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Studies have shown that MR1 plays a role in microbial infection, inflammation, and tumor immunity. The antigens it presents include metabolites of microbial and self-origin as well as small-molecule drugs and form stable complexes with MR1 that are displayed on the cell surface to activate T cells. However, unlike classical MHC I and II molecules, the fundamental biology of MR1 remains poorly understood, particularly the mechanisms governing antigen loading and intracellular trafficking. This knowledge gap is largely due to the lack of molecular tools available to precisely manipulate MR1 function. In this study, we describe a high-affinity (1.6\u2009nM KD) anti-MR1 nanobody, MR1Nb1. We characterize the binding of this nanobody including affinity by ELISA and kinetics by BLI. Crucially, we map the binding epitope of MR1Nb1 on MR1 by HDX-MS, providing key insights into the mechanism through which it blocks MR1T cell activation. In functional assays MR1Nb1 effectively and specifically blocks MR1T cell activation by cells infected with M. tuberculosis or treated with M. smegmatis supernatant or the synthetic ligand deazalumazine. MR1Nb1 further stains MR1-ligand complexes on the cell surface in a flow cytometry assay. This nanobody represents a unique and versatile tool for the field, as it can be produced inexpensively and expressed intracellularly within antigen presenting cells. Hence, our study provides a powerful new molecular probe for dissecting the mechanistic underpinnings of MR1 biology and uncover its broader roles in immunity.<\/jats:p>","DOI":"10.1093\/jimmun\/vkag094","type":"journal-article","created":{"date-parts":[[2026,7,2]],"date-time":"2026-07-02T17:05:17Z","timestamp":1783011917000},"source":"Crossref","is-referenced-by-count":0,"title":["An MR1-specific nanobody capable of blocking MR1T cell activation"],"prefix":"10.1093","volume":"215","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2533-7668","authenticated-orcid":false,"given":"Timothy A","family":"Bates","sequence":"first","affiliation":[{"name":"Department of Molecular Microbiology and Immunology, Oregon Health & Sciences University , Portland, OR,","place":["United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4217-3095","authenticated-orcid":false,"given":"Corinna 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