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Author Notes:

E-mail: jaube@unc.edu

We thank Phil Wong, Erika Ritter, and Matthew Adamow of the MSKCC Immune Monitoring Core for their expert consultation during the development of the MAIT cell activation assay.

We also acknowledge UNC Department of Chemistry Mass Spectrometry Core Laboratory for HR-MS analysis.

All relevant data are within the paper and its Supporting Information files.

The authors have declared that no competing interests exist.

Subjects:

Research Funding:

Support from the National Institute of Allergy and Infectious Disease (niaid.nih.gov) is gratefully acknowledged: U19 AI11143, Carl Nathan, PI (supported work by KL, CKV, MSG, and JA); U19 AI111211, Henry M. Blumberg, PI (supported work by DLB, RAW, JDA); HHSN 272201300006C (supported work by DLB, RAW, JDA); and T32AI007613-18, Roy M. Gulick, PI (supported CKV).

In addition support from the National Cancer Institute (nci.nih.gov) is acknowledged, through P30 CA008748 (supported AC and AR).

The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Keywords:

  • Science & Technology
  • Multidisciplinary Sciences
  • Science & Technology - Other Topics
  • INVARIANT T-CELLS
  • GROUP B GALACTOSYLTRANSFERASE
  • MAIT CELLS
  • RIBOFLAVIN SYNTHASE
  • RECEPTOR HETEROGENEITY
  • IMMUNE-RESPONSE
  • BIOSYNTHESIS
  • TUBERCULOSIS
  • SUBSTRATE
  • ENZYME

Synthesis, stabilization, and characterization of the MR1 ligand precursor 5-amino-6-D-ribitylaminouracil (5-A-RU)

Journal Title:

PLoS ONE

Volume:

Volume 13, Number 2

Publisher:

, Pages e0191837-e0191837

Type of Work:

Article | Final Publisher PDF

Abstract:

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-ribitylaminour-acil (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.

Copyright information:

© 2018 Li et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

This is an Open Access work distributed under the terms of the Creative Commons Attribution 4.0 International License (https://creativecommons.org/licenses/by/4.0/).
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