Identification of cow milk epitopes to characterize and quantify disease-specific T cells in allergic children
Identification of cow milk epitopes to characterize and quantify disease-specific T cells in allergic children
Background: cow milk (CM) allergy is the most prevalent food allergy in young children in the United States and Great Britain. Current diagnostic tests are either unreliable (IgE test and skin prick test) or resource-intensive with risks (food challenges). Objective: We sought to determine whether allergen-specific T cells in CM-allergic (CMA) patients have a distinct quality and/or quantity that could potentially be used as a diagnostic marker.
Methods: using PBMCs from 147 food-allergic pediatric subjects, we mapped T-cell responses to a set of reactive epitopes in CM that we compiled in a peptide pool. This pool induced cytokine responses in in vitro cultured cells distinguishing subjects with CMA from subjects without CMA. We further used the pool to isolate and characterize antigen-specific CD4 memory T cells using flow cytometry and single-cell RNA/TCR sequencing assays.
Results: we detected significant changes in the transcriptional program and clonality of CM antigen-specific (CM+) T cells elicited by the pool in subjects with CMA versus subjects without CMA ex vivo. CM+ T cells from subjects with CMA had increased percentages of FOXP3 + cells over FOXP3 − cells. FOXP3 + cells are often equated with regulatory T cells that have suppressive activity, but CM+ FOXP3 + cells from subjects with CMA showed significant expression of interferon-responsive genes and dysregulated chemokine receptor expression compared with subjects without CMA, suggesting that these are not conventional regulatory T cells. The CM+ FOXP3 + cells were also more clonally expanded than the FOXP3 − population. We were further able to use surface markers (CD25, CD127, and CCR7) in combination with our peptide pool stimulation to quantify these CM+ FOXP3 + cells by a simple flow-cytometry assay. We show increased percentages of CM+ CD127 −CD25 + cells from subjects with CMA in an independent cohort, which could be used for diagnostic purposes. Looking specifically for T H2 cells normally associated with allergic diseases, we found a small population of clonally expanded CM+ cells that were significantly increased in subjects with CMA and that had high expression of T H2 cytokines and pathogenic T H2/T follicular helper markers.
Conclusions: overall, these findings suggest that there are several differences in the phenotypes of CM+ T cells with CM allergy and that the increase in CM+ FOXP3 + cells is a potential diagnostic marker of an allergic state. Such markers have promising applications in monitoring natural disease outgrowth and/or the efficacy of immunotherapy that will need to be validated in future studies.
Food allergy, T-cell epitopes, TCR repertoires, antigen-specific T cells, cow milk, cow milk allergy, food allergy diagnostics, pediatric food allergy, single-cell RNA-seq
1196-1209
Lewis, Sloan A.
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Sutherland, Aaron
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Soldevila, Ferran
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Westernberg, Luise
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Aoki, Minori
bf1ccbc5-7cfc-4889-a382-a988280d6582
Frazier, April
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Maiche, Synaida
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Erlewyn-Lajeunesse, Mich
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Arshad, Hasan
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Stephanie, Leonard
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Laubach, Susan
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Dantzer, Jennifer A.
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Wood, Robert A.
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Sette, Alessandro
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Seumois, Gregory
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Vijayanand, Pandurangan
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Peters, Bjoern
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November 2023
Lewis, Sloan A.
20bc2b35-9607-480c-906d-19f51a736e76
Sutherland, Aaron
127010d2-9081-4e5a-872d-7540a831a867
Soldevila, Ferran
c7dfc443-2892-4240-9ffa-93fb9c5b969c
Westernberg, Luise
01554b01-a66b-438e-9ba4-f89daa615668
Aoki, Minori
bf1ccbc5-7cfc-4889-a382-a988280d6582
Frazier, April
46e6176a-c8f8-4e33-8439-a789aec12ce5
Maiche, Synaida
3601648a-73c7-4442-a287-0ff78d301833
Erlewyn-Lajeunesse, Mich
e1763b6d-165b-45c5-9108-5dc8722220b9
Arshad, Hasan
917e246d-2e60-472f-8d30-94b01ef28958
Stephanie, Leonard
b81ef619-44a5-4601-822c-5c70fb908828
Laubach, Susan
65a48037-487f-44cc-9875-4f116f266fe4
Dantzer, Jennifer A.
4c1d8b8f-6f0f-4bde-afbb-4a42821cdb91
Wood, Robert A.
bad95ee5-f37f-4ee8-bb1a-002c30c41a9a
Sette, Alessandro
07751ad1-6f3c-4d6f-934a-828fa74c36f7
Seumois, Gregory
eb2b140f-e070-48bb-9f07-6dddae01e12e
Vijayanand, Pandurangan
641bbb69-b3e6-4f9f-9b71-ff53067ff853
Peters, Bjoern
c49863c8-d2c6-4db4-9d2c-72a7c0cbe117
Lewis, Sloan A., Sutherland, Aaron, Soldevila, Ferran, Westernberg, Luise, Aoki, Minori, Frazier, April, Maiche, Synaida, Erlewyn-Lajeunesse, Mich, Arshad, Hasan, Stephanie, Leonard, Laubach, Susan, Dantzer, Jennifer A., Wood, Robert A., Sette, Alessandro, Seumois, Gregory, Vijayanand, Pandurangan and Peters, Bjoern
(2023)
Identification of cow milk epitopes to characterize and quantify disease-specific T cells in allergic children.
