MAIT cell-MR1 reactivity is highly conserved across multiple divergent species

Matthew D Edmans; Timothy K Connelley; Sophie Morgan; Troi J Pediongco; Siddharth Jayaraman; Jennifer A Juno; Bronwyn S Meehan; Phoebe M Dewar; Emmanuel A Maze; Eduard O Roos; Basu Paudyal; Jeffrey Yw Mak; Ligong Liu; David P Fairlie; Huimeng Wang; Alexandra J Corbett; James McCluskey; Lindert Benedictus; Elma Tchilian; Paul Klenerman; Sidonia Bg Eckle

doi:10.1016/j.jbc.2024.107338

MAIT cell-MR1 reactivity is highly conserved across multiple divergent species

Matthew D Edmans, Timothy K Connelley, Sophie Morgan, Troi J Pediongco, Siddharth Jayaraman, Jennifer A Juno, Bronwyn S Meehan, Phoebe M Dewar, Emmanuel A Maze, Eduard O Roos, Basu Paudyal, Jeffrey Yw Mak, Ligong Liu, David P Fairlie, Huimeng Wang, Alexandra J Corbett, James McCluskey, Lindert Benedictus, Elma Tchilian, Paul KlenermanSidonia Bg Eckle

Research output: Contribution to journal › Article › Academic › peer-review

Abstract

Mucosal-associated invariant T (MAIT) cells are a subset of unconventional T cells that recognize small molecule metabolites presented by major histocompatibility complex class I related protein 1 (MR1), via an αβ T cell receptor (TCR). MAIT TCRs feature an essentially invariant TCR α-chain, which is highly conserved between mammals. Similarly, MR1 is the most highly conserved major histocompatibility complex-I–like molecule. This extreme conservation, including the mode of interaction between the MAIT TCR and MR1, has been shown to allow for species-mismatched reactivities unique in T cell biology, thereby allowing the use of selected species-mismatched MR1–antigen (MR1–Ag) tetramers in comparative immunology studies. However, the pattern of cross-reactivity of species-mismatched MR1–Ag tetramers in identifying MAIT cells in diverse species has not been formally assessed. We developed novel cattle and pig MR1–Ag tetramers and utilized these alongside previously developed human, mouse, and pig-tailed macaque MR1–Ag tetramers to characterize cross-species tetramer reactivities. MR1–Ag tetramers from each species identified T cell populations in distantly related species with specificity that was comparable to species-matched MR1–Ag tetramers. However, there were subtle differences in staining characteristics with practical implications for the accurate identification of MAIT cells. Pig MR1 is sufficiently conserved across species that pig MR1–Ag tetramers identified MAIT cells from the other species. However, MAIT cells in pigs were at the limits of phenotypic detection. In the absence of sheep MR1–Ag tetramers, a MAIT cell population in sheep blood was identified phenotypically, utilizing species-mismatched MR1–Ag tetramers. Collectively, our results validate the use and define the limitations of species-mismatched MR1–Ag tetramers in comparative immunology studies.

Original language	English
Article number	107338
Journal	Journal of Biological Chemistry
Volume	300
Issue number	6
Early online date	3 May 2024
DOIs	https://doi.org/10.1016/j.jbc.2024.107338
Publication status	Published - Jun 2024

