Extracellular vesicle–derived protein from Bifidobacterium longum alleviates food allergy through mast cell suppression Jung-Hwan Kim, PhD, Eun-Ji Jeun,

Slides:

Advertisements

Similar presentations

Alternation of circadian clock modulates forkhead box protein-3 gene transcription in CD4+ T cells in the intestine Gui Yang, MD, Hong Zhang, PhD, Yi.

Advertisements

Therapeutic reversal of food allergen sensitivity by mature retinoic acid–differentiated dendritic cell induction of LAG3+CD49b−Foxp3− regulatory T cells

Thymic stromal lymphopoietin signaling in CD4+ T cells is required for TH2 memory Qun Wang, PhD, Jianguang Du, PhD, Jingjing Zhu, MSc, Xiaowei Yang, MSc,

Allergen endotoxins induce T-cell–dependent and non–IgE-mediated nasal hypersensitivity in mice Naruhito Iwasaki, MD, Kazufumi Matsushita, PhD, Ayumi.

IgE-mediated systemic anaphylaxis and impaired tolerance to food antigens in mice with enhanced IL-4 receptor signaling Clinton B. Mathias, PhD, Suejy.

The activating protein 1 transcription factor basic leucine zipper transcription factor, ATF- like (BATF), regulates lymphocyte- and mast cell–driven immune.

Regulatory B cells prevent and reverse allergic airway inflammation via FoxP3-positive T regulatory cells in a murine model Sylvie Amu, PhD, Sean P.

The activating protein 1 transcription factor basic leucine zipper transcription factor, ATF- like (BATF), regulates lymphocyte- and mast cell–driven immune.

Alternation of circadian clock modulates forkhead box protein-3 gene transcription in CD4+ T cells in the intestine Gui Yang, MD, Hong Zhang, PhD, Yi.

IL-25 and CD4+ TH2 cells enhance type 2 innate lymphoid cell–derived IL-13 production, which promotes IgE-mediated experimental food allergy Jee-Boong.

Allergy prevention starts before conception: Maternofetal transfer of tolerance protects against the development of asthma Tobias Polte, PhD, Christian.

Selective control of SIRP-α–positive airway dendritic cell trafficking through CD47 is critical for the development of TH2-mediated allergic inflammation

Oral immunotherapy induces IgG antibodies that act through FcγRIIb to suppress IgE- mediated hypersensitivity Oliver T. Burton, PhD, Stephanie L. Logsdon,

Frank Kirstein, PhD, Natalie E

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,

Pentraxin 3 deletion aggravates allergic inflammation through a TH17-dominant phenotype and enhanced CD4 T-cell survival Jyoti Balhara, MSc, Lianyu Shan,

Epicutaneous sensitization results in IgE-dependent intestinal mast cell expansion and food-induced anaphylaxis Lisa M. Bartnikas, MD, Michael F. Gurish,

Compartmentalized chemokine-dependent regulatory T-cell inhibition of allergic pulmonary inflammation Roshi Afshar, PhD, James P. Strassner, BS, Edward.

Matthew C. Tunis, BSc, Wojciech Dawicki, PhD, Kaitlyn R

IL-33 dysregulates regulatory T cells and impairs established immunologic tolerance in the lungs Chien-Chang Chen, PhD, Takao Kobayashi, PhD, Koji Iijima,

Restoration of T-box–containing protein expressed in T cells protects against allergen- induced asthma Jung Won Park, MD, Hyun Jung Min, MS, Jung Ho Sohn,

Chronic rhinosinusitis with polyps and without polyps is associated with increased expression of suppressors of cytokine signaling 1 and 3 Se Jin Park,

Oral Administration of Poly-γ-Glutamate Ameliorates Atopic Dermatitis in Nc/Nga Mice by Suppressing Th2-Biased Immune Response and Production of IL-17A

Color-coded real-time cellular imaging of lung T-lymphocyte accumulation and focus formation in a mouse asthma model Akihiro Hasegawa, PhD, Katsuhiro.

Signaling through FcRγ-associated receptors on dendritic cells drives IL-33–dependent TH2-type responses Melissa Y. Tjota, BA, Cara L. Hrusch, PhD, Kelly.

