Reshaping the Bet v 1 fold modulates TH polarization

Slides:

Advertisements

Similar presentations

Immunomodulatory effects of aqueous birch pollen extracts and phytoprostanes on primary immune responses in vivo Jan Gutermuth, MD, Mayte Bewersdorff,

Advertisements

Birch pollen–related food allergy: Clinical aspects and the role of allergen-specific IgE and IgG4 antibodies Marija Geroldinger-Simic, MD, Thomas Zelniker,

Cor a 1–reactive T cells and IgE are predominantly cross-reactive to Bet v 1 in patients with birch pollen–associated food allergy to hazelnut Claudia.

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

Mapping of conformational IgE epitopes on Phl p 5a by using mimotopes from a phage display library Brigitte Hantusch, MSc, Sigurd Krieger, TA, Eva Untersmayr,

Luise Westernberg, PhD, Véronique Schulten, PhD, Jason A

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

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

Martin Wolf, PhD, Lorenz Aglas, PhD, Teresa E

Designing hypoallergenic derivatives for allergy treatment by means of in silico mutation and screening Theresa Thalhamer, PhD, Heidi Dobias, MSc, Tatjana.

Cor a 1–reactive T cells and IgE are predominantly cross-reactive to Bet v 1 in patients with birch pollen–associated food allergy to hazelnut Claudia.

IgE cross-reactivity between the major peanut allergen Ara h 2 and the nonhomologous allergens Ara h 1 and Ara h 3 Merima Bublin, PhD, Maria Kostadinova,

Development and characterization of a recombinant, hypoallergenic, peptide-based vaccine for grass pollen allergy Margarete Focke-Tejkl, PhD, Milena.

Blocking antibodies induced by immunization with a hypoallergenic parvalbumin mutant reduce allergic symptoms in a mouse model of fish allergy Raphaela.

Mapping of conformational IgE epitopes on Phl p 5a by using mimotopes from a phage display library Brigitte Hantusch, MSc, Sigurd Krieger, TA, Eva Untersmayr,

Elisabeth Roesler, MSc, Richard Weiss, PhD, Esther E

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

Birch pollen immunotherapy results in long-term loss of Bet v 1–specific TH2 responses, transient TR1 activation, and synthesis of IgE-blocking antibodies

Antibody conjugates bispecific for intercellular adhesion molecule 1 and allergen prevent migration of allergens through respiratory epithelial cell layers

Characterization of human memory CD4+ T-cell responses to the dog allergen Can f 4 Aino L. Rönkä, MD, Tuure T. Kinnunen, MD, PhD, Amélie Goudet, BSc,

Direct monitoring of basophil degranulation by using avidin-based probes Régis Joulia, PhD, Claire Mailhol, MD, Salvatore Valitutti, MD, Alain Didier,

Staphylococcus aureus fibronectin-binding protein specifically binds IgE from patients with atopic dermatitis and requires antigen presentation for cellular.

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

Differential expression of functional chemokine receptors on human blood and lung group 2 innate lymphoid cells Cathryn A. Weston, PhD, Batika M.J. Rana,

A fusion protein of flagellin and ovalbumin suppresses the TH2 response and prevents murine intestinal allergy Stefan Schülke, PhD, Manja Burggraf, MSc,

Naturally processed T cell–activating peptides of the major birch pollen allergen Sonja Mutschlechner, PhD, Matthias Egger, PhD, Peter Briza, PhD, Michael.

Jethe O. F. Nunes, PhD, Juliana de Souza Apostolico, MSc, David A. G

Fibronectin is a TH1-specific molecule in human subjects

Dissection of the IgE and T-cell recognition of the major group 5 grass pollen allergen Phl p 5 Margarete Focke-Tejkl, PhD, Raffaela Campana, PhD, Renate.

Allison K. Martin, BS, Douglas G. Mack, PhD, Michael T

Β2 integrins rather than β1 integrins mediate Alternaria-induced group 2 innate lymphoid cell trafficking to the lung Maya R. Karta, PhD, Peter S. Rosenthal,

Identification of bee venom Api m 1 IgE epitopes and characterization of corresponding mimotopes Abida Zahirović, MPharm, Ana Koren, PhD, Peter Kopač,

Protein unfolding strongly modulates the allergenicity and immunogenicity of Pru p 3, the major peach allergen Masako Toda, PhD, Gerald Reese, PhD, Gabriele.

Suppression of the basophil response to allergen during treatment with omalizumab is dependent on 2 competing factors Donald W. MacGlashan, MD, PhD,

Bet v 1–specific T-cell receptor/forkhead box protein 3 transgenic T cells suppress Bet v 1–specific T-cell effector function in an activation-dependent.

The T-cell response to Amb a 1 is characterized by 3 dominant epitopes and multiple MHC restriction elements Beatrice Jahn-Schmid, PhD, Nicole Wopfner,

Induction of long-lived allergen-specific plasma cells by mucosal allergen challenge Elke O. Luger, PhD, Verena Fokuhl, MSc, Michael Wegmann, PhD, Melanie.

