A peptide derived from the Wiskott-Aldrich syndrome (WAS) protein-interacting protein (WIP) restores WAS protein level and actin cytoskeleton reorganization.

Slides:

Advertisements

Similar presentations

Immune deficiency caused by impaired expression of nuclear factor-κB essential modifier (NEMO) because of a mutation in the 5′ untranslated region of.

Advertisements

Innate lymphoid cells contribute to allergic airway disease exacerbation by obesity Laetitia Everaere, PhD, Saliha Ait-Yahia, PhD, Olivier Molendi-Coste,

Plasmacytoid dendritic cell depletion in DOCK8 deficiency: Rescue of severe herpetic infections with IFN-α 2b therapy Sevgi Keles, MD, Haifa H. Jabara,

Combined immunodeficiency due to JAK3 mutation in a child presenting with skin granuloma Alessia Scarselli, MD, Silvia Di Cesare, BSc, Gigliola Di Matteo,

Reduced TH1/TH17 CD4 T-cell numbers are associated with impaired purified protein derivative–specific cytokine responses in patients with HIV-1 infection

Cell-specific activation profile of extracellular signal-regulated kinase 1/2, Jun N-terminal kinase, and p38 mitogen-activated protein kinases in asthmatic.

Broad spectrum of autoantibodies in patients with Wiskott-Aldrich syndrome and X- linked thrombocytopenia Elena Crestani, MD, Stefano Volpi, MD, Fabio.

B-cell reconstitution after lentiviral vector–mediated gene therapy in patients with Wiskott-Aldrich syndrome Maria Carmina Castiello, PhD, Samantha.

Regulation of GM-CSF expression by the transcription factor c-Maf

B-cell receptor cross-linking delays activation-induced cytidine deaminase induction and inhibits class-switch recombination to IgE Haifa H. Jabara,

Pentraxin 3 deletion aggravates allergic inflammation through a TH17-dominant phenotype and enhanced CD4 T-cell survival Jyoti Balhara, MSc, Lianyu Shan,

Chronic mucocutaneous candidiasis associated with an SH2 domain gain-of-function mutation that enhances STAT1 phosphorylation Ali Sobh, MD, Janet Chou,

Reduction of CRKL expression in patients with partial DiGeorge syndrome is associated with impairment of T-cell functions Mauro Giacomelli, PhD, Rajesh.

IL-4 confers resistance to IL-27–mediated suppression on CD4+ T cells by impairing signal transducer and activator of transcription 1 signaling Zhihong.

Cell-specific activation profile of extracellular signal-regulated kinase 1/2, Jun N-terminal kinase, and p38 mitogen-activated protein kinases in asthmatic.

Responsiveness to respiratory syncytial virus in neonates is mediated through thymic stromal lymphopoietin and OX40 ligand Junyan Han, PhD, Azzeddine.

Specificity protein 1 is pivotal in the skin’s antiviral response

Recurrent viral infections associated with a homozygous CORO1A mutation that disrupts oligomerization and cytoskeletal association Christina S. Yee,

Selective and direct activation of human neutrophils but not eosinophils by Toll-like receptor 8 Markus Janke, PhD, Jens Poth, Vera Wimmenauer, Thomas.

Expression of functional leukotriene B4 receptors on human airway smooth muscle cells Satoko Watanabe, BS, Akira Yamasaki, MD, PhD, Kiyoshi Hashimoto,

Flow cytometric measurement of STAT1 and STAT3 phosphorylation in CD4+ and CD8+ T cells—clinical applications in primary immunodeficiency diagnostics

Dawn C. Newcomb, PhD, Madison G

CD19 controls Toll-like receptor 9 responses in human B cells

Beatriz Garcillán, MS, Marina S

Exosomes from human macrophages and dendritic cells contain enzymes for leukotriene biosynthesis and promote granulocyte migration Julia Esser, BS, Ulf.

