JSID Abstracts / Journal of Dermatological Science 69 (2013) e1–e46 e13 P01-37 IL-17 and IL-22 induced IL-1B, IL-8, TNF-A secretion via inhibitor of nuclear factor KB kinase-A (IKKA) expression in HaCaT cell line Kyung Ho Lee 1,* , Kyung-Ah Cho 2 , Jin-Young Kim 2 , So-Youn Woo 2 1 Department of Dermatology, College of Medicine, The Catholic Uni- versity of Korea 2 Departments of Microbiology, School of Medicine, Ewha Womans University, Seoul, Korea The pathogenesis of psoriasis may involve the IL-23 and Th17- mediated immune responses. Th17 cells secret IL-17 and IL-22 which mediates dermal inflammation and acanthosis. As IKKAhas been previously identified as a primary regulator of keratinocyte differentiation and proliferation, we proposed that IL-17 and IL-22 might affect keratinocyte differentiation by changing the expres- sion of IKKA. We employed HaCaT cells maintained culture medium at a low calcium concentration (0.06 mM) and induced differenti- ation by switching high concentration (2.8 mM) media with IL-17 or IL-22, then compared IKKA expression and the cell cycle. We employed reconstituted human epidermal skin (Neoderm) and mice ears for the in vivo studies. Elevated calcium concentration induced IKKA expression and terminal differentiation with cell cycle arrest in HaCaT cell cultures. Moreover, IL-17 and IL-22 treat- ment also induced IKKA in HaCaT cells and reconstituted human epidermis. IKKA induction was also noted following the injection of IL-17 and IL-22 into mice ears. Although the induction of IKKA was accompanied by keratinocyte differentiation, IL-17 and IL-22 did not affect calcium-mediated differentiation or the cell cycle. Rather, In this study, we demonstrated that IL-17 and IL-22 induced IL-1B, IL-8, TNF-A secretion via IKKA expression; this suggests that through the induction of IKKA, IL-17 and IL-22 affected patho- genesis of inflammatory skin disease, e.g., psoriasis, by inducing inflammation via secretion of pro-inflammatory cytokines. http://dx.doi.org/10.1016/j.jdermsci.2012.11.335 P01-38 Thioredoxin Suppresses Irritant Dermatitis to Croton Oil via the Inhibition of Cytokines and Chemokines Production by Kerat- incytes Hai Tian 1,* , Atsushi Fukunaga 3 , Kumiko Taguchi 3 , Susumu Fujiwara 3 , Hiroshi Nagai 3 , Yoshiyuki Matsuo 2 , Junji Yodoi 2 , Chikako Nishigori 3 1 Redox bioscience Inc, Kyoto Jpan 2 Department of Biological Response, Institute for Virus Research, Kyoto University, Kyoto, Japan 3 Division of Dermatology, Department of Internal Related, Kobe Uni- versity Graduate School of Medicine, Kobe Thioredoxin(TRX) is a 12-kDa protein with redox-active dithiol in the active site -Cys-Gly-Pro-Cys- and has excellent anti-inflammatory effect in treating animal models of various inflammatory diseases. We have previously demonstrated that the irritant dermatitis to croton oil was attenuated in human TRX-overexpressing mice, but it remains unclear whether adminis- tration of recombinant human TRX protein has therapeutic effects on the irritant dermatitis. Here we showed that swelling and the infiltration of neutrophils were strongly inhibited by application of rhTRX in irritant dermatitis to croton oil. rhTRX distributed only in the epidermis under non-inflammatory condition, whereas it was detected in both the epidermis and the dermis but not in blood and urine for 24 h after development of the irritant dermatitis. We found that application of rhTRX suppressed the production of pro-inflammatory cytokines and chemokines including TNF-alpha, CXCL-1 in skin of the irritant dermatitis. In addition, treatment of PAM212 cells with rhTRX inhibited the production of cytokines and chemokines induced by croton oil. These findings suggest that application of thioredoxin prevents skin inflammatory responses and could be a promising option for the treatment of dermatitis. http://dx.doi.org/10.1016/j.jdermsci.2012.11.336 P01-39 Intravenous immunoglobulin treatment recovers the down- regulated levels of Th1 cytokines in the sera and skin of scleroderma patients Hideo Kudo * , Masatoshi Jinnin, Keitaro Yamane, Takamitsu Makino, Ikko Kajihara, Katsunari Makino, Noritoshi Honda, Wakana Nakayama, Satoshi Fukushima, Hironobu Ihn Department of Dermatology & Plastic Surgery, Graduate School of Medical and Pharmaceutical Sciences, Kumamoto University, Kumamoto, Japan Systemic sclerosis (SSc) is an autoimmune disease character- ized by fibrosis of the skin and internal organs. There is an urgent need to develop new therapeutic approaches against skin fibrosis. Although intravenous immunoglobulin (IVIG) may be one of the promising treatments, the mechanisms by which IVIG improves the fibrosis of SSc remain unknown. To compare the cytokine profile in the sera and skin of SSc patients before and after IVIG admin- istration, and try to clarify the mechanism of the effect of IVIG. Cytokine array revealed that the serum levels of IFN- and IL-12, representative Th1 cytokines, were increased by IVIG treatment, but not by placebo. The percentage of IFN-- and IL-12-positive cell number/CD4-positive T lymphocyte cell number was also signifi- cantly increased by IVIG in SSc skin. Furthermore, mRNA expression of IFN- and IL-12 in SSc skin tissue was significantly up-regulated after IVIG treatment. The expression of Th1 cytokine is reported to be decreased in SSc. Our study suggested IVIG recovered the sup- pressed levels of Th1 cytokines, and that the treatment improves skin fibrosis by correcting the Th1/Th2 balance. In order to facili- tate the clinical use of IVIG for SSc, it is necessary to perform a lager study in the future. http://dx.doi.org/10.1016/j.jdermsci.2012.11.337 P01-40 Analysis of T cells in hapten-induced contact dermatitis on TRPV1-deficient mice Hong-jin Li * , Fukumi Furukawa, Nobuo Kanazawa Department of Dermatology, Wakayama Medical University, Wakayama, Japan Transient receptor potential vanilloid receptor 1 (TRPV1) is a nonselective cation channel activated by capsaicin, heat and acid, and thus is considered to alert noxious signals to the body mainly with pain and neurogenic inflammation. To evaluate a role of TRPV1 in the antigen-specific allergic reaction with skin inflamma- tion, the hapten-induced contact dermatitis model was adopted for TRPV1-deficient mice.As reported at the last JSID meeting, stronger ear swelling was observed in TRPV-deficient mice than in control C57BL/6 mice at each challenge by both DNCB and PiCl applica- tions. Regarding the adoptive transfer experiments, in contrast to the previous report, the sensitization phase was finally proven to be responsible for the exaggerated DNCB-induced contact dermatitis