Flow cytometric identification of T_fh13 cells in mouse and human - 04/02/21

Doi : 10.1016/j.jaci.2020.04.063

Jennifer S. Chen, AB ^a,^b,^{^{∗
These authors contributed equally to this work.}}, Jessica D.S. Grassmann, MS ^c,^{^{∗
These authors contributed equally to this work.}}, Uthaman Gowthaman, PhD ^a,^b,^{^{∗
These authors contributed equally to this work.}}, Sam J. Olyha, MSc ^a,^b, Tregony Simoneau, MD ^d,^e, M. Cecilia Berin, PhD ^f, Stephanie C. Eisenbarth, MD, PhD ^a,^b, Adam Williams, PhD ^c,^g,^⁎
^a Department of Laboratory Medicine, Yale University School of Medicine, New Haven, Conn
^b Department of Immunobiology, Yale University School of Medicine, New Haven, Conn
^c Jackson Laboratory for Genomic Medicine, Farmington, Conn
^d Department of Pediatrics, Harvard Medical School, Boston, Mass
^e Division of Respiratory Diseases, Boston Children's Hospital, Boston, Mass
^f Jaffe Food Allergy Institute/Immunology Institute, Icahn School of Medicine at Mount Sinai, New York
^g Department of Genetics and Genome Sciences, University of Connecticut Health Center, Farmington, Conn

^∗Corresponding author: Adam Williams, PhD, The Jackson Laboratory for Genomic Medicine, Department of Genetics and Genome Sciences, University of Connecticut Health Center, 10 Discovery Drive, Farmington, CT 06032.The Jackson Laboratory for Genomic MedicineDepartment of Genetics and Genome SciencesUniversity of Connecticut Health Center10 Discovery DriveFarmingtonCT06032

Abstract

Anaphylaxis is a life-threatening allergic reaction caused by cross-linking of high-affinity IgE antibodies on the surface of mast cells and basophils. Understanding the cellular mechanisms that lead to high-affinity IgE production is required to develop better therapeutics for preventing this severe reaction. A recently discovered population of T follicular helper T_fh13 cells regulates the production of high-affinity IgE in mouse models of allergy and can also be found in patients with allergies with IgE antibodies against food or aeroallergens. Here we describe optimized protocols for identifying T_fh13 cells in both mice and humans.

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Key words : T follicular helper cells, T_fh13 cells, intracellular cytokine staining, IL-4, IL-5, IL-13, allergy, IgE

Abbreviations used : BCL-6, cIMDM, CPE, CXCR, FACS, FcR, FMO, IMDM, LN, medLN, NP-OVA, PD-1, PMA, RBC, RT, TC, T_fh

Plan

Introduction

Identification of polyclonal and antigen-specific murine T_fh13 cells

Materials

Methods

Notes

Identification of polyclonal human T_fh13 cells

Materials

Methods

Notes

Identification of peanut-specific human T_fh13 cells

Materials

Methods

Notes

This work was supported by National Institutes of Health (NIH) grant T32GM007205 (to J.S.C. and S.J.O.), Gershon/Trudeau Fellowship from Immunobiology at Yale University (to U.G.), Robert E. Leet and Clara Guthrie Patterson Trust Mentored Research Award (to T.S.), NIH U24AI118644 (to M.C.B.), Food Allergy Research and Education (to S.C.E.), Ira and Diana Riklis Family Research Award in Food Allergy (to S.C.E.), NIH R01 AI136942 (to S.C.E.), NIH R01 AI108829 (to S.C.E.), NIH R21 AI135221 (to A.W.), NIH R21 AI133440 (to A.W.), and NIH R01 AI141609 (to A.W.).

Disclosure of potential conflict of interest: M.C. Berin serves on the scientific advisory board of Prota Therapeutics. The rest of the authors declare that they have no relevant conflicts of interest.

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Vol 147 - N° 2

P. 470-483 - février 2021 Retour au numéro

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Flow cytometric identification of T_fh13 cells in mouse and human - 04/02/21

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