Presentation is loading. Please wait.

Presentation is loading. Please wait.

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

Published byἈλκμήνη Κουντουριώτης Modified over 5 years ago

Similar presentations

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

1 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 e5 (May 2013) DOI: /j.jaci Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

2 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  , e5DOI: ( /j.jaci ) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

3 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 A>G allele. Journal of Allergy and Clinical Immunology  , e5DOI: ( /j.jaci ) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

4 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  , e5DOI: ( /j.jaci ) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

5 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  , e5DOI: ( /j.jaci ) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

6 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  , e5DOI: ( /j.jaci ) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

7 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  , e5DOI: ( /j.jaci ) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

8 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  , e5DOI: ( /j.jaci ) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

9 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  , e5DOI: ( /j.jaci ) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

10 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  , e5DOI: ( /j.jaci ) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

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

Similar presentations

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

Douglas A. Plager, PhD, Ellen A. Weiss, Gail M. Kephart, BS, Robert M
Immune deficiency caused by impaired expression of nuclear factor-κB essential modifier (NEMO) because of a mutation in the 5′ untranslated region of.

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,

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.

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,

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

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.

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,

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,

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.

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.

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,

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

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.

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.

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,

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,

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,

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.

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,

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

Similar presentations

Ads by Google