Enhanced detection of antigen-specific T cells by a multiplexed AIM assay.

Enhanced detection of antigen-specific T cells by a multiplexed AIM assay.

Publication date: Jan 22, 2024

Broadly applicable methods to identify and characterize antigen-specific CD4 and CD8 T cells are key to immunology research, including studies of vaccine responses and immunity to infectious diseases. We developed a multiplexed activation-induced marker (AIM) assay that presents several advantages compared to single pairs of AIMs. The simultaneous measurement of four AIMs (CD69, 4-1BB, OX40, and CD40L) creates six AIM pairs that define CD4 T cell populations with partial and variable overlap. When combined in an AND/OR Boolean gating strategy for analysis, this approach enhances CD4 T cell detection compared to any single AIM pair, while CD8 T cells are dominated by CD69/4-1BB co-expression. Supervised and unsupervised clustering analyses show differential expression of the AIMs in defined T helper lineages and that multiplexing mitigates phenotypic biases. Paired and unpaired comparisons of responses to infections (HIV and cytomegalovirus [CMV]) and vaccination (severe acute respiratory syndrome coronavirus 2 [SARS-CoV-2]) validate the robustness and versatility of the method.

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Concepts Keywords
1bb 4-1BB (CD137)
Cd4 CD40L (CD154)
Cytomegalovirus CD69
Vaccination CP: Immunology
Versatility flow cytometry
OX40 (CD134)


Type Source Name
disease IDO assay
disease VO vaccine
disease MESH infectious diseases
disease MESH infections
disease VO vaccination
disease VO Severe acute respiratory syndrome coronavirus 2
disease VO Canada
disease MESH AIDS
disease VO USA
disease IDO infection
disease MESH viral infections
disease IDO cell
disease VO Viruses
disease VO frequency
disease IDO pathogen
drug DRUGBANK Methylergometrine
disease IDO blood
disease VO efficiency
disease MESH syndrome
disease VO Gag

Original Article

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