Bibliographical note

Publisher Copyright:
© 2024 The Authors

Funding

We thank Dr Carl-Philipp Hackstein and Dr Chris Willberg (University of Oxford) for intellectual advice and technical assistance. We thank Dr Ted Hansen (University of Washington, Seattle, WA) and Dr Wei-Jen Chua Yankelevich (U.S. Food and Drug Administration, Washington, D.C.) for provision of hybridomas producing monoclonal antibodies 26.5 and 8F2.F9. We acknowledge the Melbourne Cytometry Platform (Peter Doherty Institute node) for the provision of flow cytometry services. We also thank the Pirbright flow cytometry facility (National Capability Science Services) for support with flow cytometry services and the Roslin Institute Veterinary Immunological Toolbox facility for monoclonal antibody production. M. D. E. L. B. E. T. P. K. and S. B. G. E. conceptualization; M. D. E. and S. B. G. E. formal analysis; M. D. E. T. K. C. S. M. T. J. P. S. J. J. A. J. B. S. M. P. M. D. E. A. M. E. O. R. B. P. and S. B. G. E. investigation; M. D. E. T. K. C. S. M. T. J. P. S. J. J. A. J. B. S. M. P. M. D. E. A. M. E. O. R. B. P. H. W. A. J. C. L. B. E. T. P. K. and S. B. G. E. methodology; M. D. E. L. B. P. K. E. T. and S. B. G. E. project administration; M. D. E. and S. B. G. E. visualization; M. D. E. and S. B. G. E. writing\u2013original draft; M. D. E. T. K. C. J. A. J. E. A. M. J. Y. W. M. D. P. F. H. W. A. J. C. J. M. L. B. E. T. P. K. and S. B. G. E. writing\u2013review and editing; J. A. J. D. P. F. A. J. C. J. M. E. T. P. K. and S. B. G. E. funding acquisition; J. Y. W. M. L. L. and D. P. F. resources; L. B. E. T. P. K. and S. B. G. E. supervision. We would like to acknowledge the following funding: Biomedical Junior Research Fellow Linacre College University of Oxford (M. D. E.), National Health and Medical Research Council (NHMRC) of Australia Investigator Grants (J. A. J.: 2009308, D. P. F.: 2009551, A. J. C.: 1193745, J. M.: 2008616, S. B. G. E.: 1196881), Dame Kate Campbell Research Fellowships from The University of Melbourne (J. A. J. A. J. C. S. B. G. E.), Australian Research Council (ARC) Center of Excellence (D. P. F.: CE200100012), National Institutes of Health RO1 (J. M. D. P. F.: AI148407-01A1), Wellcome trust investigator award (P. K.: 222426/Z/21/Z) and CRUK award (DRCNPG-Nov22/100005), Strategic Programme and Core Capability Grants to The Pirbright Institute (E. T.: BBS/E/I/00007031, BBS/E/I/00007037, and BBS/E/I/00007039) and The Roslin Institute (BBS/E/D/20002174). The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. We would like to acknowledge the following funding: National Health and Medical Research Council (NHMRC) of Australia Investigator Grants (J.A. Juno: 2009308, D.P. Fairlie: 2009551, A.J. Corbett: 1193745, J. McCluskey: 2008616, S.B.G. Eckle: 1196881), Dame Kate Campbell Research Fellowships from The University of Melbourne (J. Juno, A.J. Corbett, S. B. G. Eckle), Australian Research Council (ARC) Center of Excellence (D.P. Fairlie: CE200100012), National Institutes of Health RO1 (J. McCluskey, D.P. Fairlie: AI148407-01A1), Strategic Programme and Core Capability Grants to The Pirbright Institute (E. Tchilian: BBS/E/I/00007031, BBS/E/I/00007037 and BBS/E/I/00007039).

Funders	Funder number
Dr Wei-Jen Chua Yankelevich
Center of Excellence
University of Melbourne
University of Washington
Wellcome Trust	222426/Z/21/Z
Wellcome Trust
Australian Research Council	CE200100012
Australian Research Council
National Health and Medical Research Council	1193745, 2009551, 2008616, 2009308, 1196881
National Health and Medical Research Council
Cancer Research UK	DRCNPG-Nov22/100005
Cancer Research UK
U.S. Food and Drug Administration	8F2
U.S. Food and Drug Administration
Roslin Institute	BBS/E/D/20002174
National Institutes of Health	AI148407-01A1
National Institutes of Health
Pirbright Institute	BBS/E/I/00007037, BBS/E/I/00007039, BBS/E/I/00007031
Pirbright Institute

Keywords

5-(2-oxopropylideneamino)-6-D-ribitylaminouracil (5-OP-RU)
Antigen (Ag)
Comparative immunology
Innate-like immunity
MHC-I related protein 1 (MR1)
Major Histocompatibility Complex (MHC)
Mucosal-associated invariant T (MAIT) cell
T cell biology
T cell receptor (TCR)

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PIIS0021925824018398Final published version, 3.72 MBLicence: CC BY

Cite this

Edmans, M. D., Connelley, T. K., Morgan, S., Pediongco, T. J., Jayaraman, S., Juno, J. A., Meehan, B. S., Dewar, P. M., Maze, E. A., Roos, E. O., Paudyal, B., Mak, J. Y., Liu, L., Fairlie, D. P., Wang, H., Corbett, A. J., McCluskey, J., Benedictus, L., Tchilian, E., ... Eckle, S. B. (2024). MAIT cell-MR1 reactivity is highly conserved across multiple divergent species. Journal of Biological Chemistry, 300(6), Article 107338. https://doi.org/10.1016/j.jbc.2024.107338

