Macchi Chiara, Moregola Annalisa, Norata Giuseppe Danilo, Ruscica Massimiliano
Department of Pharmacological and Biomolecular Sciences "Rodolfo Paoletti," Università degli Studi di Milano, 20133 Milan, Italy.
Department of Pharmacological and Biomolecular Sciences "Rodolfo Paoletti," Università degli Studi di Milano, 20133 Milan, Italy; SISA Center for the Study of Atherosclerosis, Bassini Hospital, Via M. Gorki 50, Cinisello Balsamo, 20092 Milan, Italy.
STAR Protoc. 2023 May 19;4(2):102301. doi: 10.1016/j.xpro.2023.102301.
The infiltration of activated T cells, such as CD8 effector, in metabolic tissues plays a crucial role for the initiation and propagation of obesity-induced inflammation. Given the pivotal role of lactate transporter monocarboxylate transporter 1 (MCT1) in immune cell activation, we present a protocol for the isolation and activation of CD8 T lymphocytes selectively lacking MCT1. We describe steps for the induction of adipocyte differentiation, CD8 T isolation and activation, and adipocyte-CD8 T cell co-culture. We then detail qPCR analysis on differentiated adipocytes. For complete details on the use and execution of this protocol, please refer to Macchi et al..
活化T细胞（如CD8效应细胞）在代谢组织中的浸润，对肥胖诱导炎症的起始和传播起着关键作用。鉴于乳酸转运体单羧酸转运体1（MCT1）在免疫细胞活化中的关键作用，我们提出了一种用于分离和激活选择性缺乏MCT1的CD8 T淋巴细胞的方案。我们描述了诱导脂肪细胞分化、CD8 T细胞分离和激活以及脂肪细胞与CD8 T细胞共培养的步骤。然后，我们详细介绍了对分化脂肪细胞的qPCR分析。有关本方案使用和执行的完整详细信息，请参考Macchi等人的研究。