Activin A and TGF-β promote TH9 cell–mediated pulmonary allergic pathology Carla P. Jones, PhD, Lisa G. Gregory, PhD, Benjamin Causton, BSc, Gaynor A.

Basophil-derived IL-4 promotes epicutaneous antigen sensitization concomitant with the development of food allergy Maryam Hussain, MSc, Loïc Borcard,

Curcumin, a constituent of curry, suppresses IgE-mediated allergic response and mast cell activation at the level of Syk Jun Ho Lee, PhD, Jie Wan Kim,

Pharmacologic inhibition of Notch signaling suppresses food antigen–induced mucosal mast cell hyperplasia Asuka Honjo, MD, Nobuhiro Nakano, PhD, Susumu.

Early-life chlamydial lung infection enhances allergic airways disease through age- dependent differences in immunopathology Jay C. Horvat, PhD, Malcolm.

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

Differential roles for the IL-9/IL-9 receptor α-chain pathway in systemic and oral antigen–induced anaphylaxis Heather Osterfeld, BSc, Richard Ahrens,

Oliver T. Burton, PhD, Jaciel M. Tamayo, PhD, Amanda J

Experimental gastrointestinal allergy enhances pulmonary responses to specific and unrelated allergens Eric B. Brandt, PhD, Troy A. Scribner, MD, Hiroko.

Frank Kirstein, PhD, Natalie E

Ton C. Doan, BA, Brian M. Jeong, MSc, Mackenzie E. Coden, BA, Lucas F

T-cell immunoglobulin and mucin domain 1 deficiency eliminates airway hyperreactivity triggered by the recognition of airway cell death Hye Young Kim,

Exposure to food allergens through inflamed skin promotes intestinal food allergy through the thymic stromal lymphopoietin–basophil axis Mario Noti,

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

Prostaglandin E2 suppresses allergic sensitization and lung inflammation by targeting the E prostanoid 2 receptor on T cells Zbigniew Zasłona, PhD, Katsuhide.

Food Allergy Herbal Formula-2 silences peanut-induced anaphylaxis for a prolonged posttreatment period via IFN-γ–producing CD8+ T cells Kamal D. Srivastava,

Exaggerated IL-17 response to epicutaneous sensitization mediates airway inflammation in the absence of IL-4 and IL-13 Rui He, MD, PhD, Hye Young Kim,

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

IL-33 promotes food anaphylaxis in epicutaneously sensitized mice by targeting mast cells Claire Galand, PhD, Juan Manuel Leyva-Castillo, PhD, Juhan.

3-Methyl-4-nitrophenol triggers nasal allergy by modulating dendritic cell properties Xiao-Yu Liu, PhD, Yong-Jin Wu, MD, PhD, Li-Juan Song, MD, Xian-Hai.

CD4-mediated regulatory T-cell activation inhibits the development of disease in a humanized mouse model of allergic airway disease Helen Martin, MS,

Docosahexaenoic Acid Alleviates Atopic Dermatitis by Generating Tregs and IL-10/TGF- β-Modified Macrophages via a TGF-β-Dependent Mechanism Sang-Chul.

Mechanisms underlying differential food allergy response to heated egg

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,

Basophil-derived IL-4 promotes epicutaneous antigen sensitization concomitant with the development of food allergy Maryam Hussain, MSc, Loïc Borcard,

Thymic stromal lymphopoietin–activated invariant natural killer T cells trigger an innate allergic immune response in atopic dermatitis Wen Hao Wu, PhD,

Prostaglandin E2 suppresses allergic sensitization and lung inflammation by targeting the E prostanoid 2 receptor on T cells Zbigniew Zasłona, PhD, Katsuhide.

