Ignaling promotes the migration and maturation of cutaneous DCs, thereby initiating the CHS response on exposure to antigens. On the contrary, the PGE2-EP3 signaling functions in the opposite direction, balancing the cutaneous immune homeostasis in a CP21 subtle way. When the amount of antigens does not reach the threshold for appropriate immune responses, the PGE2-EP3 signaling actively limits the migration and maturation of cutaneous DCs through Gi protein to avoid unwanted inflammation. Once the antigen dose crosses the threshold, the PGE2-EP4 signaling axis overcomes the restriction by PGE2-EP3 signaling in order to switch the mode of cutaneous DCs toward activation. We do not yet know the molecular mechanism that determines which EP subtype will dominate in response to PGE2. As DCs express both EP3 and EP4, the cell surface expression level of EP3 and EP4 may be differentially regulated depending on the antigen dose. 16574785 However, at the transcript level, the mRNA level of EP4 was at least tenfold higher than that of EP3 (see Figures 1a and 2b). Therefore, it is not easy to explain the mechanism simply by the expression level of these subtypes. Alternatively, there may be cross talk between EP3 and EP4. For example, EP3-coupling Gi may somehow over-rule EP4-coupling Gs under non-inflammatory conditions when the production of PGE2 is low. On the other hand, once DCs are exposed to a large dose of antigens, EP4coupling Gs now dominates the relation (see Figure 6). In line with the above hypothesis, the binding affinity of EP3 for PGE2 is much higher than that of EP4 [21]. EP3 is the 1315463 only prostanoid receptor that couples Gi and functions in a cAMP-inhibitory manner. Other prostanoid receptors work in an either Ca2+ stimulatory (EP1, FP, TP) or cAMP-stimulatory fashion (EP2, EP4, DP, IP). This multiplicity of EP subtypes makes PGE2 the most versatile prostaglandin in vivo. Here we revealed another unexpected dual role of PGE2 on the CHS response (see Figure 6). In the steady state, low-dose PGE2 limits migration and maturation of cutaneous DCs through EP3 to halt impetuous response to suboptimal stimuli. Thus, PGE2-EP3 axis seems to exhibit fine-tuning excessive skin inflammation by restricting DC functions. This limitation is easily cancelled under inflammatory state by highdose PGE2, which now acts on EP4 to switch the state of cutaneous DCs to an activation mode. The mechanism to initiate skin immune responses have been vigorously studied, but the mechanism how to keep skin homeostasis has not been revealed well. In this study, we focused on the role of DCs. On the hand, other possible candidates to maintain skin homeostasis include regulatory T cells (Tregs). In the absence of Tregs, mice lead to spontaneous skin inflammation [23] and enhanced CHS to hapten exposure [24?6]. It remains unclear whether PGE2EP3 signaling on DCs modulates the induction of Tregs, which will be 301353-96-8 chemical information addressed in the future. It has also been reported that PGE2-EP3 signaling suppressed conjunctivitis and airway inflammation by inhibitionEP3 Signaling Regulates the Cutaneous DC FunctionsFigure 6. Hypothesis of the dual roles of PGE2 on cutaneous DCs. In the steady state when the concentration of PGE2 is low, endogenous PGE2 binds to EP3 preferentially (binding affinity of PGE2 to EP3 is higher than EP4), resulting in the prevention of impetuous immune responses to innocuous stimuli. On the other hand, in the inflammatory state, abundant PGE2 is produced by keratinocytes.Ignaling promotes the migration and maturation of cutaneous DCs, thereby initiating the CHS response on exposure to antigens. On the contrary, the PGE2-EP3 signaling functions in the opposite direction, balancing the cutaneous immune homeostasis in a subtle way. When the amount of antigens does not reach the threshold for appropriate immune responses, the PGE2-EP3 signaling actively limits the migration and maturation of cutaneous DCs through Gi protein to avoid unwanted inflammation. Once the antigen dose crosses the threshold, the PGE2-EP4 signaling axis overcomes the restriction by PGE2-EP3 signaling in order to switch the mode of cutaneous DCs toward activation. We do not yet know the molecular mechanism that determines which EP subtype will dominate in response to PGE2. As DCs express both EP3 and EP4, the cell surface expression level of EP3 and EP4 may be differentially regulated depending on the antigen dose. 16574785 However, at the transcript level, the mRNA level of EP4 was at least tenfold higher than that of EP3 (see Figures 1a and 2b). Therefore, it is not easy to explain the mechanism simply by the expression level of these subtypes. Alternatively, there may be cross talk between EP3 and EP4. For example, EP3-coupling Gi may somehow over-rule EP4-coupling Gs under non-inflammatory conditions when the production of PGE2 is low. On the other hand, once DCs are exposed to a large dose of antigens, EP4coupling Gs now dominates the relation (see Figure 6). In line with the above hypothesis, the binding affinity of EP3 for PGE2 is much higher than that of EP4 [21]. EP3 is the 1315463 only prostanoid receptor that couples Gi and functions in a cAMP-inhibitory manner. Other prostanoid receptors work in an either Ca2+ stimulatory (EP1, FP, TP) or cAMP-stimulatory fashion (EP2, EP4, DP, IP). This multiplicity of EP subtypes makes PGE2 the most versatile prostaglandin in vivo. Here we revealed another unexpected dual role of PGE2 on the CHS response (see Figure 6). In the steady state, low-dose PGE2 limits migration and maturation of cutaneous DCs through EP3 to halt impetuous response to suboptimal stimuli. Thus, PGE2-EP3 axis seems to exhibit fine-tuning excessive skin inflammation by restricting DC functions. This limitation is easily cancelled under inflammatory state by highdose PGE2, which now acts on EP4 to switch the state of cutaneous DCs to an activation mode. The mechanism to initiate skin immune responses have been vigorously studied, but the mechanism how to keep skin homeostasis has not been revealed well. In this study, we focused on the role of DCs. On the hand, other possible candidates to maintain skin homeostasis include regulatory T cells (Tregs). In the absence of Tregs, mice lead to spontaneous skin inflammation [23] and enhanced CHS to hapten exposure [24?6]. It remains unclear whether PGE2EP3 signaling on DCs modulates the induction of Tregs, which will be addressed in the future. It has also been reported that PGE2-EP3 signaling suppressed conjunctivitis and airway inflammation by inhibitionEP3 Signaling Regulates the Cutaneous DC FunctionsFigure 6. Hypothesis of the dual roles of PGE2 on cutaneous DCs. In the steady state when the concentration of PGE2 is low, endogenous PGE2 binds to EP3 preferentially (binding affinity of PGE2 to EP3 is higher than EP4), resulting in the prevention of impetuous immune responses to innocuous stimuli. On the other hand, in the inflammatory state, abundant PGE2 is produced by keratinocytes.