Characterization of the allergic T-cell response to Pru p 3, the nonspecific lipid transfer protein in peach Véronique Schulten, MSc, Astrid Radakovics,

Fibronectin is a TH1-specific molecule in human subjects

Genetically engineered hybrid proteins from Parietaria judaica pollen for allergen- specific immunotherapy Roberto González-Rioja, PhD, Ignacio Ibarrola,

A hypoallergenic variant of Der p 1 as a candidate for mite allergy vaccines David Walgraffe, PhD, Christel Mattéotti, PhD, Mohamed el Bakkoury, PhD,

Kerstin Bauermeister, MSc, Barbara K

Individual IL-3 priming is crucial for consistent in vitro activation of donor basophils in patients with chronic urticaria Thomas Gentinetta, MSc, Tatjana.

Peanut T-cell epitope discovery: Ara h 1

Grass tablet sublingual immunotherapy downregulates the TH2 cytokine response followed by regulatory T-cell generation Abel Suárez-Fueyo, PhD, Tania.

Melanie Albrecht, MSc, Yvonne Kühne, MSc, Barbara K

Antigen presentation of the immunodominant T-cell epitope of the major mugwort pollen allergen, Art v 1, is associated with the expression of HLA-DRB1∗01

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

Priscilla Auyeung, PhD, Diana Mittag, PhD, Philip D

Context matters: TH2 polarization resulting from pollen composition and not from protein-intrinsic allergenicity Lorenz Aglas, PhD, Stefanie Gilles,

Novel allergic asthma model demonstrates ST2-dependent dendritic cell targeting by cypress pollen Lucia Gabriele, BS, Giovanna Schiavoni, BS, Fabrizio.

Oktay Kirak, MD, Gert Riethmüller, MD

TH1-promoting DNA immunization against allergens modulates the ratio of IgG1/IgG2a but does not affect the anaphylactic potential of IgG1 antibodies

Association between specific timothy grass antigens and changes in TH1- and TH2-cell responses following specific immunotherapy Véronique Schulten, PhD,

A hypoallergenic cat vaccine based on Fel d 1–derived peptides fused to hepatitis B PreS Katarzyna Niespodziana, MSc, Margarete Focke-Tejkl, PhD, Birgit.

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

Birch pollen–related food allergy: Clinical aspects and the role of allergen-specific IgE and IgG4 antibodies Marija Geroldinger-Simic, MD, Thomas Zelniker,

Peach allergy in China: A dominant role for mugwort pollen lipid transfer protein as a primary sensitizer Zhong-Shan Gao, PhD, Zhao-Wei Yang, BSc, Shan-Dong.

α-Purothionin, a new wheat allergen associated with severe allergy

Hypoallergenic derivatives of the major birch pollen allergen Bet v 1 obtained by rational sequence reassembly Raffaela Campana, PhD, Susanne Vrtala,

Regina Selb, PhD, Julia Eckl-Dorna, MD, PhD, Teresa E

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

Profilin (Che a 2) and polcalcin (Che a 3) are relevant allergens of Chenopodium album pollen: Isolation, amino acid sequences, and immunologic properties

Ovalbumin-specific IgE modulates ovalbumin-specific T-cell response after repetitive oral antigen administration Nemuko Omata, MD, Yusei Ohshima, MD,

Gastrointestinal digestion of Bet v 1–homologous food allergens destroys their mediator- releasing, but not T cell–activating, capacity Eva Maria Schimek,

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

Carrier-bound, nonallergenic Ole e 1 peptides for vaccination against olive pollen allergy Teresa E. Twaroch, MSc, Margit Focke, PhD, Vera Civaj, Milena.

The effects of gastric digestion on codfish allergenicity

Allergy multivaccines created by DNA shuffling of tree pollen allergens Michael Wallner, PhD, Angelika Stöcklinger, MSc, Theresa Thalhamer, MSc, Barbara.

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

Presentation transcript:

Reshaping the Bet v 1 fold modulates TH polarization Michael Wallner, PhD, Michael Hauser, PhD, Martin Himly, PhD, Nadja Zaborsky, PhD, Sonja Mutschlechner, PhD, Andrea Harrer, PhD, Claudia Asam, MSc, Ulrike Pichler, MSc, Ronald van Ree, PhD, Peter Briza, PhD, Josef Thalhamer, PhD, Barbara Bohle, PhD, Gernot Achatz, PhD, Fatima Ferreira, PhD Journal of Allergy and Clinical Immunology Volume 127, Issue 6, Pages 1571-1578.e9 (June 2011) DOI: 10.1016/j.jaci.2011.01.064 Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 A, Circular dichroism spectra of Bet v 1 and BM4 at 20°C. Spectra are baseline corrected and presented as mean residue molar ellipticity. deg, Degree; MRW, mean residue weight. B, The FTIR spectra of Bet v 1 and BM4 are presented as second derivatives. Typical regions of secondary structure elements are highlighted (α-helix, blue; intramolecular β-sheet, red; intermolecular β-sheet, green). Journal of Allergy and Clinical Immunology 2011 127, 1571-1578.e9DOI: (10.1016/j.jaci.2011.01.064) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 Aggregation behavior of Bet v 1 and BM4 were analyzed by DLS (small panel) and gel filtration (large panel). Hydrodynamic radius and molecular weight of both proteins were determined by light scattering. HP-SEC, High performance-size exclusion chromatography; MW, molecular weight; RALS, Right-angle light scattering; RH, hydrodynamic radius; RI, refractive index. Journal of Allergy and Clinical Immunology 2011 127, 1571-1578.e9DOI: (10.1016/j.jaci.2011.01.064) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 3 Proliferative responses of PBMCs from donors with birch pollen allergy stimulated with increasing amounts of Bet v 1 or BM4. Symbols represent individual patients, bars medians. P values were calculated by paired-sample t test (∗P < .05). Journal of Allergy and Clinical Immunology 2011 127, 1571-1578.e9DOI: (10.1016/j.jaci.2011.01.064) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 4 A, BALB/c mice were immunized 4 times with either Bet v 1 (filled circles) or BM4 (open circles; n = 8 per group). Bet v 1–specific antibody levels were determined by ELISA. B, ELISPOT analysis of splenocytes from immunized animals expressed as mean cytokine-secreting cells per 105 spleen cells ± SEM. P values were calculated with t tests (∗∗P < .01; ∗∗∗P < .001). Journal of Allergy and Clinical Immunology 2011 127, 1571-1578.e9DOI: (10.1016/j.jaci.2011.01.064) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 5 BMDCs were pulsed with FITC or pHrodo-labeled Bet v 1 or BM4, stained for CD11c and CD86, and gated for CD11c expression. Representative fluorescence-activated cell-sorting profiles of 6 experiments per time point and antigen are shown. Numbers in the panels represent percentage values of analyzed cells (left panel). A time course analysis is presented as a line chart ± SEM (right panel). P values were calculated with t tests (∗P < .05; ∗∗P < .01). Journal of Allergy and Clinical Immunology 2011 127, 1571-1578.e9DOI: (10.1016/j.jaci.2011.01.064) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 6 A, SDS-PAGE analysis of time-dependent endo-/lysosomal degradation of Bet v 1 and BM4. B, Percent degradation was determined densitometrically from SDS-PAGE bands and is presented as a line chart. C, Proteolytic fragments of Bet v 1 (black lines) or BM4 (red lines) were analyzed by mass spectrometry. Dominant T cell epitopes of Bet v 1 are indicated by vertical blue bars. Journal of Allergy and Clinical Immunology 2011 127, 1571-1578.e9DOI: (10.1016/j.jaci.2011.01.064) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Primary sequence of BM4 compared to Bet v 1, isoform 0101, and Mal d 1, isoform 0108. The exchange sequence of BM4 is highlighted by a gray box. Journal of Allergy and Clinical Immunology 2011 127, 1571-1578.e9DOI: (10.1016/j.jaci.2011.01.064) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

ANS binding to Bet v 1 and BM4 was analyzed with fluorescence spectroscopy. Data are presented in mean fluorescence units (RFU). Journal of Allergy and Clinical Immunology 2011 127, 1571-1578.e9DOI: (10.1016/j.jaci.2011.01.064) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

IgE binding of BM4 was compared with Bet v 1 by ELISA detecting antigen-specific human serum IgE (n = 13; A), RBL assays using patients’ sera (n = 8; B), or basophil activation tests (n = 8; B). C, Four characteristic RBL titration curves are shown. Bet v 1 is represented as a black line and BM4 as a red line in the graphs. OVA, Ovalbumin. Journal of Allergy and Clinical Immunology 2011 127, 1571-1578.e9DOI: (10.1016/j.jaci.2011.01.064) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

BALB/c mice were immunized 4 times with Bet v 1∗ (filled circle), A1-6 (filled triangle), or BM4 (open circle; n = 5 per group). Bet v 1∗–specific antibody levels were determined by ELISA. P values were calculated with t tests (∗P < .05). Journal of Allergy and Clinical Immunology 2011 127, 1571-1578.e9DOI: (10.1016/j.jaci.2011.01.064) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

BALB/c mice were immunized 4 times with Bet v 1∗ (black bars), A1-6 (gray bars), or BM4 (white bars; n = 5 per group). ELISPOT analysis of splenocytes from immunized animals expressed as mean cytokine-secreting cells per 2 × 105 spleen cells ± SEM. P values were calculated with t tests (∗P < .05; ∗∗P < .01; ∗∗∗P < .001). Journal of Allergy and Clinical Immunology 2011 127, 1571-1578.e9DOI: (10.1016/j.jaci.2011.01.064) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Bet v 1–coated ELISA plates were preincubated with IgE-depleted pooled mouse sera of animals immunized with Bet v 1 (n = 5) or BM4 (n = 5), and human serum IgE binding of 5 patients with birch pollen allergy was determined. The threshold was calculated by preincubation of Bet v 1 with pooled murine preimmune sera (n = 5). Journal of Allergy and Clinical Immunology 2011 127, 1571-1578.e9DOI: (10.1016/j.jaci.2011.01.064) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions