Establishing Transcription Profile of Psoriasiform Cutaneous In Vitro using HaCaT Cells Stimulated with Combination of Cytokines.

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.

• Authors: Huaping Zheng, Linna Gu, Zhen Wang, Hong Zhou, Chen Zhang, Xiu Teng, Zhonglan Hu, Xiaoqiong Wei, Xiao Liu, Fanlian Zeng, Qixiang Zhao, Yan Hao, Yawen Hu, Xiaoyan Wang, Jing Hu, Jiadong Yu, Wenlin Wu, Yifan Zhou, Kaijun Cui, Nongyu Huang, Jiong Li
• Journal: Journal of visualized experiments : JoVE (J Vis Exp) Issue 169 (03 15 2021) ISSN: 1940-087X [Electronic] United States
• PMID: 33779603 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't, Video-Audio Media)

Chemical References

• Cytokines
• FLG protein, human
• Filaggrin Proteins
• Transcription Factors

Topics

• Cytokines (metabolism)
• Filaggrin Proteins
• HaCaT Cells (metabolism)
• Humans
• Psoriasis (genetics)
• Transcription Factors (metabolism)