@article{f928d688f89d4dcb973097d74e70d44a,

title = "MAIT cell-MR1 reactivity is highly conserved across multiple divergent species",

abstract = "Mucosal-associated invariant T (MAIT) cells are a subset of unconventional T cells that recognize small molecule metabolites presented by major histocompatibility complex class I related protein 1 (MR1), via an αβ T cell receptor (TCR). MAIT TCRs feature an essentially invariant TCR α-chain, which is highly conserved between mammals. Similarly, MR1 is the most highly conserved major histocompatibility complex-I–like molecule. This extreme conservation, including the mode of interaction between the MAIT TCR and MR1, has been shown to allow for species-mismatched reactivities unique in T cell biology, thereby allowing the use of selected species-mismatched MR1–antigen (MR1–Ag) tetramers in comparative immunology studies. However, the pattern of cross-reactivity of species-mismatched MR1–Ag tetramers in identifying MAIT cells in diverse species has not been formally assessed. We developed novel cattle and pig MR1–Ag tetramers and utilized these alongside previously developed human, mouse, and pig-tailed macaque MR1–Ag tetramers to characterize cross-species tetramer reactivities. MR1–Ag tetramers from each species identified T cell populations in distantly related species with specificity that was comparable to species-matched MR1–Ag tetramers. However, there were subtle differences in staining characteristics with practical implications for the accurate identification of MAIT cells. Pig MR1 is sufficiently conserved across species that pig MR1–Ag tetramers identified MAIT cells from the other species. However, MAIT cells in pigs were at the limits of phenotypic detection. In the absence of sheep MR1–Ag tetramers, a MAIT cell population in sheep blood was identified phenotypically, utilizing species-mismatched MR1–Ag tetramers. Collectively, our results validate the use and define the limitations of species-mismatched MR1–Ag tetramers in comparative immunology studies.",

keywords = "5-(2-oxopropylideneamino)-6-D-ribitylaminouracil (5-OP-RU), Antigen (Ag), Comparative immunology, Innate-like immunity, MHC-I related protein 1 (MR1), Major Histocompatibility Complex (MHC), Mucosal-associated invariant T (MAIT) cell, T cell biology, T cell receptor (TCR)",

author = "Edmans, {Matthew D} and Connelley, {Timothy K} and Sophie Morgan and Pediongco, {Troi J} and Siddharth Jayaraman and Juno, {Jennifer A} and Meehan, {Bronwyn S} and Dewar, {Phoebe M} and Maze, {Emmanuel A} and Roos, {Eduard O} and Basu Paudyal and Mak, {Jeffrey Yw} and Ligong Liu and Fairlie, {David P} and Huimeng Wang and Corbett, {Alexandra J} and James McCluskey and Lindert Benedictus and Elma Tchilian and Paul Klenerman and Eckle, {Sidonia Bg}",

note = "Publisher Copyright: {\textcopyright} 2024 The Authors",

year = "2024",

month = jun,

doi = "10.1016/j.jbc.2024.107338",

language = "English",

volume = "300",

journal = "Journal of Biological Chemistry",

issn = "0021-9258",

publisher = "American Society for Biochemistry and Molecular Biology Inc.",

number = "6",

}

Edmans, MD, Connelley, TK, Morgan, S, Pediongco, TJ, Jayaraman, S, Juno, JA, Meehan, BS, Dewar, PM, Maze, EA, Roos, EO, Paudyal, B, Mak, JY, Liu, L, Fairlie, DP, Wang, H, Corbett, AJ, McCluskey, J, Benedictus, L, Tchilian, E, Klenerman, P & Eckle, SB 2024, 'MAIT cell-MR1 reactivity is highly conserved across multiple divergent species', Journal of Biological Chemistry, vol. 300, no. 6, 107338. https://doi.org/10.1016/j.jbc.2024.107338

TY - JOUR

T1 - MAIT cell-MR1 reactivity is highly conserved across multiple divergent species

