A novel splice variant of FcγRIIa: A risk factor for anaphylaxis in patients with hypogammaglobulinemia Joris van der Heijden, MSc, Judy Geissler, Edwin.

Slides:

Advertisements

Similar presentations

Douglas A. Plager, PhD, Ellen A. Weiss, Gail M. Kephart, BS, Robert M

Advertisements

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

Targeted deep sequencing identifies rare loss-of-function variants in IFNGR1 for risk of atopic dermatitis complicated by eczema herpeticum Li Gao, MD,

The human polysaccharide- and protein-specific immune response to Streptococcus pneumoniae is dependent on CD4+ T lymphocytes, CD14+ monocytes, and the.

A Turkish family with a novel mutation in the promoter region of the C1 inhibitor gene Suna Büyüköztürk, MD, Belgin Kesim Eroğlu, MD, Aslı Gelincik, MD,

Kuan-Wei Chen, PhD, Katharina Blatt, MSc, Wayne R

Defective calcium signaling and disrupted CD20–B-cell receptor dissociation in patients with common variable immunodeficiency disorders Annick A.J.M.

A novel kindred with inherited STAT2 deficiency and severe viral illness Leen Moens, PhD, Lien Van Eyck, MSc, MD, Dirk Jochmans, PhD, Tania Mitera, BSc,

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,

An IgE-associated polymorphism in STAT6 alters NF-κB binding, STAT6 promoter activity, and mRNA expression Michaela Schedel, PhD, Remo Frei, PhD, Christian.

Exploring the repertoire of IgE-binding self-antigens associated with atopic eczema Sabine Zeller, MSc, Claudio Rhyner, PhD, Norbert Meyer, MD, Peter.

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,

Douglas A. Plager, PhD, Ellen A. Weiss, Gail M. Kephart, BS, Robert M

Tissue factor–bearing exosome secretion from human mechanically stimulated bronchial epithelial cells in vitro and in vivo Jin-Ah Park, PhD, Asma S.

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

Human nuclear factor κB essential modulator mutation can result in immunodeficiency without ectodermal dysplasia Jordan S. Orange, MD, PhD, Ofer Levy,

Epigenetic mechanisms silence a disintegrin and metalloprotease 33 expression in bronchial epithelial cells Youwen Yang, PhD, Hans Michael Haitchi, MD,

A novel mutation in CD132 causes X-CID with defective T-cell activation and impaired humoral reactivity Taco W. Kuijpers, MD, PhD, Paul A. Baars, PhD,

CD4+ T cells in patients with chronic inflammatory rheumatic disorders show distinct levels of exhaustion Theresa Frenz, PhD, Elena Grabski, PhD, Daniela.

Human CD19 and CD40L deficiencies impair antibody selection and differentially affect somatic hypermutation Menno C. van Zelm, PhD, Sophinus J.W. Bartol,

IL-10 disrupts the Brd4-docking sites to inhibit LPS-induced CXCL8 and TNF-α expression in monocytes: Implications for chronic obstructive pulmonary disease

The C76R transmembrane activator and calcium modulator cyclophilin ligand interactor mutation disrupts antibody production and B-cell homeostasis in heterozygous.

Splicing defects in the CFTR gene: Minigene analysis of two mutations, G>C and A>G Gwendal Dujardin, Diane Commandeur, Catherine Le Jossic-Corcos,

Taco W. Kuijpers, MD, PhD, Paul A. Baars, PhD, Daan J

Combined DOCK8 and CLEC7A mutations causing immunodeficiency in 3 brothers with diarrhea, eczema, and infections Darrell L. Dinwiddie, PhD, Stephen F.

Assessing basophil activation by using flow cytometry and mass cytometry in blood stored 24 hours before analysis Kaori Mukai, PhD, Nicolas Gaudenzio,

Molecular cloning, expression, and characterization of a major 38-kd cochineal allergen Yoko Ohgiya, MS, Fumihiro Arakawa, MS, Hiroshi Akiyama, PhD, Yasuo.

Functional characterization of the atopy-associated gene PHF11

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

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

Recurrent anaphylaxis linked to pantoprazole

Mutation in IRF2BP2 is responsible for a familial form of common variable immunodeficiency disorder Michael D. Keller, MD, Rahul Pandey, PhD, Dong Li,

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

Hereditary angioedema: Molecular and clinical differences among European populations Matthaios Speletas, MD, PhD, Agnes Szilagyi, PhD, Fotis Psarros,

HLX1 gene variants influence the development of childhood asthma

Targeted deep sequencing identifies rare loss-of-function variants in IFNGR1 for risk of atopic dermatitis complicated by eczema herpeticum Li Gao, MD,

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

The Wiskott-Aldrich syndrome

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.

