Li Kelin, Vorkas Charles K, Chaudhry Ashutosh, Bell Donielle L, Willis Richard A, Rudensky Alexander, Altman John D, Glickman Michael S, Aubé Jeffrey
Division of Chemical Biology and Medicinal Chemistry, UNC Eshelman School of Pharmacy, University of North Carolina, Chapel Hill, North Carolina, United States of America.
Division of Infectious Diseases, Weill Cornell Medicine, New York, New York, United States of America.
PLoS One. 2018 Feb 5;13(2):e0191837. doi: 10.1371/journal.pone.0191837. eCollection 2018.
Mucosal-associated invariant T (MAIT) cells are an abundant class of innate T cells restricted by the MHC I-related molecule MR1. MAIT cells can recognize bacterially-derived metabolic intermediates from the riboflavin pathway presented by MR1 and are postulated to play a role in innate antibacterial immunity through production of cytokines and direct bacterial killing. MR1 tetramers, typically stabilized by the adduct of 5-amino-6-D-ribitylaminouracil (5-A-RU) and methylglyoxal (MeG), are important tools for the study of MAIT cells. A long-standing problem with 5-A-RU is that it is unstable upon storage. Herein we report an efficient synthetic approach to the HCl salt of this ligand, which has improved stability during storage. We also show that synthetic 5-A-RU•HCl produced by this method may be used in protocols for the stimulation of human MAIT cells and production of both human and mouse MR1 tetramers for MAIT cell identification.
黏膜相关恒定T（MAIT）细胞是一类丰富的天然T细胞，受主要组织相容性复合体（MHC）I类相关分子MR1的限制。MAIT细胞能够识别由MR1呈递的来自核黄素途径的细菌衍生代谢中间体，并被推测通过产生细胞因子和直接杀灭细菌在天然抗菌免疫中发挥作用。MR1四聚体通常由5-氨基-6-D-核糖基氨基尿嘧啶（5-A-RU）和甲基乙二醛（MeG）加合物稳定，是研究MAIT细胞的重要工具。5-A-RU长期存在的一个问题是其在储存时不稳定。在此，我们报告了一种合成该配体盐酸盐的有效方法，其在储存期间具有更高的稳定性。我们还表明，通过该方法产生的合成5-A-RU•HCl可用于刺激人MAIT细胞以及制备用于鉴定MAIT细胞的人和小鼠MR1四聚体的实验方案中。
