Establishing Transcription Profile of Psoriasiform Cutaneous In Vitro using HaCaT Cells Stimulated with Combination of Cytokines
Psoriasis is a common chronic inflammatory skin disease mediated by innate and adaptive immune systems, characterized by abnormal proliferation and differentiation of epidermal keratinocytes and infiltration of inflammatory cells. Skin-specific keratinocytes are key participants in innate immunity,...
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creator | Zheng, Huaping Gu, Linna Wang, Zhen Zhou, Hong Zhang, Chen Teng, Xiu Hu, Zhonglan Wei, Xiaoqiong Liu, Xiao Zeng, Fanlian Zhao, Qixiang Hao, Yan Hu, Yawen Wang, Xiaoyan Hu, Jing Yu, Jiadong Wu, Wenlin Zhou, Yifan Cui, Kaijun Huang, Nongyu Li, Jiong |
description | Psoriasis is a common chronic inflammatory skin disease mediated by innate and adaptive immune systems, characterized by abnormal proliferation and differentiation of epidermal keratinocytes and infiltration of inflammatory cells. Skin-specific keratinocytes are key participants in innate immunity, responding to immune cells and environmental stimulation, thereby serving an important role in the immunopathogenesis of psoriasis. Here, we present a method for inducing psoriasiform keratinocytes inflammation at transcription level with HaCaT cell line using five proinflammatory cytokines combination (M5 combination), including IL-17A, IL-22, IL-1α, TNF-α, and oncostatin M. Results demonstrate that M5 combination induced HaCaT cells showed increased levels of antimicrobial peptides (BD2, S100A7, S100A8, and S100A9), chemokines, and cytokines (CXCL1, CXCL2, CXCL8, CCL20, IL-1β, IL-6 and, IL-18). The mRNA levels of keratinocytes differentiation markers (Keratin1, Keratin10, Filaggrin, and Loricrin) were down regulated, which was consistent with the transcriptome data derived from psoriasis-like keratinocytes. The method described here, therefore, establishes an in vitro psoriasiform cutaneous inflammation at transcription level and contributes to the research for molecular pathogenesis of psoriasis. |
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Skin-specific keratinocytes are key participants in innate immunity, responding to immune cells and environmental stimulation, thereby serving an important role in the immunopathogenesis of psoriasis. Here, we present a method for inducing psoriasiform keratinocytes inflammation at transcription level with HaCaT cell line using five proinflammatory cytokines combination (M5 combination), including IL-17A, IL-22, IL-1α, TNF-α, and oncostatin M. Results demonstrate that M5 combination induced HaCaT cells showed increased levels of antimicrobial peptides (BD2, S100A7, S100A8, and S100A9), chemokines, and cytokines (CXCL1, CXCL2, CXCL8, CCL20, IL-1β, IL-6 and, IL-18). The mRNA levels of keratinocytes differentiation markers (Keratin1, Keratin10, Filaggrin, and Loricrin) were down regulated, which was consistent with the transcriptome data derived from psoriasis-like keratinocytes. The method described here, therefore, establishes an in vitro psoriasiform cutaneous inflammation at transcription level and contributes to the research for molecular pathogenesis of psoriasis.</description><identifier>ISSN: 1940-087X</identifier><identifier>EISSN: 1940-087X</identifier><identifier>DOI: 10.3791/61537</identifier><identifier>PMID: 33779603</identifier><language>eng</language><publisher>United States</publisher><subject>Cytokines - metabolism ; HaCaT Cells - metabolism ; Humans ; Psoriasis - genetics ; Transcription Factors - metabolism</subject><ispartof>Journal of visualized experiments, 2021-03 (169)</ispartof><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c277t-c6f3b9b6305c6c7d5ecba327746085e53e8840ea453dcc84738ddcf7e86dcbf53</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,776,780,3830,27903,27904</link.rule.ids><linktorsrc>$$Uhttp://dx.doi.org/10.3791/61537$$EView_record_in_Journal_of_Visualized_Experiments$$FView_record_in_$$GJournal_of_Visualized_Experiments</linktorsrc><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/33779603$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Zheng, Huaping</creatorcontrib><creatorcontrib>Gu, Linna</creatorcontrib><creatorcontrib>Wang, Zhen</creatorcontrib><creatorcontrib>Zhou, Hong</creatorcontrib><creatorcontrib>Zhang, Chen</creatorcontrib><creatorcontrib>Teng, Xiu</creatorcontrib><creatorcontrib>Hu, Zhonglan</creatorcontrib><creatorcontrib>Wei, Xiaoqiong</creatorcontrib><creatorcontrib>Liu, Xiao</creatorcontrib><creatorcontrib>Zeng, Fanlian</creatorcontrib><creatorcontrib>Zhao, Qixiang</creatorcontrib><creatorcontrib>Hao, Yan</creatorcontrib><creatorcontrib>Hu, Yawen</creatorcontrib><creatorcontrib>Wang, Xiaoyan</creatorcontrib><creatorcontrib>Hu, Jing</creatorcontrib><creatorcontrib>Yu, Jiadong</creatorcontrib><creatorcontrib>Wu, Wenlin</creatorcontrib><creatorcontrib>Zhou, Yifan</creatorcontrib><creatorcontrib>Cui, Kaijun</creatorcontrib><creatorcontrib>Huang, Nongyu</creatorcontrib><creatorcontrib>Li, Jiong</creatorcontrib><title>Establishing Transcription Profile of Psoriasiform Cutaneous In Vitro using HaCaT Cells Stimulated with Combination of Cytokines</title><title>Journal of visualized experiments</title><addtitle>J Vis Exp</addtitle><description>Psoriasis is a common chronic inflammatory skin disease mediated by innate and adaptive immune systems, characterized by abnormal proliferation and differentiation of epidermal keratinocytes and infiltration of inflammatory cells. Skin-specific keratinocytes are key participants in innate immunity, responding to immune cells and environmental stimulation, thereby serving an important role in the immunopathogenesis of psoriasis. Here, we present a method for inducing psoriasiform keratinocytes inflammation at transcription level with HaCaT cell line using five proinflammatory cytokines combination (M5 combination), including IL-17A, IL-22, IL-1α, TNF-α, and oncostatin M. Results demonstrate that M5 combination induced HaCaT cells showed increased levels of antimicrobial peptides (BD2, S100A7, S100A8, and S100A9), chemokines, and cytokines (CXCL1, CXCL2, CXCL8, CCL20, IL-1β, IL-6 and, IL-18). The mRNA levels of keratinocytes differentiation markers (Keratin1, Keratin10, Filaggrin, and Loricrin) were down regulated, which was consistent with the transcriptome data derived from psoriasis-like keratinocytes. The method described here, therefore, establishes an in vitro psoriasiform cutaneous inflammation at transcription level and contributes to the research for molecular pathogenesis of psoriasis.</description><subject>Cytokines - metabolism</subject><subject>HaCaT Cells - metabolism</subject><subject>Humans</subject><subject>Psoriasis - genetics</subject><subject>Transcription Factors - metabolism</subject><issn>1940-087X</issn><issn>1940-087X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2021</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpNkN9LwzAQx4Mobur-BcmL4Es1XZqkfZQy3WDgwCm-lTRNXLRtZi5F9uafbvdD8ekO7nMf7r4IjWJyQ0UW3_KYUXGEhnGWkIik4vX4Xz9AZwDvhPAxYekpGlAqRMYJHaLvCQRZ1hZWtn3DSy9bUN6ug3UtXnhnbK2xM3gBzlsJ1jjf4LwLstWuAzxr8YsN3uEOtutTmcslznVdA34KtulqGXSFv2xY4dw1pW3lTtwL801wH7bVcIFOjKxBjw71HD3fT5b5NJo_Pszyu3mkxkKESHFDy6zklDDFlaiYVqWk_SjhJGWaUZ2mCdEyYbRSKk0ETatKGaFTXqnSMHqOrvfetXefnYZQNBZUf-r-lWLMCGdxRpMterVHlXcAXpti7W0j_aaISbENu9iF3XOXB2VXNrr6o37TpT9pB3uX</recordid><startdate>20210315</startdate><enddate>20210315</enddate><creator>Zheng, Huaping</creator><creator>Gu, Linna</creator><creator>Wang, Zhen</creator><creator>Zhou, Hong</creator><creator>Zhang, Chen</creator><creator>Teng, Xiu</creator><creator>Hu, Zhonglan</creator><creator>Wei, Xiaoqiong</creator><creator>Liu, Xiao</creator><creator>Zeng, Fanlian</creator><creator>Zhao, Qixiang</creator><creator>Hao, Yan</creator><creator>Hu, Yawen</creator><creator>Wang, Xiaoyan</creator><creator>Hu, Jing</creator><creator>Yu, Jiadong</creator><creator>Wu, Wenlin</creator><creator>Zhou, Yifan</creator><creator>Cui, Kaijun</creator><creator>Huang, Nongyu</creator><creator>Li, Jiong</creator><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope></search><sort><creationdate>20210315</creationdate><title>Establishing Transcription Profile of Psoriasiform Cutaneous In Vitro using HaCaT Cells Stimulated with Combination of Cytokines</title><author>Zheng, Huaping ; Gu, Linna ; Wang, Zhen ; Zhou, Hong ; Zhang, Chen ; Teng, Xiu ; Hu, Zhonglan ; Wei, Xiaoqiong ; Liu, Xiao ; Zeng, Fanlian ; Zhao, Qixiang ; Hao, Yan ; Hu, Yawen ; Wang, Xiaoyan ; Hu, Jing ; Yu, Jiadong ; Wu, Wenlin ; Zhou, Yifan ; Cui, Kaijun ; Huang, Nongyu ; Li, Jiong</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c277t-c6f3b9b6305c6c7d5ecba327746085e53e8840ea453dcc84738ddcf7e86dcbf53</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2021</creationdate><topic>Cytokines - metabolism</topic><topic>HaCaT Cells - metabolism</topic><topic>Humans</topic><topic>Psoriasis - genetics</topic><topic>Transcription Factors - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Zheng, Huaping</creatorcontrib><creatorcontrib>Gu, Linna</creatorcontrib><creatorcontrib>Wang, Zhen</creatorcontrib><creatorcontrib>Zhou, Hong</creatorcontrib><creatorcontrib>Zhang, Chen</creatorcontrib><creatorcontrib>Teng, Xiu</creatorcontrib><creatorcontrib>Hu, Zhonglan</creatorcontrib><creatorcontrib>Wei, Xiaoqiong</creatorcontrib><creatorcontrib>Liu, Xiao</creatorcontrib><creatorcontrib>Zeng, Fanlian</creatorcontrib><creatorcontrib>Zhao, Qixiang</creatorcontrib><creatorcontrib>Hao, Yan</creatorcontrib><creatorcontrib>Hu, Yawen</creatorcontrib><creatorcontrib>Wang, Xiaoyan</creatorcontrib><creatorcontrib>Hu, Jing</creatorcontrib><creatorcontrib>Yu, Jiadong</creatorcontrib><creatorcontrib>Wu, Wenlin</creatorcontrib><creatorcontrib>Zhou, Yifan</creatorcontrib><creatorcontrib>Cui, Kaijun</creatorcontrib><creatorcontrib>Huang, Nongyu</creatorcontrib><creatorcontrib>Li, Jiong</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Journal of visualized experiments</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext_linktorsrc</fulltext></delivery><addata><au>Zheng, Huaping</au><au>Gu, Linna</au><au>Wang, Zhen</au><au>Zhou, Hong</au><au>Zhang, Chen</au><au>Teng, Xiu</au><au>Hu, Zhonglan</au><au>Wei, Xiaoqiong</au><au>Liu, Xiao</au><au>Zeng, Fanlian</au><au>Zhao, Qixiang</au><au>Hao, Yan</au><au>Hu, Yawen</au><au>Wang, Xiaoyan</au><au>Hu, Jing</au><au>Yu, Jiadong</au><au>Wu, Wenlin</au><au>Zhou, Yifan</au><au>Cui, Kaijun</au><au>Huang, Nongyu</au><au>Li, Jiong</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Establishing Transcription Profile of Psoriasiform Cutaneous In Vitro using HaCaT Cells Stimulated with Combination of Cytokines</atitle><jtitle>Journal of visualized experiments</jtitle><addtitle>J Vis Exp</addtitle><date>2021-03-15</date><risdate>2021</risdate><issue>169</issue><issn>1940-087X</issn><eissn>1940-087X</eissn><abstract>Psoriasis is a common chronic inflammatory skin disease mediated by innate and adaptive immune systems, characterized by abnormal proliferation and differentiation of epidermal keratinocytes and infiltration of inflammatory cells. Skin-specific keratinocytes are key participants in innate immunity, responding to immune cells and environmental stimulation, thereby serving an important role in the immunopathogenesis of psoriasis. Here, we present a method for inducing psoriasiform keratinocytes inflammation at transcription level with HaCaT cell line using five proinflammatory cytokines combination (M5 combination), including IL-17A, IL-22, IL-1α, TNF-α, and oncostatin M. Results demonstrate that M5 combination induced HaCaT cells showed increased levels of antimicrobial peptides (BD2, S100A7, S100A8, and S100A9), chemokines, and cytokines (CXCL1, CXCL2, CXCL8, CCL20, IL-1β, IL-6 and, IL-18). The mRNA levels of keratinocytes differentiation markers (Keratin1, Keratin10, Filaggrin, and Loricrin) were down regulated, which was consistent with the transcriptome data derived from psoriasis-like keratinocytes. The method described here, therefore, establishes an in vitro psoriasiform cutaneous inflammation at transcription level and contributes to the research for molecular pathogenesis of psoriasis.</abstract><cop>United States</cop><pmid>33779603</pmid><doi>10.3791/61537</doi></addata></record> |
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subjects | Cytokines - metabolism HaCaT Cells - metabolism Humans Psoriasis - genetics Transcription Factors - metabolism |
title | Establishing Transcription Profile of Psoriasiform Cutaneous In Vitro using HaCaT Cells Stimulated with Combination of Cytokines |
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