Dectin-1, Dectin-2 and CARD9 are essential for macrophages to produce inflammatory cytokines in response to GCP-rCpa1 vaccine. Selected Th1-, Th2- and Th17-type cytokines including TNF-α, IL-12(p40), IL-12(p70), IL-4, IL-5, IL-13, IL-1α, IL-1β and IL-6 were assayed for bone marrow derived macrophages isolated from WT, Dectin-1−/− (ΔD1), Dectin-2−/− (ΔD2), or CARD9−/− (ΔC9) mice. BMMs were incubated with GCP-rCpa1 at a 1:50 ratio (black bars) or left unstimulated with PBS (white bars) for 24 hours and then cytokine levels were measured from supernatants. GCP-rCpa1 vaccine elicited a strong Th1 and Th17 proinflammatory cytokine response. In the absence of Dectin-1, Dectin-2 or adaptor molecule CARD9, macrophages produced reduced amount of TNF-α, IL-12 and IL-6 (Student t-test; *, P < 0.05). Studies were conducted in triplicate wells and data were the mean ± SEM.