Inhibition of FcεRI-dependent mediator release and calcium flux from human mast cells by sialic acid–binding immunoglobulin-like lectin 8 engagement

Interleukin receptor-associated kinase-4 deficiency impairs Toll-like receptor–dependent innate antiviral immune responses Douglas R. McDonald, MD, PhD,

David M. Pyle, BS, Victoria S. Yang, MD, Rebecca S

Inhibition of human B-cell development into plasmablasts by histone deacetylase inhibitor valproic acid Anne-Kathrin Kienzler, MSc, Marta Rizzi, MD,

Defective B-cell memory in patients with Down syndrome

Fibronectin is a TH1-specific molecule in human subjects

Role of hyaluronan and hyaluronan-binding proteins in human asthma

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

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

Ectodermal dysplasia with immunodeficiency caused by a branch-point mutation in IKBKG/NEMO Sofie E. Jørgensen, MS, Pernille Bøttger, PhD, Emil Kofod-Olsen,

Fibronectin is a TH1-specific molecule in human subjects

Emma L. Wise, PhD, Kandace T. Bonner, BSc, Timothy J

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

Prostaglandin E2 resistance in granulocytes from patients with aspirin-exacerbated respiratory disease Tanya M. Laidlaw, MD, Anya J. Cutler, Molly S.

Kuan-Wei Chen, PhD, Katharina Blatt, MSc, Wayne R

Ian Ashmole, PhD, S. Mark Duffy, PhD, Mark L. Leyland, PhD, Valerie S

A novel hypomorphic mutation in STIM1 results in a late-onset immunodeficiency Heidi Schaballie, MD, Rémy Rodriguez, MS, Emmanuel Martin, PhD, Leen Moens,

Effect of IL-13 receptor α2 levels on the biological activity of IL-13 variant R110Q Allison-Lynn Andrews, PhD, Fabio Bucchieri, MD, Kazuhiko Arima, MD,

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

The human polysaccharide- and protein-specific immune response to Streptococcus pneumoniae is dependent on CD4+ T lymphocytes, CD14+ monocytes, and the.

Deficiency of caspase recruitment domain family, member 11 (CARD11), causes profound combined immunodeficiency in human subjects Polina Stepensky, MD,

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

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

Katie Frith, MD, Anne-Laure Joly, PhD, Cindy S. Ma, PhD, Stuart G

Douglas A. Plager, PhD, Ellen A. Weiss, Gail M. Kephart, BS, Robert M

Defective calcium signaling and disrupted CD20–B-cell receptor dissociation in patients with common variable immunodeficiency disorders Annick A.J.M.

Eosinophil and neutrophil extracellular DNA traps in human allergic asthmatic airways Ryszard Dworski, MD, PhD, Hans-Uwe Simon, MD, PhD, Aimee Hoskins,

Julie Negri, BA, S. Brandon Early, BA, John W

Priscilla A. Zetstra–van der Woude, MSc, J. Sebastiaan Vroegop, MD, H

Transforming growth factor β1 increases fibronectin deposition through integrin receptor α5β1 on human airway smooth muscle Lyn M. Moir, PhD, Janette.

Toll-like receptor heterodimer variants protect from childhood asthma

Presentation transcript:

