Lentiviral-mediated gene therapy leads to improvement of B-cell functionality in a murine model of Wiskott-Aldrich syndrome  Marita Bosticardo, PhD, Elena.

Slides:



Advertisements
Similar presentations
Differentiation stage determines pathologic and protective allergen-specific CD4+ T-cell outcomes during specific immunotherapy  Erik Wambre, PhD, Jonathan.
Advertisements

Serum amyloid P attenuates M2 macrophage activation and protects against fungal spore–induced allergic airway disease  Ana Paula Moreira, PhD, Karen A.
Immacolata Brigida, PhD, Aisha V
The loss of IgM memory B cells correlates with clinical disease in common variable immunodeficiency  Rita Carsetti, MD, Maria Manuela Rosado, PhD, Simona.
Birch pollen–related food allergy: Clinical aspects and the role of allergen-specific IgE and IgG4 antibodies  Marija Geroldinger-Simic, MD, Thomas Zelniker,
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,
The importance of being “pure” neutrophils
Effects of hyperprolactinemia treatment with the dopamine agonist quinagolide on endometriotic lesions in patients with endometriosis-associated hyperprolactinemia 
Cell-specific activation profile of extracellular signal-regulated kinase 1/2, Jun N-terminal kinase, and p38 mitogen-activated protein kinases in asthmatic.
Delivery of antigen to nasal-associated lymphoid tissue microfold cells through secretory IgA targeting local dendritic cells confers protective immunity 
Regulatory B cells prevent and reverse allergic airway inflammation via FoxP3-positive T regulatory cells in a murine model  Sylvie Amu, PhD, Sean P.
B-cell reconstitution after lentiviral vector–mediated gene therapy in patients with Wiskott-Aldrich syndrome  Maria Carmina Castiello, PhD, Samantha.
Jay A. Lieberman, MD, Faith R. Huang, MD, Hugh A
Intravenous immunoglobulin treatment abrogates transplacental autoantibody transfer in a murine pemphigus model  Sachiko Ono, MD, PhD, Gyohei Egawa, MD,
Interferon response factor 3 is essential for house dust mite–induced airway allergy  Thomas Marichal, DVM, Denis Bedoret, DVM, PhD, Claire Mesnil, DVM,
Ting-ting Zhang, MSc, Klaus Okkenhaug, PhD, Baher F
Humanized mouse model of mast cell–mediated passive cutaneous anaphylaxis and passive systemic anaphylaxis  Paul J. Bryce, PhD, Rustom Falahati, PhD,
Frank Kirstein, PhD, Natalie E
Magdalena A. Berkowska, PhD, Jorn J
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,
Assessing basophil activation by using flow cytometry and mass cytometry in blood stored 24 hours before analysis  Kaori Mukai, PhD, Nicolas Gaudenzio,
Targeting lentiviral vector expression to hepatocytes limits transgene-specific immune response and establishes long-term expression of human antihemophilic.
In vivo T-cell dynamics during immune reconstitution after hematopoietic stem cell gene therapy in adenosine deaminase severe combined immune deficiency 
Neonatal rhinovirus induces mucous metaplasia and airways hyperresponsiveness through IL-25 and type 2 innate lymphoid cells  Jun Young Hong, MS, J. Kelley.
Cell-specific activation profile of extracellular signal-regulated kinase 1/2, Jun N-terminal kinase, and p38 mitogen-activated protein kinases in asthmatic.
Leukocyte nicotinamide adenine dinucleotide phosphate-reduced oxidase is required for isocyanate-induced lung inflammation  Si-Yen Liu, PhD, Wei-Zhi Wang,
Birch pollen immunotherapy results in long-term loss of Bet v 1–specific TH2 responses, transient TR1 activation, and synthesis of IgE-blocking antibodies 
Recombination-activating gene 1 (Rag1)–deficient mice with severe combined immunodeficiency treated with lentiviral gene therapy demonstrate autoimmune.
Signaling through FcRγ-associated receptors on dendritic cells drives IL-33–dependent TH2-type responses  Melissa Y. Tjota, BA, Cara L. Hrusch, PhD, Kelly.
The BLNK adaptor protein has a nonredundant role in human B-cell differentiation  Chantal Lagresle-Peyrou, PhD, Michèle Millili, Sonia Luce, Annie Boned,
Dorien Van Saen, Ph. D. , Ellen Goossens, Ph. D. , Joeri L. Aerts, Ph
Ζ Chain–associated protein of 70 kDa (ZAP70) deficiency in human subjects is associated with abnormalities of thymic stromal cells: Implications for T-cell.
Frank Kirstein, PhD, Natalie E
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.
Colony-Stimulating Factor-1-Dependent Macrophages Are Responsible for IVIG Protection in Antibody-Induced Autoimmune Disease  Pierre Bruhns, Astrid Samuelsson,
B-1a and B-1b Cells Exhibit Distinct Developmental Requirements and Have Unique Functional Roles in Innate and Adaptive Immunity to S. pneumoniae  Karen.
Defective B-cell memory in patients with Down syndrome
Endogenous polyclonal anti–IL-1 antibody responses potentiate IL-1 activity during pathogenic inflammation  Gunther Spohn, PhD, Natalia Arenas-Ramirez,
Revertant T lymphocytes in a patient with Wiskott-Aldrich syndrome: Analysis of function and distribution in lymphoid organs  Sara Trifari, PhD, Samantha.
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,
A peptide derived from the Wiskott-Aldrich syndrome (WAS) protein-interacting protein (WIP) restores WAS protein level and actin cytoskeleton reorganization.
Peanut-induced intestinal allergy is mediated through a mast cell–IgE–FcεRI–IL-13 pathway  Meiqin Wang, MD, PhD, Katsuyuki Takeda, MD, PhD, Yoshiki Shiraishi,
Volume 29, Issue 2, Pages (August 2008)
Role of B cells in TH cell responses in a mouse model of asthma
Volume 10, Issue 6, Pages (December 2004)
Mammalian target of rapamycin inhibition counterbalances the inflammatory status of immune cells in patients with chronic granulomatous disease  Aurélie.
Chen Yao, BS, Sandra M. Zurawski, PhD, Elizabeth S
Toward experimental assessment of receptor occupancy: TGN1412 revisited  Zoe Waibler, PhD, Linda Y. Sender, Christel Kamp, PhD, Jan Müller-Berghaus, MD,
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,
Oral administration of a synthetic agonist of Toll-like receptor 9 potently modulates peanut-induced allergy in mice  Fu-Gang Zhu, PhD, Ekambar R. Kandimalla,
Differentiation stage determines pathologic and protective allergen-specific CD4+ T-cell outcomes during specific immunotherapy  Erik Wambre, PhD, Jonathan.
IL-25 as a novel therapeutic target in nasal polyps of patients with chronic rhinosinusitis  Hyun-Woo Shin, MD, PhD, Dong-Kyu Kim, MD, Min-Hyun Park, MD,
Xin-Zi Tang, PhD, James B. Jung, BS, Christopher D.C. Allen, PhD 
Langerhans cells are critical in epicutaneous sensitization with protein antigen via thymic stromal lymphopoietin receptor signaling  Saeko Nakajima,
Reshaping the Bet v 1 fold modulates TH polarization
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
Allergen-specific immunotherapy modulates the balance of circulating Tfh and Tfr cells  Véronique Schulten, PhD, Victoria Tripple, BSc, Grégory Seumois,
Birch pollen–related food allergy: Clinical aspects and the role of allergen-specific IgE and IgG4 antibodies  Marija Geroldinger-Simic, MD, Thomas Zelniker,
Volume 21, Issue 1, Pages (January 2013)
Serum amyloid P attenuates M2 macrophage activation and protects against fungal spore–induced allergic airway disease  Ana Paula Moreira, PhD, Karen A.
Journal of Allergy and Clinical Immunology
Mice deficient in the St3gal3 gene product α2,3 sialyltransferase (ST3Gal-III) exhibit enhanced allergic eosinophilic airway inflammation  Takumi Kiwamoto,
CD11b-mediated migratory property of peripheral blood B cells
Volume 17, Issue 6, Pages (June 2009)
Volume 20, Issue 10, Pages (October 2012)
Mice deficient in the St3gal3 gene product α2,3 sialyltransferase (ST3Gal-III) exhibit enhanced allergic eosinophilic airway inflammation  Takumi Kiwamoto,
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:

Lentiviral-mediated gene therapy leads to improvement of B-cell functionality in a murine model of Wiskott-Aldrich syndrome  Marita Bosticardo, PhD, Elena Draghici, MS, Francesca Schena, PhD, Aisha Vanessa Sauer, PhD, Elena Fontana, PhD, Maria Carmina Castiello, MS, Marco Catucci, PhD, Michela Locci, PhD, Luigi Naldini, MD, PhD, Alessandro Aiuti, MD, PhD, Maria Grazia Roncarolo, MD, Pietro Luigi Poliani, MD, PhD, Elisabetta Traggiai, PhD, Anna Villa, MD  Journal of Allergy and Clinical Immunology  Volume 127, Issue 6, Pages 1376-1384.e5 (June 2011) DOI: 10.1016/j.jaci.2011.03.030 Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 Representative FACS histograms showing WASp staining in each B-cell subset analyzed in BM (A, from left to right: pre–B-cell, immature B-cell, and mature/recirculating B-cell subsets), peritoneal cavity (B, from left to right: B1a-cell, B1b-cell, and B2-cell subsets), and spleen (C, from left to right: transitional B-cell, follicular B-cell, and MZ B-cell subsets) of BMT wt (gray clear histograms), BMT was−/− (gray filled histograms) and GT mice (black clear histograms). In each panel, the positioning of the markers used to calculate frequency and MFI of WASp+ cells is also shown. Max, Maximum. Journal of Allergy and Clinical Immunology 2011 127, 1376-1384.e5DOI: (10.1016/j.jaci.2011.03.030) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 Evidence of WASp expression in B-cell subsets from tissues of GT-treated mice. Frequency of WASp+ cells in Was−/− mice transplanted with Lin− BM wt cells (BMT wt), Was−/− cells untransduced (BMT was−/−) or LV-transduced (GT). A, BM. Percentage of WASp+ cells among pre-B, immature B, and mature/recirculating (recirc) B cells. B, Peritoneal cavity. Percentage of WASp+ cells among B1a cells, B1b cells, and mature follicular (B2) cells. C, Spleen. Percentage of WASp+ cells among transitional and mature B cells (MZ and follicular). Results shown were obtained by analyzing 15 to 34 mice per group. Median values are shown per each group. The Mann-Whitney test was used to perform statistical analysis of data. ∗P < .05; ∗∗∗P < .001. Journal of Allergy and Clinical Immunology 2011 127, 1376-1384.e5DOI: (10.1016/j.jaci.2011.03.030) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 3 MFI of WASp expression in B-cell subsets from tissues of GT-treated mice. MFI of WASp was evaluated in Was−/− mice transplanted with Lin− BM wt cells (BMT wt), Was−/− cells untransduced (BMT was−/−) or LV-transduced (GT). In the graphs, MFI is expressed as fold increase in respect to values found in the BMT was−/− group. In the GT group, MFI was calculated only on WASp+ cells. A, BM (pre B, immature B, and mature/recirculating [recirc] B-cell subsets). B, Peritoneal cavity (B1a cells, B1b cells, and B2 cells). C, Spleen (transitional, follicular [FO], and MZ B cells). Results shown were obtained by analyzing 15 to 32 mice per group. Median values are shown for each group. The Mann-Whitney test was used to perform statistical analysis of data. ∗∗P < .005; ∗∗∗P < .001. Journal of Allergy and Clinical Immunology 2011 127, 1376-1384.e5DOI: (10.1016/j.jaci.2011.03.030) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 4 Presence of WASp-expressing cells in splenic tissue sections from GT-treated Was−/− mice. Spleen sections from BMT wt (A), BMT was−/− (B), and GT mice (C) were stained with hematoxylin and eosin (H&E; left panels), anti-WASp (middle panels), or anti-B220 antibody (right panels). All images are ×10 original magnification. Journal of Allergy and Clinical Immunology 2011 127, 1376-1384.e5DOI: (10.1016/j.jaci.2011.03.030) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 5 Improvement of splenic B-cell follicle and MZ architecture in GT-treated Was−/− mice. Spleen sections from BMT wt (A), BMT was−/− (B), and GT mice (C) were stained for antimacrophage scavenger receptor MARCO to identify MZMs surrounding spleen follicles, highlighted by anti-B220 immunostaining (left panels; ×10 original magnification). MZ B cells (IgMhiIgDlow/-, indicated by arrows) have been highlighted by double immunostainings for anti-MoMa (blue signal) combined with either IgD or IgM (brown signals; right panels; ×20 original magnification). Journal of Allergy and Clinical Immunology 2011 127, 1376-1384.e5DOI: (10.1016/j.jaci.2011.03.030) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 6 GT increases response to polysaccharide antigens in Was−/− mice. In vivo challenge of treated mice was performed by intraperitoneal injection of 0.5 μg/mouse Pneumovax23. IgM Pneumovax23-specific antibodies were evaluated by ELISA assay on serum collected 7 days after immunization. The results shown in the graph were obtained from BMT wt (n = 23; black circles), BMT was−/− (n = 14; white circles), and GT mice (n = 20; gray squares). Statistical analysis was performed by using a 2-way ANOVA test. ∗P < .05; ∗∗∗P < .001. Journal of Allergy and Clinical Immunology 2011 127, 1376-1384.e5DOI: (10.1016/j.jaci.2011.03.030) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 7 GT results in decreased autoantibody titer in Was−/− mice. A, The presence of anti-dsDNA autoantibodies was evaluated by ELISA assay. The graph shows the frequency of positivity (titer ≥ 1:60) for anti-dsDNA autoantibodies in the 3 groups of treated mice (BMT wt, black bar; BMT was−/−, white bar; GT, gray bar). The number of mice analyzed per group is shown in brackets below each bar. Statistical analysis was performed using a Fisher exact t test. ∗P < .05; ∗∗∗P < .001. B, The graph shows the OD values at serial dilutions (from 1:20 to 1:640) restricted to serum samples that were positive for the presence of anti-dsDNA autoantibodies (BMT was−/−, n = 10, white circles; GT, n = 7, gray circles). Statistical analysis was performed by using a 2-way ANOVA test. ∗∗P < .005. C, Tissue-specific autoantibodies were detected by indirect immunohistochemistry on cryostat sections (7 μm) from adult Rag2−/−γc−/− mice. Each serum was analyzed for the reactivity to 3 tissues (salivary gland, thyroid, gastrointestinal tract). The graph shows the frequency of serum samples reactive to 0, 1, 2, or 3 tissues for the 3 groups of mice. The number of samples analyzed is indicated in brackets below each bar. ∗∗∗P < .001. D, The figure shows autoantibodies against salivary gland in mice representative of each group of treatment (BMT wt, BMT was−/−, and GT). Magnification ×40. Journal of Allergy and Clinical Immunology 2011 127, 1376-1384.e5DOI: (10.1016/j.jaci.2011.03.030) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E1 Presence of gene corrected cells in BM and spleen (SP) in Was−/− mice reconstituted with LV-transduced Lin− BM cells. Lentiviral VCN/cell was evaluated on total BM and SP cells. Results shown were obtained analyzing 38 GT mice. Median value is shown per each group. Wilcoxon signed-rank test was used to perform statistical analysis of data. ∗∗∗P < .001. Journal of Allergy and Clinical Immunology 2011 127, 1376-1384.e5DOI: (10.1016/j.jaci.2011.03.030) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E2 Analysis of WASp-expressing cell frequency in peripheral blood B220+ cells from GT-treated mice 3 and 9 months after transplant. ∗∗P < .005 (paired t test). Journal of Allergy and Clinical Immunology 2011 127, 1376-1384.e5DOI: (10.1016/j.jaci.2011.03.030) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E3 Presence of WASp-expressing B cells in splenic tissue sections from GT-treated Was−/− mice. Spleen sections from BMT wt (A), BMT was−/− (B), and GT mice (C) have been stained with anti-B220 antibody (left panels; green) and anti-WASp (middle panels; red). Right panels show merging of B220 and WASp stainings (yellow). All images are from ×20 original magnification. Journal of Allergy and Clinical Immunology 2011 127, 1376-1384.e5DOI: (10.1016/j.jaci.2011.03.030) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions