Forkhead box protein 3 in human nasal polyp regulatory T cells is regulated by the protein suppressor of cytokine signaling 3 Feng Lan, MD, Nan Zhang,

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Presentation transcript:

Forkhead box protein 3 in human nasal polyp regulatory T cells is regulated by the protein suppressor of cytokine signaling 3 Feng Lan, MD, Nan Zhang, MD, PhD, Jie Zhang, MBBS, Olga Krysko, MD, PhD, Quanbo Zhang, MD, PhD, Junming Xian, MD, PhD, Lara Derycke, PhD, Yanyu Qi, MD, Kai Li, MD, Shixi Liu, MD, PhD, Ping Lin, PhD, Claus Bachert, MD, PhD Journal of Allergy and Clinical Immunology Volume 132, Issue 6, Pages 1314-1321.e3 (December 2013) DOI: 10.1016/j.jaci.2013.06.010 Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 SOCS3 expression in tissue from patients with CRSwNP. Representative images of SOCS3 immunohistochemistry in tissue from patients with CRSwNP and control subjects are shown. A and C, SOCS3+ cells (arrows) in control subjects. B and D, SOCS3+ stained cells (arrows) in patients with CRSwNP. Magnification ×100 (Fig 1, A and B) and ×400 (Fig 1, C and D). Journal of Allergy and Clinical Immunology 2013 132, 1314-1321.e3DOI: (10.1016/j.jaci.2013.06.010) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 Expression of SOCS3 and Foxp3 in patients with CRSwNP. A, RT-PCR showing SOCS3 expression in patients with CRSwNP and control subjects. B, Western blots for SOCS3 and Foxp3 proteins in patients with CRSwNP and control subjects. C, Quantification for SOCS3 protein in tissues from patients with CRSwNP. D, Quantification for Foxp3 protein in tissues from patients with CRSwNP. *P < .05 and **P < .01. Journal of Allergy and Clinical Immunology 2013 132, 1314-1321.e3DOI: (10.1016/j.jaci.2013.06.010) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 3 Expression of SOCS3 and Foxp3 proteins in Treg cells. A, PBMCs of healthy donors gated on living CD3+CD4+CD25+Foxp3+ cells cultured in RPMI: A-a, negative control; A-b, nonstimulated; A-c, stimulated with IL-2 and IL-15. B, CD4+CD25+Foxp3+ cells sorted from PBMCs of healthy donors and stained with anti-SOCS3 antibodies. C, The single cells of tissue from patients with CRSwNP gated on living CD3+CD4+CD25+Foxp3+SOCS3+ cells. Journal of Allergy and Clinical Immunology 2013 132, 1314-1321.e3DOI: (10.1016/j.jaci.2013.06.010) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 4 SOCS3 regulates Foxp3 expression in tissue from patients with CRSwNP. A-C, SOCS3 and Foxp3 expression in patients with CRSwNP on addition of Lipofectamine 2000 (lipo), transfection with negative control of siRNA (nc), or transfection with SOCS3-specific siRNA (siRNA). D-F, SOCS3 and Foxp3 expression on PANC-1 cells on addition of Lipofectamine 2000 (lipo), transfection with pEF-FLAG-I/m vector (vector), or transfection with pEF-FLAG-I/m SOCS3 plasmid (plasmid). *P < .05 and **P < .01. Journal of Allergy and Clinical Immunology 2013 132, 1314-1321.e3DOI: (10.1016/j.jaci.2013.06.010) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 5 Role of pSTAT3 in regulation of Foxp3. A and B, pSTAT3 expression in nasal tissue. C and D, pSTAT3 expression in patients with CRSwNP on addition of Lipofectamine 2000 (lipo), transfection with negative control of siRNA (nc), or transfection with SOCS3-specific siRNA (siRNA). E and F, pSTAT3 expression on PANC-1 cells on addition of Lipofectamine 2000 (lipo), transfection with pEF-FLAG-I/m vector (vector), or transfection with pEF-FLAG-I/m SOCS3 plasmid (plasmid). *P < .05. Journal of Allergy and Clinical Immunology 2013 132, 1314-1321.e3DOI: (10.1016/j.jaci.2013.06.010) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E1 Transfection efficiency of siRNA in nasal polyp tissue fragments. The blue fluorescence represents the nuclei of cells, and the green dots represent the FAM-labeled siRNA transfection control. A and B, Fluorescence images of nasal polyp tissue fragments without FAM-labeled siRNA. C and D, Fluorescence images of tissue fragments from patients with CRSwNP with FAM-labeled siRNA. The merged version of green and blue fluorescence indicates the siRNA intracellular location (white arrows). Magnification: ×100 (Fig E1, A and C) and ×400 (Fig E1, B and D). Journal of Allergy and Clinical Immunology 2013 132, 1314-1321.e3DOI: (10.1016/j.jaci.2013.06.010) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions