Despite their crucial role in controlling viral infections and shaping disease pathogenesis, antigen-specific T-cell responses remain understudied due to the lack of simple, reliable, and scalable assays. To address this gap, we present a novel, sensitive, and rapid whole-blood assay (WBA) that enables the evaluation of virus-specific T-cell responses using small blood volumes, without the need for PBMC isolation. The assay delivers results in under 24 hours and supports the assessment of functional T cells of any specificity, making it suitable for classical research, large clinical trials, routine diagnostics, and field studies.
We validated the WBA for the precise evaluation of virus-specific T-cell responses targeting Coronaviridae, Hepadnaviridae, Paramyxoviridae, Orthomyxoviridae, Bunyaviridae, and Herpesviridae, as well as TCR-redirected T cells, in human blood. Briefly, 320 μL of whole blood is stimulated overnight with overlapping 15-mer peptides spanning the viral protein of interest. Virus-specific T cells secrete cytokines into the plasma, which are quantified using multiplex immunoassays, enabling the assessment of response magnitude and functional polarity1.
We present snapshots of diverse WBA applications, illustrating its broad utility and advantages compared with conventional T-cell assays:
Taken together, the WBA represents a versatile platform that enables virus-specific T-cell monitoring beyond specialized laboratories. Its rapid (<24 h) readout and minimal blood requirement make it broadly applicable across diverse clinical and field settings, including those with limited technical infrastructure and specialized expertise.