Functional analysis of protective IL1RL1 variants associated with asthma risk Vladimir Ramirez-Carrozzi, PhD, Amy Dressen, MS, Patrick Lupardus, PhD,

Human dendritic cell subset 4 (DC4) correlates to a subset of CD14dim/−CD16++ monocytes Federica Calzetti, BS, Nicola Tamassia, PhD, Alessandra Micheletti,

Toll-like receptor 9 suppression in plasmacytoid dendritic cells after IgE-dependent activation is mediated by autocrine TNF-α John T. Schroeder, PhD,

Transmembrane activator and CAML interactor (TACI) haploinsufficiency results in B- cell dysfunction in patients with Smith-Magenis syndrome Javier Chinen,

The Wiskott-Aldrich syndrome

Kathleen R. Bartemes, BA, Gail M. Kephart, BS, Stephanie J

T-cell receptor diversity is selectively skewed in T-cell populations of patients with Wiskott-Aldrich syndrome Junfeng Wu, MD, Dawei Liu, MS, Wenwei.

Posttranscriptional regulation of IL-13 in T cells: Role of the RNA-binding protein HuR Vincenzo Casolaro, MD, PhD, Xi Fang, MD, Brian Tancowny, MS, Jinshui.

Katherine G. MacDonald, BSc, Nicholas A. J

UV-B radiation induces the expression of antimicrobial peptides in human keratinocytes in vitro and in vivo Regine Gläser, MD, Fatemeh Navid, PhD, Winfried.

IL-2–inducible T-cell kinase modulates TH2-mediated allergic airway inflammation by suppressing IFN-γ in naive CD4+ T cells Arun K. Kannan, MS, Nisebita.

T-box 21 transcription factor is responsible for distorted TH2 differentiation in human peripheral CD4+ T cells Osamu Kaminuma, DVM, PhD, Fujiko Kitamura,

Role of hyaluronan and hyaluronan-binding proteins in human asthma

Impaired natural killer cell functions in patients with signal transducer and activator of transcription 1 (STAT1) gain-of-function mutations Giovanna.

Brent E. Palmer, PhD, Douglas G. Mack, PhD, Allison K

Elucidating the effects of disease-causing mutations on STAT3 function in autosomal- dominant hyper-IgE syndrome Simon J. Pelham, MSc, Helen C. Lenthall,

Sabine Leisten, MSc, Michiko K

Thymic stromal lymphopoietin activity is increased in nasal polyps of patients with chronic rhinosinusitis Deepti R. Nagarkar, PhD, Julie A. Poposki,

T-bet inhibits innate lymphoid cell–mediated eosinophilic airway inflammation by suppressing IL-9 production Ayako Matsuki, MD, Hiroaki Takatori, MD,

Immune deficiency caused by impaired expression of nuclear factor-κB essential modifier (NEMO) because of a mutation in the 5′ untranslated region of.

T-bet inhibits innate lymphoid cell–mediated eosinophilic airway inflammation by suppressing IL-9 production Ayako Matsuki, MD, Hiroaki Takatori, MD,

Autoimmune lymphoproliferative syndrome caused by a homozygous FasL mutation that disrupts FasL assembly Ali Sobh, MD, Elena Crestani, MD, Brittney Cangemi,

Xin-Zi Tang, PhD, James B. Jung, BS, Christopher D.C. Allen, PhD

IL-17E upregulates the expression of proinflammatory cytokines in lung fibroblasts Séverine Létuvé, PhD, Stéphane Lajoie-Kadoch, MSc, Séverine Audusseau,

Activation of human mast cells through the platelet-activating factor receptor Naoki Kajiwara, PhD, Tomomi Sasaki, BSc, Peter Bradding, DM, Glenn Cruse,

Sara Paveglio, PhD, MS, Erin Bennett, MS, Kelly L. Hawley, PhD, Adam P

Dysregulation of innate immune receptors on neutrophils in chronic granulomatous disease Dominik Hartl, MD, Natalie Lehmann, MD, Florian Hoffmann, MD,

Duy Pham, PhD, Sarita Sehra, PhD, Xin Sun, PhD, Mark H. Kaplan, PhD

Novel presentation of Omenn syndrome in association with aniridia

Autoimmune lymphoproliferative syndrome caused by homozygous FAS mutations with normal or residual protein expression Nourhen Agrebi, PhD, Lamia Sfaihi.