Role of B cells in TH cell responses in a mouse model of asthma

T-bet inhibits innate lymphoid cell–mediated eosinophilic airway inflammation by suppressing IL-9 production Ayako Matsuki, MD, Hiroaki Takatori, MD,

Sarita Sehra, PhD, Weiguo Yao, PhD, Evelyn T. Nguyen, MS, Nicole L

T-bet inhibits innate lymphoid cell–mediated eosinophilic airway inflammation by suppressing IL-9 production Ayako Matsuki, MD, Hiroaki Takatori, MD,

Ana Belén Blázquez, Lloyd Mayer, M. Cecilia Berin Gastroenterology

Kirthana Ganeshan, BS, Colleen V

Recombinant basic fibroblast growth factor inhibits the airway hyperresponsiveness, mucus production, and lung inflammation induced by an allergen challenge

Oral Administration of Poly-γ-Glutamate Ameliorates Atopic Dermatitis in Nc/Nga Mice by Suppressing Th2-Biased Immune Response and Production of IL-17A

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

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,

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

Duy Pham, PhD, Sarita Sehra, PhD, Xin Sun, PhD, Mark H. Kaplan, PhD

Eric B. Brandt, PhD, Ariel Munitz, PhD, Tatyana Orekov, MS, Melissa K

The steroidogenic enzyme Cyp11a1 is essential for development of peanut-induced intestinal anaphylaxis Meiqin Wang, MD, PhD, Julita Ramirez, DVM, PhD,

Dicer is indispensable for the development of murine mast cells

Epicutaneous immunization with ovalbumin and CpG induces TH1/TH17 cytokines, which regulate IgE and IgG2a production Monika Majewska-Szczepanik, PhD,

Soybean isoflavones regulate dendritic cell function and suppress allergic sensitization to peanut Madhan Masilamani, PhD, John Wei, BA, Shiven Bhatt,

Recombinant basic fibroblast growth factor inhibits the airway hyperresponsiveness, mucus production, and lung inflammation induced by an allergen challenge

Presentation transcript:

