Comparative analysis of activation induced marker (AIM) assays for sensitive identification of antigen-specific CD4 T cells

• Type of Material: Article
• Authors: Samantha Reiss, La Jolla Institute for Allergy and Immunology; Amy E. Baxter, Université de Montréal; Kimberly M. Cirelli, La Jolla Institute for Allergy and Immunology; Jennifer M. Dan, La Jolla Institute for Allergy and Immunology; Antigoni Morou, Université de Montréal; Audrey Daigneault, Université de Montréal; Nathalie Brassard, Université de Montréal; Guido Silvestri, Emory University; Jean-Pierre Routy, McGill University; Colin Havenar-Daughton, La Jolla Institute for Allergy and Immunology; Shane Crotty, La Jolla Institute for Allergy and Immunology; Daniel E. Kaufmann, Université de Montréal
• Language: English
• Date: 2017-10-24
• Publisher: Public Library of Science
• Publication Version: Final Published Version
• Copyright Statement: © 2017 Reiss et al.
• License: Creative Commons Attribution 4.0 International
• Final Published Version (URL): http://dx.doi.org/10.1371/journal.pone.0186998
• Title of Journal or Parent Work: PLoS ONE
• ISSN: 1932-6203
• Volume: 12
• Issue: 10
• Start Page: e0186998
• End Page: e0186998
• Grant/Funding Information: A.E.B. is supported by a CIHR Post-doctoral Fellowship (no. 152536); J.-P.R. is the holder of the Louis Lowenstein Chair, McGill University; D.E.K. is supported by a FRQS Senior Research Scholar Award.; This work was supported by grants from NIAID (RO1 HL-092565 and AI-12796) and the Scripps CHAVI-ID (UM1 AI100663).; This work was funded in part by P51 RR000165/OD011132 to the Yerkes National Primate Research Center, the Emory Center for AIDS Research, NIH Grant # P30-AI-504 and the FRQS AIDS and Infectious Diseases Network.
• Keywords: FOLLICULAR HELPER-CELLS; Multidisciplinary Sciences; OPTIMIZATION; MEMORY TFH CELLS; Science & Technology - Other Topics; DIFFERENTIATION; ANTIBODY-RESPONSES; Science & Technology; BLOOD
• Research Categories: Health Sciences, Immunology; Health Sciences, Pathology

Abstract

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. The identification and study of antigen-specific CD4 T cells, both in peripheral blood and in tissues, is key for a broad range of immunological research, including vaccine responses and infectious diseases. Detection of these cells is hampered by both their rarity and their heterogeneity, in particular with regards to cytokine secretion profiles. These factors prevent the identification of the total pool of antigen-specific CD4 T cells by classical methods. We have developed assays for the highly sensitive detection of such cells by measuring the upregulation of surface activation induced markers (AIM). Here, we compare two such assays based on concurrent expression of CD69 plus CD40L (CD154) or expression of OX40 plus CD25, and we develop additional AIM assays based on OX40 plus PD-L1 or 4-1BB. We compare the relative sensitivity of these assays for detection of vaccine and natural infection-induced CD4 T cell responses and show that these assays identify distinct, but overlapping populations of antigen-specific CD4 T cells, a subpopulation of which can also be detected on the basis of cytokine synthesis. Bystander activation had minimal effect on AIM markers. However, some T regulatory cells upregulate CD25 upon antigen stimulation. We therefore validated AIM assays designed to exclude most T regulatory cells, for both human and non-human primate (NHP, Macaca mulatta) studies. Overall, through head-to-head comparisons and methodological improvements, we show that AIM assays represent a sensitive and valuable method for the detection of antigen-specific CD4 T cells.