Jenny Seltmann, PhD, Lennart M

Expression of chemokines and chemokine receptors in lesional and nonlesional upper skin of patients with atopic dermatitis Eva Gros, MSc, Caroline Bussmann,

CCL17/thymus and activation-regulated chemokine induces calcitonin gene–related peptide in human airway epithelial cells through CCR4 Kandace Bonner,

Wasp venom immunotherapy expands a subpopulation of CD4+CD25+ forkhead box protein 3–positive regulatory T cells expressing the T-cell receptor Vβ2 and.

Regulation of IL-13 receptor α1 expression and signaling on human tonsillar B- lymphocyte subsets Oumnia Hajoui, PhD, Huaien Zheng, MD, PhD, Julie Guay,

Heterozygous N-terminal deletion of IκBα results in functional nuclear factor κB haploinsufficiency, ectodermal dysplasia, and immune deficiency Douglas.

Janet Chou, MD, Maxim Lutskiy, MD, PhD, Erdyni Tsitsikov, PhD, Luigi D

Thymic stromal lymphopoietin does not activate human basophils

IgG4 production is confined to human IL-10–producing regulatory B cells that suppress antigen-specific immune responses Willem van de Veen, MSc, Barbara.

First reported case of Omenn syndrome in a patient with reticular dysgenesis Lauren A. Henderson, MD, Francesco Frugoni, PhD, Gregory Hopkins, BS, Waleed.

Dysregulated NF-κB activation in Il1r8+/+/lpr and Il1r8−/−/lpr mice.

CCL17/thymus and activation-regulated chemokine induces calcitonin gene–related peptide in human airway epithelial cells through CCR4 Kandace Bonner,

Combined immunodeficiency in a patient with c-Rel deficiency

Invariant natural killer T cells from children with versus without food allergy exhibit differential responsiveness to milk-derived sphingomyelin Soma.

IL-2–inducible T-cell kinase modulates TH2-mediated allergic airway inflammation by suppressing IFN-γ in naive CD4+ T cells Arun K. Kannan, MS, Nisebita.

Presentation transcript:

