These findings are consistent with our observations and altogether suggest that the presence of certain endogenous immunomodulators in blood can impact the in vitro function of antigen presenting cells in IGRA. Within-subject variability poses a major dilemma for the clinician who must decide how to manage patients with reversion or conversion of their IGRA results. OSI-774 EGFR/HER2 inhibitor studies on temporal association between human conditions and/or behavior and IGRA test results are needed to investigate the source of endogenous immunomodulatory signals. Elimination of variation in endogenous signals could enhance the reproducibility and accuracy of IGRA. The mechanism by which TLR agonists elicited an earlier and more robust IFN-c response from antigen-specific T cells in the whole blood IGRA is not known. The induction of inflammatory cytokines and IFN-a by poly and LPS are consistent with the known in vivo properties of TLR agonists as initiators of adaptive immune responses through stimulation of innate immunity. However, immunomodulation of QFT-GIT assay with purified IL-6, IL-12, and IFN-a, alone or in combination, was insufficient to recapitulate the effects of TLR agonists suggesting that other factors may be essential. The differential regulation of antigen presenting molecules and costimulatory molecules in antigen presenting cells stimulated with LPS and poly suggests that these TLR agonists may be enhancing T cell responses in the QFT-GIT assay through different mechanisms. The finding that Nod1 agonist Tri-DAP synergizes with TLR3 agonist poly in enhancing the IGRA response to M. tuberculosis antigens is consistent with animal studies showing a synergistic action between Nod1 and TLR agonists for priming of Th1 immune responses. The cellular origin of cytokines and the relative contribution of T cell subsets to IFN-c release remains to be determined. Analogous to vaccine strategies where PRR ligands are formulated to elicit desired protective immune responses, modulation of IGRA with PRR ligands may be an in vitro strategy to elicit responses from T cell subsets with prognostic value. In summary, we showed that in vitro immunomodulation of IGRA with TLR agonists is a novel strategy for eliciting T cell with M. tuberculosis. A deeper understanding of the cellular and molecular basis of immunomodulation of whole blood IGRA is necessary to reap their full potential. Phenolic choline esters, mainly sinapate choline esters, are the other major class of secondary metabolites in rapeseed. Sinapine, the choline ester of sinapic acid, is the predominant compound of that type, constituting 1–2% of the rapeseed meal. Although the sinapine biosynthesis pathway has been well investigated in Brassicaceae plants, the biological functions of sinapate choline esters are barely known. Sinapine was thought to be stored in Raphanus sativus seeds as a supply of choline, a compound that aids phosphatidylcholine biosynthesis in young seedlings. From a nutritional point of view, the presence of the major secondary metabolites, glucosinolates and sinapates, are unwanted because of their antinutritive properties. However, these compounds are very important for helping plants adapt to their biotic and abiotic environments, and in plants different classes of secondary metabolites play specific ecological functions. The glucosinolate-myrosinase system found in rape and other Brassicales is one of the best-explored plant chemical defense systems against herbivores.