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2017 ; 12
(10
): e0186998
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Comparative analysis of activation induced marker (AIM) assays for sensitive
identification of antigen-specific CD4 T cells
#MMPMID29065175
Reiss S
; Baxter AE
; Cirelli KM
; Dan JM
; Morou A
; Daigneault A
; Brassard N
; Silvestri G
; Routy JP
; Havenar-Daughton C
; Crotty S
; Kaufmann DE
PLoS One
2017[]; 12
(10
): e0186998
PMID29065175
show ga
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.
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