ORIGINAL ARTICLE Eye & skin Heat-shock pretreatment reduces expression and release of TSLP from keratinocytes under Th2 environment Jun-Kai Kao 1,2 , Cheng-Han Lee 1 , Ming-Sheng Lee 1 , Cheng-Sheng Hsu 1 , Long-Yen Tsao 1 , Yi-Giien Tsai 3,4,5 , Jeng-Jer Shieh 2,7,8 & Rei-Cheng Yang 1,6 1 Frontier Molecular Medical Research Center in Children, Changhua Christian Children Hospital, Changhua County, Taiwan; 2 Institute of Biomedical Sciences, National Chung Hsing University, Taichung City, Taiwan; 3 Department of Pediatrics, Changhua Christian Children Hospital, Changhua County, Taiwan; 4 School of Medicine, Kaohsiung Medical University, Kaohsiung City, Taiwan; 5 School of Medicine, Chung Shan Medical University, Taichung City, Taiwan; 6 Department of Pediatrics, Kaohsiung Medical University Hospital, Kaohsiung City, Taiwan; 7 Department of Education and Research, Taichung Veterans General Hospital, Taichung City, Taiwan; 8 Rong Hsing Research Center for Translational Medicine, National Chung Hsing University, Taichung City, Taiwan To cite this article: Kao J-K, Lee C-H, Lee M-S, Hsu C-S, Tsao L-Y, Tsai Y-G, Shieh J-J, Yang R-C. Heat-shock pretreatment reduces expression and release of TSLP from keratinocytes under Th2 environment. Pediatr Allergy Immunol 2016: 27: 62–69. Keywords atopic dermatitis; heat shock; heat-shock protein; thymic stromal lymphopoietin Correspondence Jeng-Jer Shieh, Room 116A, Research Building, Taichung Veterans General Hospital, No. 160, Sec. 3, Chung-Kang Rd., Taichung City, Taiwan 400 Tel.: +886 4 23592525 ext. 4052 Fax: +886 4 23592705 E-mail: shiehjj@vghtc.gov.tw and Rei-Cheng Yang, Department of Pediatrics, Changhua Christian Children Hospital, Changhua County, Taiwan, 135 Nanhsiao Street, Changhua, Taiwan 500 Tel.:+886 4 7238595 ext. 1902 Fax: +886 4 7285161 E-mail: 4808@cch.org.tw Accepted for publication 22 August 2015 DOI:10.1111/pai.12482 Abstract Background: Atopic dermatitis is a chronic, relapsing inflammatory disease of the skin. Current therapy is not curative, and recalcitrant disease is a big stress and challenge for parents and physicians. This study explored the potential role of heat- shock protein 70 (HSP-70) and its anti-inflammatory effects on keratinocyte under TH2 environment. Methods: Human keratinocyte cell line (HaCa T) was stimulated with IL-4, IL-13, and TNF-a to synthesize and secrete thymic stromal lymphopoietin (TSLP), an important cytokine of immunopathogenesis in atopic dermatitis. Heat shock was performed by immersing the cell-contained flash into a water bath of 45°C for 20 min. Cell viability, TSLP expression, and secretion of HaCa T cells were measured and compared. Possible regulatory mechanisms influencing the expression of TSLP, such as the STAT6 and NF-jB signal pathways, were investigated. Results: Heat-shock treatment induced intracellular HSP-70 expression in HaCa T cells without affecting cell viability. The induced expression and secretion of TSLP in HaCa T cells were suppressed by heat shock. The NF-jB signal pathway was inhibited by heat shock, leading to decreased TSLP expression and secretion. Conclusion: Heat stress-induced HSPs can significantly reduce the production and secretion of TSLP from HaCaT cells under Th2 environment. Thus, the evidence highlights the potential role of HSP-70 for atopic dermatitis in the future. Atopic dermatitis (AD) is a common inflammatory disease of the skin that often precedes asthma and allergic disorders. High prevalence rates in a number of other countries reveal that AD is a major problem in developing and developed countries (1). In Taiwan, the prevalence is 6.7% (2), with 40% of patients aged <20 years old. It occurs most often in infants (22.4%) and decreases with age, reaching 6.1% in 14-year-olds. The major features of AD include intense pruritus and chronic or relapsing dermatitis, which may cause secondary skin infection. In severe cases, AD can lead to school absenteeism, occupational disability, and emotional stress. Unfortunately, the current treatment is limited to relieving the severity of symptoms. A new direction of treatment is essential to improve the quality of life of patients and can decrease its burden on patients and families. Before AD goes into the chronic stage, its pathogenesis is predominantly Th2 immune response. The skin itself actually plays an important role in its immunopathogenesis. When the 62 Pediatric Allergy and Immunology 27 (2016) 62–69 ª 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd Pediatric Allergy and Immunology