Extracellular vesicle–derived protein from Bifidobacterium longum alleviates food allergy through mast cell suppression Jung-Hwan Kim, PhD, Eun-Ji Jeun, MSc, Chun-Pyo Hong, PhD, Seong-Hoon Kim, MSc, Min Seong Jang, DVM, PhD, Eun-Jung Lee, MSc, Sook Jin Moon, BSc, Chang Ho Yun, PhD, Sin-Hyeog Im, PhD, Seok-Geun Jeong, PhD, Beom-Young Park, PhD, Kyong-Tai Kim, PhD, Ju-Young Seoh, MD, PhD, Yoon-Keun Kim, MD, PhD, Sung-Jong Oh, PhD, Jun-Sang Ham, PhD, Bo-Gie Yang, PhD, Myoung Ho Jang, PhD Journal of Allergy and Clinical Immunology Volume 137, Issue 2, Pages 507-516.e8 (February 2016) DOI: 10.1016/j.jaci.2015.08.016 Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 Effects of B longum KACC 91563 and E faecalis KACC 91532 on food allergy–induced diarrhea. Induction of food allergy was evaluated based on diarrhea occurrence in mice. A, Experimental schedule for food allergy induction in mice. i.g., Intragastric; i.p., intraperitoneal. B, For food allergy induction, 50 mg of OVA was orally administered 5 times. Combined data from 3 independent experiments (OVA vs OVA + B longum: **P < .01, n = approximately 15-17 mice per group). Journal of Allergy and Clinical Immunology 2016 137, 507-516.e8DOI: (10.1016/j.jaci.2015.08.016) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 B longum KACC 91563 reduced the number of mast cells in the LP of the small intestine in food allergy–induced mice. A, Paraffin sections of the jejunum of the small intestine were stained for mast cells. A magnification of the LP clearly shows mast cells. The number of mast cells was counted in 4 different fields (n = 5 mice per group; original magnification ×400). HPF, High-power field. B, MCPT-1 levels in serum (n = approximately 6-7 per group). C, Levels of OVA-specific IgE in serum. Values represent means ± SEMs. One-way ANOVA with the Newman-Keuls posttest is applied. Statistical significance: *P < .05 and ***P < .001. Data are representative of 2 to 3 independent experiments. B. l, B longum KACC 91563; E. f, E faecalis KACC 91532. Journal of Allergy and Clinical Immunology 2016 137, 507-516.e8DOI: (10.1016/j.jaci.2015.08.016) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 3 Apoptosis of BMMCs induced by B longum KACC 91563 and its EVs. A, TEM image of EVs derived from B longum KACC 91563 and E faecalis KACC 91532. B, Size of EVs. Error bars indicate means ± SDs from 5 measurements. C, Apoptosis assay. Annexin V+ apoptotic cells were measured by means of fluorescence-activated cell sorting (n = 3). Values represent means ± SEMs. Statistical analysis was performed by using 1-way ANOVA with the Newman-Keuls posttest. **P < .01 and ***P < .001. Data are representative of 2 to 3 independent experiments. D, BMMCs were incubated with whole bacteria or EVs of B longum KACC 91563 or E faecalis KACC 91532 for 2 hours. Apoptotic cells were stained with the DeadEnd Fluorometric TUNEL system. Representative data are shown (magnification ×400). Nuclei of cells are blue, and TUNEL is green. The number of apoptotic cells was counted in 7 different fields (magnification ×100, n = 3 per group). Values represent means ± SEMs. Statistical analysis was performed by using 1-way ANOVA with Newman-Keuls posttest. *P < .05 and **P < .01. B. l, B longum KACC 91563; E. f, E faecalis KACC 91532. Journal of Allergy and Clinical Immunology 2016 137, 507-516.e8DOI: (10.1016/j.jaci.2015.08.016) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 4 EVs of B longum KACC 91563 and E faecalis KACC 91532 were internalized by BMMCs. A, Individual cell populations were incubated with specific bacteria-derived EVs at the indicated temperature. The percentages of cells that internalized EVs are represented with a bar graph. ns, Not significant. B, Representative images are shown. Nuclei of cells are blue, and DiI-labeled EVs are red. Values represent means ± SEMs. The Student t test was applied. Statistical significance: **P < .01. The number of cells was counted in 25 different fields was shown (n = approximately 3-4 per group; original magnification ×400). Data are representative of 2 independent experiments. B. l, B longum KACC 91563; E. f, E faecalis KACC 91532. Journal of Allergy and Clinical Immunology 2016 137, 507-516.e8DOI: (10.1016/j.jaci.2015.08.016) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 5 Suppression of food allergy responses through ESBP, a major protein of B longum KACC 91563–derived EVs. A, Proteins of B longum KACC 91563 and its EVs were analyzed by using SDS-PAGE. ESBP and ASBP were identified through proteomic analysis. B, Experimental schedule for food allergy induction and administrations of ESBP or ASBP in mice. C, Abilities of GST-tagged recombinant ESBP or ASBP (50 μg) to suppress allergic diarrhea (OVA + GST vs OVA + ESBP: ***P < .001, n = approximately 6-22 mice per group). Recombinant GST protein–treated mice were used as a positive control. Data are combined from 3 independent experiments. D, Paraffin-embedded sections of jejunum were stained for mast cells (original magnification ×400). E, The number of mast cells was counted in 4 different fields and represented by a bar graph (n = 4 mice per group). Statistical analysis was performed by using 1-way ANOVA with the Newman-Keuls posttest. *P < .05. F, Levels of MCPT-1 in serum (n = approximately 3-6 per group). Data are representative of 2 independent experiments. i.g., Intragastric; i.p., intraperitoneal. Journal of Allergy and Clinical