A peptide derived from the Wiskott-Aldrich syndrome (WAS) protein-interacting protein (WIP) restores WAS protein level and actin cytoskeleton reorganization in lymphocytes from patients with WAS mutations that disrupt WIP binding Michel J. Massaad, PhD, Narayanaswamy Ramesh, PhD, Severine Le Bras, PhD, Silvia Giliani, PhD, Lucia D. Notarangelo, MD, Waleed Al-Herz, MD, Luigi D. Notarangelo, MD, Raif S. Geha, MD Journal of Allergy and Clinical Immunology Volume 127, Issue 4, Pages 998-1005.e2 (April 2011) DOI: 10.1016/j.jaci.2011.01.015 Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 nWIP interacts with and stabilizes WASP. A, Sequence and position of nWIP. B, Co-immunoprecipitation of WASP with WIP and nWIP in Jurkat T cells. Membranes were Western-blotted with antibodies to EGFP, WASP, and actin. The experiment is representative of 3 independent experiments with similar results. IP, Immunoprecipitation; WB, Western blot. C, Representative intracellular WASP staining in mouse T cells expressing EGFP, EGFP-WIP, or EGFP-nWIP. D, Histogram overlay showing the level of WASP in the gated EGFP+ populations. Bar graph shows the means ± SEMs of WASP levels from 3 independent experiments. Max, Maximum. Journal of Allergy and Clinical Immunology 2011 127, 998-1005.e2DOI: (10.1016/j.jaci.2011.01.015) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 WAS mRNA and protein levels in patient cells. A, Quantitative real-time PCR of WAS mRNA from EBV-B cells of patients with WAS/XLT and controls. Results are expressed as fold change from the average of WAS mRNA levels in controls. B, Representative Western blot of WASP and WIP in lysates of EBV-B cells from patients with WAS/XLT and 3 controls. C, Bar graph represents the means ± SEMs of WASP levels from 3 different experiments, expressed as fold change compared with the controls. Ctrls, Controls. Journal of Allergy and Clinical Immunology 2011 127, 998-1005.e2DOI: (10.1016/j.jaci.2011.01.015) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 3 Stabilization of WASP in EBV-B cells from WAS/XLT patients with the D77G and N204fs mutations. Representative WASP staining in EBV-B cells from patients D77G and N204fs expressing EGFP, EGFP-WIP, or EGFP-nWIP. Shown are the dot plots and the histogram overlay of WASP MFI in their gated EGFP+ populations. Ctrl, Control; Max, maximum. Journal of Allergy and Clinical Immunology 2011 127, 998-1005.e2DOI: (10.1016/j.jaci.2011.01.015) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 4 Average WASP increase in EBV-B cells from WAS/XLT patients. Bar graphs represent the means ± SEMs of WASP levels in the EGFP+ populations from 3 independent experiments for each patient. ND, Not detected; NS, not significant. Journal of Allergy and Clinical Immunology 2011 127, 998-1005.e2DOI: (10.1016/j.jaci.2011.01.015) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 5 Spreading of T cells from patients with mutations in WASP. A, Phalloidin staining of the actin cytoskeleton of T cells from control (Ctrl), and patients with L39P and D77G mutations (2 each) stimulated with immobilized anti-CD3. Bar = 10 μm. B, Pooled results represent the means ± SEMs of the percentage of spread cells from 3 controls and 2 patients (2 for each mutation). NS, not significant. Journal of Allergy and Clinical Immunology 2011 127, 998-1005.e2DOI: (10.1016/j.jaci.2011.01.015) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 6 Correction of T-cell spreading by expression of nWIP and WIP. A, Intracellular fluorescence for EGFP (top row), phalloidin (middle row), and merge (bottom row) of T cells from control (Ctrl), and patients with L39P and D77G mutations transduced with lentiviruses expressing EGFP, EGFP-WIP, or EGFP-nWIP, then stimulated with immobilized anti-CD3. Bar = 10 μm. B, Pooled results represent the means ± SEMs of the percentage of spread cells from 3 controls and 2 patients (2 for each mutation). Journal of Allergy and Clinical Immunology 2011 127, 998-1005.e2DOI: (10.1016/j.jaci.2011.01.015) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Expression of EGFP-WIP and EGFP-nWIP increases WASP levels in Jurkat T cells. Intracellular WASP staining was performed in Jurkat cells expressing EGFP-nWIP, EGFP-WIP, or EGFP. The gate indicates the population of cells used in the analysis. The histogram overlay shows the levels of WASP in the gated EGFP+ populations. The bar graph shows the means ± SEMs of WASP levels from 3 independent experiments. The respective samples, their MFIs, and bars are indicated using the same color as the histograms. WASP level was calculated by subtracting the background isotype staining from the samples’ MFIs and expressing it as a percentage of WASP in EGFP+ cells. Max, Maximum. Journal of Allergy and Clinical Immunology 2011 127, 998-1005.e2DOI: (10.1016/j.jaci.2011.01.015) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Efficiency of expression of EGFP, EGFP-WIP, and EGFP-nWIP in purified primary T cells. T cells purified from 3 controls (Ctrl) and 4 patients (2 siblings each) with the mutations L39P and D77G in the WH1/EVH1 domain of WASP were transduced with lentiviruses expressing EGFP, EGFP-WIP, and EGFP-nWIP. The expression of the fusion proteins was determined 24 hours posttransduction. Shown are representative histograms from 1 control and the 2 affected siblings. Journal of Allergy and Clinical Immunology 2011 127, 998-1005.e2DOI: (10.1016/j.jaci.2011.01.015) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions