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However, it is inconsistent with the fact that CCL3 + 19 (7 : 3) did not suppress the production of IL-12 by DCs (Fig. 8b). The suppressive effects of maturation on DCs such as the reduction of IL-12 production31,47,59,60 were probably not observed in this study. LPS-treated DCs expressed MHC Class II molecules despite the presence of CCL3 + 19 (7 : 3) (Fig. 5a,c), although they did not produce cytokines at high levels (Fig. 8b). In addition, DCs pre-treated with CCL3 + 19 (7 : 3) secreted IL-6 and CXCL8 (IL-8) at levels higher than iDCs before LPS treatment (Fig. 8c,d) but they expressed CD86 or MHC Class II molecules at levels similar or lower than iDCs before LPS treatment (Fig. 5a,b). Thus, the suppressive effects of CCL3 + 19 (7 : 3) on DC maturation seem to be limited. These results indicate that DCs pre-treated with CCL3 + 19 (7 : 3) are in a semi-maturation-like state at the time of LPS stimulation. It is unclear how they suppress subsequent LPS-induced maturation and cytokine production of DCs. One possible mechanism is that CCL3 + 19 (7 : 3) induces the secretion of soluble factors from DCs that can suppress LPS-induced cytokine production or maturation of DCs. For example, TGF-β and IDO can inhibit maturation of DCs, thus suppressing the production of cytokines such as IL-12 by activated DCs.15,63,64 Similarly, IL-10 can suppress the production of IL-12 and IL-6 by DCs, leading to a semi-maturation-like state.65

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