title: WNT5A transforms intestinal CD8αα⁺ IELs into an unconventional phenotype with pro-inflammatory features.
abstract: BACKGROUND
  Intestinal intraepithelial lymphocytes that reside within the epithelium of the intestine form one of the main branches of the immune system. A majority of IELs express CD8α homodimer together with other molecules associated with immune regulation. Growing evidence points to the WNT signaling pathway as a pivotal piece in the immune balance and focuses on its direct regulation in intestinal epithelium. Therefore we decided to investigate its role in IELs' immune status determination.
  
  
  METHODS
  DSS colitis was induced in male C57BL mice. IELs were isolated from colon samples using mechanical dissociation followed by percoll gradient purification and Magnetic-activated cell sorting. Phenotype and cytokine production and condition with Wnts were analyzed by flow cytometry, real-time PCR or ELISA. Proliferation of lymphocytes were evaluated using CFSE dilution. Cell responses after WNT pathway interference were also evaluated.
  
  
  RESULTS
  Non-canonical WNT pathway elements represented by FZD5, WNT5A and NFATc1 were remarkably elevated in colitis IELs. The non-canonical WNT5A skewed them into a pro-inflammatory category as measured by inhibitory cell surface marker LAG3, LY49E, NKG2A and activated marker CD69 and FASL. Gaining of a pro-inflammatory marker was correlated with increased IFN-γ production but not TNF whilst decreased TGF-β and IL-10. Both interrupting WNT5A/PKC pathway and adding canonical WNT stimulants could curtail its immune-activating effect.
  
  
  CONCLUSIONS
  Canonical and non-canonical WNT signals act in opposing manners concerning determining CD8αα(+) IELs immune status. Targeting non-canonical WNT pathway may be promising in tackling inflammatory bowel disease.
authors.affiliation: Division of Gastroenterology and Hepatology, Shanghai Institute of Digestive Disease, Ren Ji Hospital, School of Medicine, Shanghai Jiaotong University, 145 Middle Shandong Road, Shanghai, 200001, China.
authors.forename: Di
authors.initials: D
authors.lastname: Zhao
authors.affiliation: Division of Gastroenterology and Hepatology, Shanghai Institute of Digestive Disease, Ren Ji Hospital, School of Medicine, Shanghai Jiaotong University, 145 Middle Shandong Road, Shanghai, 200001, China.
authors.forename: Antao
authors.initials: A
authors.lastname: Xu
authors.affiliation: Division of Gastroenterology and Hepatology, Shanghai Institute of Digestive Disease, Ren Ji Hospital, School of Medicine, Shanghai Jiaotong University, 145 Middle Shandong Road, Shanghai, 200001, China.
authors.forename: Zhanghan
authors.initials: Z
authors.lastname: Dai
authors.affiliation: Division of Gastroenterology and Hepatology, Shanghai Institute of Digestive Disease, Ren Ji Hospital, School of Medicine, Shanghai Jiaotong University, 145 Middle Shandong Road, Shanghai, 200001, China.
authors.forename: Jiangchen
authors.initials: J
authors.lastname: Peng
authors.affiliation: Division of Gastroenterology and Hepatology, Shanghai Institute of Digestive Disease, Ren Ji Hospital, School of Medicine, Shanghai Jiaotong University, 145 Middle Shandong Road, Shanghai, 200001, China.
authors.forename: Mingming
authors.initials: M
authors.lastname: Zhu
authors.affiliation: Division of Gastroenterology and Hepatology, Shanghai Institute of Digestive Disease, Ren Ji Hospital, School of Medicine, Shanghai Jiaotong University, 145 Middle Shandong Road, Shanghai, 200001, China.
authors.forename: Jun
authors.initials: J
authors.lastname: Shen
authors.affiliation: Division of Gastroenterology and Hepatology, Shanghai Institute of Digestive Disease, Ren Ji Hospital, School of Medicine, Shanghai Jiaotong University, 145 Middle Shandong Road, Shanghai, 200001, China.
authors.forename: Qing
authors.initials: Q
authors.lastname: Zheng
authors.affiliation: Division of Gastroenterology and Hepatology, Shanghai Institute of Digestive Disease, Ren Ji Hospital, School of Medicine, Shanghai Jiaotong University, 145 Middle Shandong Road, Shanghai, 200001, China. zhihuaran1962@yeah.net.
authors.forename: Zhihua
authors.initials: Z
authors.lastname: Ran
mesh_terms: D000818:Animals; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D002460:Cell Line; D003093:Colitis, Ulcerative / Q000276:immunology* / Q000378:metabolism; D016207:Cytokines / Q000378:metabolism; D004195:Disease Models, Animal; D006801:Humans; D007413:Intestinal Mucosa / Q000276:immunology* / Q000378:metabolism; D008297:Male; D008810:Mice, Inbred C57BL; D060449:Wnt Signaling Pathway / Q000276:immunology*
chemical_list: D016207:Cytokines