A novel splice variant of FcγRIIa: A risk factor for anaphylaxis in patients with hypogammaglobulinemia Joris van der Heijden, MSc, Judy Geissler, Edwin van Mirre, PhD, Marcel van Deuren, MD, PhD, Jos W.M. van der Meer, MD, PhD, Abdulgabar Salama, MD, PhD, Timo K. van den Berg, PhD, Dirk Roos, PhD, Taco W. Kuijpers, MD, PhD Journal of Allergy and Clinical Immunology Volume 131, Issue 5, Pages 1408-1416.e5 (May 2013) DOI: 10.1016/j.jaci.2013.02.009 Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 Melting curves and gel electrophoresis of FcγRII isoforms in the CVID family. A, upper panels, A shift to the right in the melting curves of the patient (red lines) and her brother (blue lines) is observed for the FcγRIIa PCR product but not for the FcγRIIb2 PCR product. Duplicates of the patient and her brother are indicated by dashed lines. Melting curves of all other tested subjects are indicated in black. B, Analysis of the PCR products on agarose gel. The leftmost lanes contain markers. Lane 1, A healthy nonrelated control subject; lane 2, a nontemplate control subject; lane 3, a patient with CVID; lane 4, mother; lane 5, and brother. Journal of Allergy and Clinical Immunology 2013 131, 1408-1416.e5DOI: (10.1016/j.jaci.2013.02.009) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 Site of point mutation in the genome. A, An overview of the genomic organization of FCGR2, in which blocks represent exons and lines indicate introns. TM, Transmembrane domain. Exon 6* is hatched where it is spliced out. Retention of exon 6* is linked to the +871 point mutation in intron 5, as shown by the arrow. B, Pedigree of the index patient with CVID, including her healthy father and brother carrying the same FCGR2Ac.742+871A>G allele. Journal of Allergy and Clinical Immunology 2013 131, 1408-1416.e5DOI: (10.1016/j.jaci.2013.02.009) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 3 SDS-PAGE and Western blot of FcγRIIa and FcγRIIaexon6∗. Neutrophil lysates were obtained, as described in the Methods section, and run on a 10% (wt/vol) Bis-Tris gel. Thereafter, the proteins were blotted on a polyvinylidene difluoride membrane, which was probed with a polyclonal antibody against the cytoplasmic tail of FcγRIIa. Lane 1, Patient with CVID; lane 2, father of the patient with CVID; lane 3, healthy control subject not expressing FcγRIIaexon6∗. Journal of Allergy and Clinical Immunology 2013 131, 1408-1416.e5DOI: (10.1016/j.jaci.2013.02.009) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 4 Neutrophils expressing FcγRIIaexon6∗ respond more vigorously to dimeric IgG. Elastase release from neutrophils of subjects expressing FcγRIIa with (n = 4) or without (n = 14) exon 6*. Neutrophils were stimulated with dimeric IgG in the presence or absence of blocking Fab (IV.3, anti-FcγRII) fragments or with fMLP/cytochalasin B (CytoB). The difference in the response to dimeric IgG is statistically significant in the absence (P = .0077) and presence (P < .0001) of blocking Fab fragments in contrast to the response to fMLP/CytoB (P = .7, not significant). ∗∗P between .001 and .01; ∗∗∗P < .001. Journal of Allergy and Clinical Immunology 2013 131, 1408-1416.e5DOI: (10.1016/j.jaci.2013.02.009) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 5 Increased [Ca2+]i levels in IIA1.6 cells transfected with FcγRIIaexon6∗ after receptor cross-linking. IIA1.6 cells were transfected with either FcγRIIa (solid line and arrows) or FcγRIIaexon6∗ (dashed line and arrows), and these receptors were cross-linked. A, Representative [Ca2+]i traces from 1 experiment. Arrows indicate addition of goat anti-mouse IgG or thapsigargin (after [Ca2+]i stabilization). The experiment shown was repeated 6 times with identical results. B, Combined data from all experiments. All values are response peak levels minus baseline levels. Differences were significant in the case of FcγR cross-linking (P < .03, n = 6) but not in case of thapsigargin (P = .7 [not significant], n = 6). ∗P between .01 and .05. Journal of Allergy and Clinical Immunology 2013 131, 1408-1416.e5DOI: (10.1016/j.jaci.2013.02.009) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E1 Alignment of FCGR2A, FCGR2B, FCGR2C, and exon 6* of FCGR2A. Exon 6/6* sequences of FCGR2A (wild-type), FCGR2B, and FCGR2C and of the exon 6*–containing transcript, as indicated by “exon 6*”, are depicted in the gray box. Sequences outside the gray box represent part of intron 5/6 of the 3 genomic sequences. Nucleotide differences are indicated by asterisks. Underlined are the nucleotides that are found in FCGR2A and the exon 6*–containing transcript but not in FCGR2B and FCGR2C. SNPs are enclosed by parentheses. Journal of Allergy and Clinical Immunology 2013 131, 1408-1416.e5DOI: (10.1016/j.jaci.2013.02.009) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E2 FcγRIIa and FcγRIIaexon6∗ expression levels on neutrophils. Data are shown as the mean fluorescence intensity (MFI) of fluorescently labeled mAbs against FcγRII (AT10). No significant difference (NS) was found between neutrophils expressing FcγRIIa and FcγRIIaexon6∗ (P = .9859). Values were corrected by subtracting the background values measured with isotype mAb controls. Data are expressed as means ± SEMs of 3 experiments. Journal of Allergy and Clinical Immunology 2013 131, 1408-1416.e5DOI: (10.1016/j.jaci.2013.02.009) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E3 Pedigrees of 2 families with KD showing inheritance of FcγRIIa containing exon 6*. Pedigrees of 2 families with KD are indicated, showing autosomal inheritance of FcγRIIa containing exon 6*. Journal of Allergy and Clinical Immunology 2013 131, 1408-1416.e5DOI: (10.1016/j.jaci.2013.02.009) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E4 FcγRIIa and FcγRIIaexon6∗ expression levels on transfected IIA1.6 cells. Data are shown as the mean fluorescence intensity (MFI) of fluorescently labeled mAbs against FcγRII (AT10). No significant difference (NS) was found between cells transfected with FcγRIIa and FcγRIIaexon6∗ (P = .39). Values were corrected by subtracting the background values measured with isotype mAb controls. Data are expressed as means ± SEMs of 6 experiments. Journal of Allergy and Clinical Immunology 2013 131, 1408-1416.e5DOI: (10.1016/j.jaci.2013.02.009) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions