title: IL-4 confers resistance to IL-27-mediated suppression on CD4+ T cells by impairing signal transducer and activator of transcription 1 signaling.
abstract: BACKGROUND
  TH2 cells play a critical role in the pathogenesis of allergic asthma. Established TH2 cells have been shown to resist reprogramming into TH1 cells. The inherent stability of TH2 cells poses a significant barrier to treating allergic diseases.
  
  
  OBJECTIVE
  We sought to understand the mechanisms by which CD4(+) T cells from asthmatic patients resist the IL-27-mediated inhibition.
  
  
  METHODS
  We isolated and cultured CD4(+) T cells from both healthy subjects and allergic asthmatic patients to test whether IL-27 can inhibit IL-4 production by the cultured CD4(+) T cells using ELISA. Culturing conditions that resulted in resistance to IL-27 were determined by using both murine and human CD4(+) T-cell culture systems. Signal transducer and activator of transcription (STAT) 1 phosphorylation was analyzed by means of Western blotting and flow cytometry. Suppressor of cytokine signaling (Socs) mRNA expression was measured by using quantitative PCR. The small interfering RNA method was used to knockdown the expression of Socs3 mRNA.
  
  
  RESULTS
  We demonstrated that CD4(+) T cells from asthmatic patients resisted the suppression of IL-4 production mediated by IL-27. We observed that repeated exposure to TH2-inducing conditions rendered healthy human CD4(+) T cells resistant to IL-27-mediated inhibition. Using an in vitro murine culture system, we further demonstrated that repeated or higher doses of IL-4 stimulation, but not IL-2 stimulation, upregulated Socs3 mRNA expression and impaired IL-27-induced STAT1 phosphorylation. The knockdown of Socs3 mRNA expression restored IL-27-induced STAT1 phosphorylation and IL-27-mediated inhibition of IL-4 production.
  
  
  CONCLUSIONS
  Our findings demonstrate that differentiated TH2 cells can resist IL-27-induced reprogramming toward TH1 cells by downregulating STAT1 phosphorylation and likely explain why the CD4(+) T cells of asthmatic patients are resistant to IL-27-mediated inhibition.
authors: Division of Allergy and Immunology, Department of Medicine, National Jewish Health, Denver, Colo; Zhangshan Hospital, Fudan University, Shanghai, China.
authors: Zhihong
authors: Z
authors: Chen
authors: Shanze
authors: S
authors: Wang
authors: Nkiruka
authors: N
authors: Erekosima
authors: Yapeng
authors: Y
authors: Li
authors: Jessie
authors: J
authors: Hong
authors: Xiaopeng
authors: X
authors: Qi
authors: Patricia
authors: P
authors: Merkel
authors: Vijaya
authors: V
authors: Nagabhushanam
authors: Eugene
authors: E
authors: Choo
authors: Rohit
authors: R
authors: Katial
authors: Rafeul
authors: R
authors: Alam
authors: Anita
authors: A
authors: Trikha
authors: Hong Wei
authors: HW
authors: Chu
authors: Yonghua
authors: Y
authors: Zhuang
authors: Meiling
authors: M
authors: Jin
authors: Chunxue
authors: C
authors: Bai
authors: Hua
authors: H
authors: Huang
mesh_terms: D000818:Animals; D001249:Asthma / Q000276:immunology*; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology*; D002478:Cells, Cultured; D006801:Humans; D006969:Hypersensitivity, Immediate / Q000276:immunology; D015847:Interleukin-4 / Q000096:biosynthesis / Q000276:immunology; D007378:Interleukins / Q000276:immunology* / Q000494:pharmacology; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D010766:Phosphorylation; D060888:Real-Time Polymerase Chain Reaction; D050794:STAT1 Transcription Factor / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D050826:Suppressor of Cytokine Signaling Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism
chemical_list: D007378:Interleukins; C464565:MYDGF protein, human; D050794:STAT1 Transcription Factor; C494079:STAT1 protein, human; D050826:Suppressor of Cytokine Signaling Proteins; D015847:Interleukin-4
keywords: APC; Allophycocyanin; Asthma; High concentrations of T(H)2-inducing reagents; IL-27; IL-27 receptor; IL-27R; IL-4; Intermediate concentrations of T(H)2-inducing reagents; Low concentrations of T(H)2-inducing reagents; MFI; Mean fluorescence intensity; Memory T(H)2 cells; Memory non-T(H)2 cells, including T(H)1 and T(H)17 cells; Non-T(H)2(m); PE; Phosphorylated STAT; Phycoerythrin; Quantitative RT-PCR; Recombinant human; Recombinant murine; SOCS; STAT; Signal transducer and activator of transcription; Small interfering RNA; Suppressor of cytokine signaling; T(H)2 stability; T(H)2(hi); T(H)2(int); T(H)2(lo); T(H)2(m); T-bet; T-box transcription factor; pSTAT; qPCR; rh; rm; siRNA; signal transducer and activator of transcription 1 signaling; suppressor of cytokine signaling 3
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