Journal of Allergy and Clinical Immunology, 152 (5), .
(doi:10.1016/j.jaci.2023.07.020).
Abstract
Background: cow milk (CM) allergy is the most prevalent food allergy in young children in the United States and Great Britain. Current diagnostic tests are either unreliable (IgE test and skin prick test) or resource-intensive with risks (food challenges). Objective: We sought to determine whether allergen-specific T cells in CM-allergic (CMA) patients have a distinct quality and/or quantity that could potentially be used as a diagnostic marker.
Methods: using PBMCs from 147 food-allergic pediatric subjects, we mapped T-cell responses to a set of reactive epitopes in CM that we compiled in a peptide pool. This pool induced cytokine responses in in vitro cultured cells distinguishing subjects with CMA from subjects without CMA. We further used the pool to isolate and characterize antigen-specific CD4 memory T cells using flow cytometry and single-cell RNA/TCR sequencing assays.
Results: we detected significant changes in the transcriptional program and clonality of CM antigen-specific (CM+) T cells elicited by the pool in subjects with CMA versus subjects without CMA ex vivo. CM+ T cells from subjects with CMA had increased percentages of FOXP3 + cells over FOXP3 − cells. FOXP3 + cells are often equated with regulatory T cells that have suppressive activity, but CM+ FOXP3 + cells from subjects with CMA showed significant expression of interferon-responsive genes and dysregulated chemokine receptor expression compared with subjects without CMA, suggesting that these are not conventional regulatory T cells. The CM+ FOXP3 + cells were also more clonally expanded than the FOXP3 − population. We were further able to use surface markers (CD25, CD127, and CCR7) in combination with our peptide pool stimulation to quantify these CM+ FOXP3 + cells by a simple flow-cytometry assay. We show increased percentages of CM+ CD127 −CD25 + cells from subjects with CMA in an independent cohort, which could be used for diagnostic purposes. Looking specifically for T H2 cells normally associated with allergic diseases, we found a small population of clonally expanded CM+ cells that were significantly increased in subjects with CMA and that had high expression of T H2 cytokines and pathogenic T H2/T follicular helper markers.
Conclusions: overall, these findings suggest that there are several differences in the phenotypes of CM+ T cells with CM allergy and that the increase in CM+ FOXP3 + cells is a potential diagnostic marker of an allergic state. Such markers have promising applications in monitoring natural disease outgrowth and/or the efficacy of immunotherapy that will need to be validated in future studies.
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Accepted/In Press date: 31 July 2023
e-pub ahead of print date: 19 August 2023
Published date: November 2023
Additional Information:
Funding Information:
This work was supported by the National Institutes of Allergy and Infectious Diseases Division of the National Institutes of Health under award numbers U19AI135731 (to B.P., A.S. [Sette], and P.V.), T32AI125179 (to S.A.L.), and S10OD025052 (Illumina Novaseq6000).
Funding Information:
This work was supported by the National Institutes of Allergy and Infectious Diseases Division of the National Institutes of Health under award numbers U19AI135731 (to B.P., A.S. [Sette], and P.V.), T32AI125179 (to S.A.L.), and S10OD025052 (Illumina Novaseq6000).Disclosure of potential conflict of interest: R.A. Wood received research support from the National Institute of Allergy and Infectious Diseases, Aimmune, ALK, DBV, FARE, Genentech, Novartis, and Siolta. The rest of the authors declare that they have no relevant conflicts of interest.
Keywords:
Food allergy, T-cell epitopes, TCR repertoires, antigen-specific T cells, cow milk, cow milk allergy, food allergy diagnostics, pediatric food allergy, single-cell RNA-seq
Identifiers
Local EPrints ID: 483672
URI: http://eprints.soton.ac.uk/id/eprint/483672
ISSN: 0091-6749
PURE UUID: 39eed22b-ad05-418f-8eac-dbc51c20be64
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Date deposited: 03 Nov 2023 17:48
Last modified: 18 Mar 2024 03:46
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Contributors
Author:
Sloan A. Lewis
Author:
Aaron Sutherland
Author:
Ferran Soldevila
Author:
Luise Westernberg
Author:
Minori Aoki
Author:
April Frazier
Author:
Synaida Maiche
Author:
Mich Erlewyn-Lajeunesse
Author:
Leonard Stephanie
Author:
Susan Laubach
Author:
Jennifer A. Dantzer
Author:
Robert A. Wood
Author:
Alessandro Sette
Author:
Gregory Seumois
Author:
Pandurangan Vijayanand
Author:
Bjoern Peters
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