Proc Natl Acad Sci U S A. 2024 Dec 03. 121(49): e2414792121
Mitchell P McInerney,
Wael Awad,
Michael N T Souter,
Yang Kang,
Carl J H Wang,
Kean Chan Yew Poa,
Mohamed R Abdelaal,
Ngoc H Le,
Chloe M Shepherd,
Conor McNeice,
Lucy J Meehan,
Adam G Nelson,
Jeremy M Raynes,
Jeffrey Y W Mak,
James McCluskey,
Zhenjun Chen,
Ching-Seng Ang,
David P Fairlie,
Jérôme Le Nours,
Patricia T Illing,
Jamie Rossjohn,
Anthony W Purcell.
The major histocompatibility complex class I related protein (MR1) presents microbially derived vitamin B2 precursors to mucosal-associated invariant T (MAIT) cells. MR1 can also present other metabolites to activate MR1-restricted T cells expressing more diverse T cell receptors (TCRs), some with anti-tumor reactivity. However, knowledge of the range of the antigen(s) that can activate diverse MR1-reactive T cells remains incomplete. Here, we identify pyridoxal (vitamin B6) as a naturally presented MR1 ligand using unbiased mass spectrometry analyses of MR1-bound metabolites. Pyridoxal, and the related compound, pyridoxal 5-phosphate bound to MR1 and enabled cell surface upregulation of wild type MR1*01 and MR1 expressing the Arg9His polymorphism associated with the MR1*04 allotype in a manner dependent on Lys43-mediated Schiff-base formation. Crystal structures of MR1*01 in complex with pyridoxal and pyridoxal 5-phosphate showed how these ligands were accommodated within the A-pocket of MR1. T cell lines transduced with the 7.G5 TCR, which has reported "pan-cancer" specificity, were specifically activated by pyridoxal presented by antigen-presenting cells expressing MR1*01 and MR1 allotypes bearing the less common Arg9His polymorphism. 7.G5 T cells also recognized, to a lesser extent, pyridoxal 5-phosphate and, importantly, recognition of both vitamers was blocked by an anti-MR1 antibody. 7.G5 TCR reactivity toward pyridoxal was enhanced when presented by the Arg9His polymorphism-bearing MR1 allotypes. Vitamin B6, and vitamers thereof, have been associated with various cancers, and here we describe a link between this ligand, MR1, and its allomorphs, and the pan-cancer 7.G5 TCR. This work identifies an MR1 ligand that can activate a diverse MR1-restricted TCR.
Keywords: MHC class I-related molecule (MR1); T cell receptor; mass spectrometry; metabolite; structural biology