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

TitleComparative analysis of activation induced marker (AIM) assays for sensitive identification of antigen-specific CD4 T cells.
Publication TypeJournal Article
Year of Publication2017
AuthorsReiss S, Baxter AE, Cirelli KM, Dan JM, Morou A, Daigneault A, Brassard N, Silvestri G, Routy J-P, Havenar-Daughton C, Crotty S, Kaufmann DE
JournalPLoS One
Volume12
Issue10
Paginatione0186998
Date Published10/24/2017
ISSN1932-6203
KeywordsAnimals, Antigens, Antigens, CD, Biomarkers, Bystander Effect, CD4-Positive T-Lymphocytes, Cohort Studies, Humans, Lymphocyte Activation, Macaca mulatta
Abstract

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.

DOI10.1371/journal.pone.0186998
Alternate JournalPLoS ONE
PubMed ID29065175
PubMed Central IDPMC5655442
Grant ListR01 AI124796 / AI / NIAID NIH HHS / United States
UM1 AI100663 / AI / NIAID NIH HHS / United States
P51 OD011132 / OD / NIH HHS / United States
P51 RR000165 / RR / NCRR NIH HHS / United States
R01 HL092565 / HL / NHLBI NIH HHS / United States
CHAVI-ID: 
1
Cover Picture: