As shown in Figure 4 (a,c), stimulation of BMDCs with C. parvum sporozoite lysate and live antigen preparations had

a little effect on IL-6 production, while expression levels of IL-1β decreased. Treatment with Cp40 and Cp23 antigens significantly increased the expression of the inflammatory cytokines, IL-6 (Figure 4 (b)) and IL-1β (Figure 4 (d)). Because basal levels of IL-6 and IL-1β were detectable DAPT in vivo in untreated controls, we were able to observe a decrease in the levels of these cytokines in response to soluble sporozoite antigen, indicating some suppression. The patterns of cytokine responses observed in wild-type mice were absent in the MyD88 KO BMDCs, emphasizing the role of MyD88 in mediating these responses. In addition, no differences in TNFα expression were observed between the WT and MyD88 KO DCs post-treatment (data not shown). In addition to IL-12, dendritic

cells are an important source of IL-18. We observed modest levels of IL-18 in the conditioned media of BMDC (Figure 5). A significant induction of IL-18 was detected in response to the Cp40 antigen along with increases in responses to endotoxin (LPS) (Figure 5 (a,b)). No other significant levels of cytokine expression were detected with either live or solubilized sporozoite antigen or recombinant antigens. Lastly, whereas Th2 cytokines, namely IL-10, IL-5 and IL-4, could be detected, no differences in the expression levels between untreated and treated samples were observed (data not shown). MoDCs EPZ-6438 datasheet after 5 days of culture displayed typical rosetting with small dendrites. After the addition of antigen, enlarged

blast cells were observed. Similar to murine BMDCs, PD184352 (CI-1040) the MoDCs expressed high levels of CD83, CD86, CD40, HLA-DR and CD209. Expression of CD83, CD86, CD40, HLA-DR markers increased slightly (10–15%) following the treatment of sporozoites or antigens (except for Cp17), as shown in Figure 6. In contrast to the BMDC, the majority of untreated MoDCs expressed CD209 at day 5, and no change or a small decrease in expression was observed when incubated with either sporozoite or other C. parvum antigen (Figure 6). Human dendritic cells derived from monocytes were exposed to cryptosporidial antigens and responses are shown in Figure 7 (a). Sporozoite antigen preparations (solubilized and live) significantly (P < 0·05) induced IL-12 production from all 3 samples of human MoDCs compared to media-only controls. In particular, MoDCs from experiment #3 induced a markedly high expression of IL-12 (600 pg/mL) when reacted with live sporozoites as well as significant responses to the recombinant Cp40, Cp23 and P2 antigens. Cp40 was the only antigen that consistently induced IL-12p70 expression in all three samples by MoDCs (Figure 7 (b).