AU - Edmans, Matthew D

AU - Connelley, Timothy K

AU - Morgan, Sophie

AU - Pediongco, Troi J

AU - Jayaraman, Siddharth

AU - Juno, Jennifer A

AU - Meehan, Bronwyn S

AU - Dewar, Phoebe M

AU - Maze, Emmanuel A

AU - Roos, Eduard O

AU - Paudyal, Basu

AU - Mak, Jeffrey Yw

AU - Liu, Ligong

AU - Fairlie, David P

AU - Wang, Huimeng

AU - Corbett, Alexandra J

AU - McCluskey, James

AU - Benedictus, Lindert

AU - Tchilian, Elma

AU - Klenerman, Paul

AU - Eckle, Sidonia Bg

PY - 2024/6

Y1 - 2024/6

N2 - Mucosal-associated invariant T (MAIT) cells are a subset of unconventional T cells that recognize small molecule metabolites presented by major histocompatibility complex class I related protein 1 (MR1), via an αβ T cell receptor (TCR). MAIT TCRs feature an essentially invariant TCR α-chain, which is highly conserved between mammals. Similarly, MR1 is the most highly conserved major histocompatibility complex-I–like molecule. This extreme conservation, including the mode of interaction between the MAIT TCR and MR1, has been shown to allow for species-mismatched reactivities unique in T cell biology, thereby allowing the use of selected species-mismatched MR1–antigen (MR1–Ag) tetramers in comparative immunology studies. However, the pattern of cross-reactivity of species-mismatched MR1–Ag tetramers in identifying MAIT cells in diverse species has not been formally assessed. We developed novel cattle and pig MR1–Ag tetramers and utilized these alongside previously developed human, mouse, and pig-tailed macaque MR1–Ag tetramers to characterize cross-species tetramer reactivities. MR1–Ag tetramers from each species identified T cell populations in distantly related species with specificity that was comparable to species-matched MR1–Ag tetramers. However, there were subtle differences in staining characteristics with practical implications for the accurate identification of MAIT cells. Pig MR1 is sufficiently conserved across species that pig MR1–Ag tetramers identified MAIT cells from the other species. However, MAIT cells in pigs were at the limits of phenotypic detection. In the absence of sheep MR1–Ag tetramers, a MAIT cell population in sheep blood was identified phenotypically, utilizing species-mismatched MR1–Ag tetramers. Collectively, our results validate the use and define the limitations of species-mismatched MR1–Ag tetramers in comparative immunology studies.

AB - Mucosal-associated invariant T (MAIT) cells are a subset of unconventional T cells that recognize small molecule metabolites presented by major histocompatibility complex class I related protein 1 (MR1), via an αβ T cell receptor (TCR). MAIT TCRs feature an essentially invariant TCR α-chain, which is highly conserved between mammals. Similarly, MR1 is the most highly conserved major histocompatibility complex-I–like molecule. This extreme conservation, including the mode of interaction between the MAIT TCR and MR1, has been shown to allow for species-mismatched reactivities unique in T cell biology, thereby allowing the use of selected species-mismatched MR1–antigen (MR1–Ag) tetramers in comparative immunology studies. However, the pattern of cross-reactivity of species-mismatched MR1–Ag tetramers in identifying MAIT cells in diverse species has not been formally assessed. We developed novel cattle and pig MR1–Ag tetramers and utilized these alongside previously developed human, mouse, and pig-tailed macaque MR1–Ag tetramers to characterize cross-species tetramer reactivities. MR1–Ag tetramers from each species identified T cell populations in distantly related species with specificity that was comparable to species-matched MR1–Ag tetramers. However, there were subtle differences in staining characteristics with practical implications for the accurate identification of MAIT cells. Pig MR1 is sufficiently conserved across species that pig MR1–Ag tetramers identified MAIT cells from the other species. However, MAIT cells in pigs were at the limits of phenotypic detection. In the absence of sheep MR1–Ag tetramers, a MAIT cell population in sheep blood was identified phenotypically, utilizing species-mismatched MR1–Ag tetramers. Collectively, our results validate the use and define the limitations of species-mismatched MR1–Ag tetramers in comparative immunology studies.

KW - 5-(2-oxopropylideneamino)-6-D-ribitylaminouracil (5-OP-RU)

KW - Antigen (Ag)

KW - Comparative immunology

KW - Innate-like immunity

KW - MHC-I related protein 1 (MR1)

KW - Major Histocompatibility Complex (MHC)

KW - Mucosal-associated invariant T (MAIT) cell

KW - T cell biology

KW - T cell receptor (TCR)

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U2 - 10.1016/j.jbc.2024.107338

DO - 10.1016/j.jbc.2024.107338

M3 - Article

C2 - 38705391

SN - 0021-9258

VL - 300

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

IS - 6

M1 - 107338

ER -

MAIT cell-MR1 reactivity is highly conserved across multiple divergent species

Abstract

Bibliographical note

Funding

Keywords

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