 sensing of HIV-1 protease activity through non-nucleoside reverse transcriptase inhibitor (NNRTI)-mediated pyroptosis of HIV-1-infected immune cells and through an HIV and CARD8 co-transfection model.
模式识别受体CARD8是一种用于检测细胞内HIV-1蛋白酶活性的炎性小体传感器。此前，研究CARD8炎性小体的唯一方法是利用包括缬氨酸-硼替佐米（VbP）在内的DPP8/DPP9抑制剂来适度且非特异性地激活CARD8炎性小体。HIV-1蛋白酶被鉴定为CARD8的传感靶点，这为研究CARD8炎性小体激活的潜在机制开辟了一种新方法。此外，触发CARD8炎性小体为减少HIV-1潜伏库提供了一种有前景的策略。在此，我们描述了通过非核苷逆转录酶抑制剂（NNRTI）介导的HIV-1感染免疫细胞的焦亡以及通过HIV与CARD8共转染模型来研究CARD8对HIV-1蛋白酶活性的传感方法。
