Presentation is loading. Please wait.

Presentation is loading. Please wait.

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

Published byJean-Christophe Ducharme Modified over 5 years ago

Similar presentations

Presentation on theme: "B-cell receptor cross-linking delays activation-induced cytidine deaminase induction and inhibits class-switch recombination to IgE  Haifa H. Jabara,"— Presentation transcript:

1 B-cell receptor cross-linking delays activation-induced cytidine deaminase induction and inhibits class-switch recombination to IgE  Haifa H. Jabara, BS, Jayanta Chaudhuri, PhD, Shilpee Dutt, PhD, Fatma Dedeoglu, MD, Yu Weng, BS, Michael M. Murphy, BS, Sonia Franco, MD, PhD, Fredrick W. Alt, PhD, John Manis, MD, Raif S. Geha, MD  Journal of Allergy and Clinical Immunology  Volume 121, Issue 1, Pages e2 (January 2008) DOI: /j.jaci Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

2 Fig 1 BCR cross-linking inhibits LPS plus IL-4–induced IgG1 and IgE isotype switching by murine B cells. A, Effect of goat IgG F(ab′)2 anti-mouse IgM (α-μ; 0-10 μg/mL) and control (C) goat IgG F(ab′)2 (10 μg/mL) on IgG1 and IgE synthesis by purified splenic B cells. Results represent the mean ± SE of 5 experiments (n = 5). B, Surface IgG1 expression. Results are representative of 4 experiments; percentages of sIgG1+ B cells are indicated. Journal of Allergy and Clinical Immunology  , e2DOI: ( /j.jaci ) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

3 Fig 2 No effect of BCR cross-linking on B-cell viability or proliferation. A, Tritiated thymidine incorporation (n = 3). B, CFSE dilution on day 6. C, Annexin V staining. In Fig 2, B and C, a representative of 3 experiments is shown as overlay histograms of B cells stimulated with IL-4 plus LPS (green), IL-4 plus LPS plus α-μ (10 μg/mL; red), and IL-4 plus LPS plus control (10 μg/mL; blue). Journal of Allergy and Clinical Immunology  , e2DOI: ( /j.jaci ) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

4 Fig 3 BCR cross-linking inhibits LPS/IL-4–driven switch recombination and plasmablast generation. A, RT-PCR of Cɛ GLTs, AID, and Iμ-Cɛ. B, Sμ→Sɛ by means of digestion circularization PCR. C, Semiquantitative analysis of Iμ-Cμ and Cɛ GLT by using 4-fold dilution of cDNA. D, IgM secretion (n = 5). E, CD138 expression. β2-Microglobulin (β2 m) and nicotinic acetylcholine receptor β unit (nAChR) are loading controls. Results represent 3 experiments in Fig 3, A through C, and 2 experiments in Fig 3, E. Journal of Allergy and Clinical Immunology  , e2DOI: ( /j.jaci ) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

5 Fig 4 BCR cross-linking causes a decrease in DNA repair foci. A, Immuno-FISH with 53BP1 and IgH locus bacterial artificial chromosome (BAC) probes. Representative B cell showing colocalization (white arrow) of IgH BAC probe (green) and 53BP1 protein (red). B, Quantitative analysis of immuno-FISH assay performed on primary B cells after 72 hours of culture (n = 3). ∗∗P < .01. Journal of Allergy and Clinical Immunology  , e2DOI: ( /j.jaci ) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

6 Fig 5 BCR cross-linking delays AID mRNA expression. A, Quantitative PCR analysis of AID mRNA expression on days 2, 3, and 4 after stimulation. AID mRNA levels in α-μ or control treated cultures are expressed as a percentage of AID induced by LPS plus IL-4 (n = 4). ∗P < .05; ∗∗P < .01. B, Effect of BCR cross-linking on IgE and IgG1 production 8 days after stimulation (n = 3). Journal of Allergy and Clinical Immunology  , e2DOI: ( /j.jaci ) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

7 Fig 6 BCR cross-linking has no effect on AID. A, Western blot of AID in B-cell lysates (15 μg per lane) on day 4, with Ku70 as a control. B, Assay of B-cell extracts (5 μg) for DNA CDA, with LPS-stimulated AID−/− B cells as a negative control. C, JNK phosphorylation (0.5 × 106 cells per lane). Results represent 3 experiments for Fig 6, A, and 2 experiments for Fig 2, B and C. Journal of Allergy and Clinical Immunology  , e2DOI: ( /j.jaci ) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

8 Effect of BCR cross-linking on IgG1 and IgE isotype switching by murine B cells negatively sorted for IgG, IgA, and IgE and stimulated with LPS plus IL-4. Effect of goat IgG F(ab′)2 anti-mouse IgM (α-μ; 0-10 mg/mL) and control (C) goat IgG F(ab′)2 (10 mg/mL) on IgG1 and IgE synthesis by sorted splenic B cells. Results shown are representative of 3 independent experiments. Mouse splenic B cells were purified by means of negative sorting with biotinylated mAbs to IgG1, IgG2a, IgG2b, IgG3, IgA, IgE, CD138, CD43, CD11b, and Thy-1.2 (PharMingen) and Dynabeads M-280 streptavidin (Dynal). Journal of Allergy and Clinical Immunology  , e2DOI: ( /j.jaci ) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

9 Two-color region FISH to detect chromosomal breaks at the IgH locus
Two-color region FISH to detect chromosomal breaks at the IgH locus. A, Two-color IgH FISH strategy to detect chromosomal breaks. B, Metaphase spreads showing intact IgH locus with contiguous localization of 5′ IgH (green) and 3′ IgH (red) probes. C, Chromosomal breaks in B cells from BALB/c (wild-type) mice treated with LPS/IL-4 in the presence of medium, anti-IgM, or control IgG. Stimulated H2AX−/− B cells were used as a positive control for detection of chromosomal breaks. The 3′ end of the IgH locus was detected by using BAC199 and the 5′ end was detected by using BAC207 in metaphase spreads of B cells 96 hours after stimulation, as previously described.E1 Journal of Allergy and Clinical Immunology  , e2DOI: ( /j.jaci ) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

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

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.

Immune deficiency caused by impaired expression of nuclear factor-κB essential modifier (NEMO) because of a mutation in the 5′ untranslated region of.
Barbara Stanic, PhD, Willem van de Veen, PhD, Oliver F

Barbara Stanic, PhD, Willem van de Veen, PhD, Oliver F
Expression of IL-9 receptor α chain on human germinal center B cells modulates IgE secretion  Lama M. Fawaz, PhD, Ehssan Sharif-Askari, PhD, Oumnia Hajoui,

Expression of IL-9 receptor α chain on human germinal center B cells modulates IgE secretion  Lama M. Fawaz, PhD, Ehssan Sharif-Askari, PhD, Oumnia Hajoui,
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.
IgE cross-linking impairs monocyte antiviral responses and inhibits influenza-driven TH1 differentiation  Regina K. Rowe, MD, PhD, David M. Pyle, MD,

IgE cross-linking impairs monocyte antiviral responses and inhibits influenza-driven TH1 differentiation  Regina K. Rowe, MD, PhD, David M. Pyle, MD,
Umasundari Sivaprasad, PhD, David J. Askew, PhD, Mark B

Umasundari Sivaprasad, PhD, David J. Askew, PhD, Mark B
Signal transduction pathways triggered by the FcϵRIIb receptor (CD23) in human monocytes lead to nuclear factor-κB activation  Rosa M. Ten, MD, PhDa,

Signal transduction pathways triggered by the FcϵRIIb receptor (CD23) in human monocytes lead to nuclear factor-κB activation  Rosa M. Ten, MD, PhDa,
Toll-like receptor 7–induced naive human B-cell differentiation and immunoglobulin production  Mark C. Glaum, MD, PhD, Shilpi Narula, MD, Decheng Song,

Toll-like receptor 7–induced naive human B-cell differentiation and immunoglobulin production  Mark C. Glaum, MD, PhD, Shilpi Narula, MD, Decheng Song,
IL-10 disrupts the Brd4-docking sites to inhibit LPS-induced CXCL8 and TNF-α expression in monocytes: Implications for chronic obstructive pulmonary disease 

IL-10 disrupts the Brd4-docking sites to inhibit LPS-induced CXCL8 and TNF-α expression in monocytes: Implications for chronic obstructive pulmonary disease 
Ting-ting Zhang, MSc, Klaus Okkenhaug, PhD, Baher F

Ting-ting Zhang, MSc, Klaus Okkenhaug, PhD, Baher F
Insulin-like growth factor 2 enhances regulatory T-cell functions and suppresses food allergy in an experimental model  Gui Yang, MD, Xiao-Rui Geng, MD,

Insulin-like growth factor 2 enhances regulatory T-cell functions and suppresses food allergy in an experimental model  Gui Yang, MD, Xiao-Rui Geng, MD,
Epicutaneous sensitization results in IgE-dependent intestinal mast cell expansion and food-induced anaphylaxis  Lisa M. Bartnikas, MD, Michael F. Gurish,

Epicutaneous sensitization results in IgE-dependent intestinal mast cell expansion and food-induced anaphylaxis  Lisa M. Bartnikas, MD, Michael F. Gurish,
CD40-mediated p38 mitogen-activated protein kinase activation is required for immunoglobulin class switch recombination to IgE  Ke Zhang, MD, PhD, Ling.

CD40-mediated p38 mitogen-activated protein kinase activation is required for immunoglobulin class switch recombination to IgE  Ke Zhang, MD, PhD, Ling.
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.
Efficient cytokine-induced IL-13 production by mast cells requires both IL-33 and IL-3  Ilkka S. Junttila, MD, PhD, Cynthia Watson, BSc, Laura Kummola,

Efficient cytokine-induced IL-13 production by mast cells requires both IL-33 and IL-3  Ilkka S. Junttila, MD, PhD, Cynthia Watson, BSc, Laura Kummola,
Selective and direct activation of human neutrophils but not eosinophils by Toll-like receptor 8  Markus Janke, PhD, Jens Poth, Vera Wimmenauer, Thomas.

Selective and direct activation of human neutrophils but not eosinophils by Toll-like receptor 8  Markus Janke, PhD, Jens Poth, Vera Wimmenauer, Thomas.
Surfactant protein D inhibits TNF-α production by macrophages and dendritic cells in mice  László Hortobágyi, MS, Sonja Kierstein, PhD, Kateryna Krytska,

Surfactant protein D inhibits TNF-α production by macrophages and dendritic cells in mice  László Hortobágyi, MS, Sonja Kierstein, PhD, Kateryna Krytska,
Dawn C. Newcomb, PhD, Madison G

Dawn C. Newcomb, PhD, Madison G
CD19 controls Toll-like receptor 9 responses in human B cells

CD19 controls Toll-like receptor 9 responses in human B cells
Forkhead box protein 3 demethylation is associated with tolerance induction in peanut- induced intestinal allergy  Meiqin Wang, MD, PhD, Ivana V. Yang,

Forkhead box protein 3 demethylation is associated with tolerance induction in peanut- induced intestinal allergy  Meiqin Wang, MD, PhD, Ivana V. Yang,

Similar presentations

Ads by Google