TruCulture® is a whole blood collection and culture system that is designed to be used at clinical sites to investigate human immune responses under specific stimulation conditions. OptiMAP is a Multi-Analyte Profile (MAP) bead based immunoassay panel that measures 13 analytes that span the major immune responses (TH1, TH2, TH17, neutrophil, and monocyte/macrophage activation). The recall response of flu hemagglutinin (HA) antigens was examined in healthy adults vaccinated with the 2018-2019 annual flu vaccine. Whole blood was drawn into TruCulture tubes: null (media only), staphylococcal enterotoxin B (SEB), and null tubes spiked with 1.25mg each of 4 different recombinant HA proteins. All samples were incubated for 48 hours at 37oC. Supernatant were collected and analyzed using the OptiMAP panel. No inflammatory cytokines were detected in the null tubes from either the baseline or 2 weeks post-vaccination samples. There were no differences in cytokine production in the SEB stimulated samples between the baseline and 2 weeks post-flu vaccination time lines. For flu HA stimulated samples, there was a significant increase in production of GM-CSF, IFN-γ, IL-1β, IL-6, IL-12p70, IL-23, and TNF-α at 2 weeks post-flu vaccination compared to samples collected at baseline. These data demonstrate that TruCulture in conjunction with OptiMAP are useful tools for investigating antigen specific responses.