Immunology 2016 137, 507-516.e8DOI: (10.1016/j.jaci.2015.08.016) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 6 Proposed mechanism of attenuation of food allergy by B longum KACC 91563. B longum KACC 91563 in the intestinal lumen secretes small-sized EVs that contain bacterial components. The EVs penetrate through intestinal epithelial cells and specifically induce apoptosis of mast cells in the LP. The reduction of mast cells results in suppression of food allergy. Journal of Allergy and Clinical Immunology 2016 137, 507-516.e8DOI: (10.1016/j.jaci.2015.08.016) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E1 Characterization of B longum KACC 91563 and E faecalis KACC 91532. A, Morphologic features of B longum KACC91563 and E faecalis KACC 91532 were analyzed by using TEM. B, B longum KACC91563 and E faecalis KACC 91532 were administered for 2 weeks, and FISH analysis was performed on fecal samples. Journal of Allergy and Clinical Immunology 2016 137, 507-516.e8DOI: (10.1016/j.jaci.2015.08.016) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E2 Effects of B longum KACC 91563 and E faecalis KACC 91532 on the low-dose food allergy model. For food allergy induction, 10 mg of OVA was orally administered 7 times (OVA vs OVA + B longum: *P < .05, n = approximately 6-8 mice per group). Journal of Allergy and Clinical Immunology 2016 137, 507-516.e8DOI: (10.1016/j.jaci.2015.08.016) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E3 Alleviation of food allergy by B. longum KACC 91563 was not associated with T cell–mediated immune responses. A, After the fifth intragastric challenge of OVA, MLN cells were cultured with OVA for 72 hours, and concentrations of IL-4, IL-5, IL-9, IL-10, IL-13, IL-17, and IFN-γ in the culture supernatants were measured. B, Cells were isolated from spleens and MLNs and stained for CD4 and Foxp3. Proportions of Foxp3+ Treg cells were assessed by using fluorescence-activated cell sorting. Values represent means ± SEMs (n = 4 per group). Data are representative of 2 independent experiments. B. l, B longum KACC 91563; E. f, E faecalis KACC 91532. Journal of Allergy and Clinical Immunology 2016 137, 507-516.e8DOI: (10.1016/j.jaci.2015.08.016) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E4 Effect of B longum KACC 91563 on mast cells in the intestine under homeostatic conditions. B longum KACC 91563 was administered into mice without induction of food allergy. A, Intestinal mast cell population was assessed by using fluorescence-activated cell sorting (n = 5 mice per group). B, Paraffin-embedded sections of the jejunum were stained for mast cells, and the number of mast cells was counted in 3 different fields (n = approximately 4-5 mice per group; original magnification ×400). ND, Normal diet. Values represent means ± SEMs. Journal of Allergy and Clinical Immunology 2016 137, 507-516.e8DOI: (10.1016/j.jaci.2015.08.016) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E5 Effect of acetate, a metabolite of B longum, on food allergy. A and C, Experimental schedule for food allergy induction and acetate administration in mice. B, Acetate (150 mmol/L) was administered in drink to mice every day from day 19 of the food allergy model and occurrence of allergic diarrhea was analyzed to examine the effect of acetate on food allergy symptoms (n = 5 mice per group). D, Acetate (150 mmol/L) was administered in drink to mice every day from day 0 of the food allergy model and occurrence of allergic diarrhea was analyzed to examine the effect of acetate on food allergy symptoms (n = approximately 5-10 mice per group). i.g., Intragastric; i.p., intraperitoneal. Journal of Allergy and Clinical Immunology 2016 137, 507-516.e8DOI: (10.1016/j.jaci.2015.08.016) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E6 ESBP attenuates food allergy in a dose-dependent manner. A, Purification of GST-tagged recombinant ESBP and ASBP. B and C, Injection of ESBP and ASBP into the mouse that was part of the food allergy model at various concentrations (0.5, 5, and 50 μg) and measurement of diarrhea occurrence (OVA + GST 50 μg vs OVA + ESBP 50 μg: **P < .01, n = 6 mice per group). Journal of Allergy and Clinical Immunology 2016 137, 507-516.e8DOI: (10.1016/j.jaci.2015.08.016) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E7 Effects of ESBP on mast cells, eosinophils, and Foxp3+ Treg cells in the small intestine. Mast cells, eosinophils, and Foxp3+ Treg cells in the LP were analyzed by using flow cytometry. A, Mast cells were gated as IgE+c-kit+ cells. B, Eosinophils were gated as Siglec-F+CCR3+ cells. C, Foxp3+ Treg cells were gated as CD4+Foxp3+ cells. Values represent means ± SEMs. Statistical analysis was performed by using 1-way ANOVA with the Newman-Keuls posttest. **P < .01. Data are representative of 2 independent experiments. Journal of Allergy and Clinical Immunology 2016 137, 507-516.e8DOI: (10.1016/j.jaci.2015.08.016) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E8 Administration of B longum KACC 91563 during the effector phase of the food allergy model has no suppressive effect. A, Experimental schedule for food allergy induction and B longum KACC 91563 administration in mice. B, Diarrhea occurrence (n = approximately 4-9 mice per group). C, Proportion of mast cells in the small intestine was determined by using fluorescence-activated cell sorting analysis (n = approximately 4-7 mice per group). D, MCPT-1 levels in serum. E, Paraffin-embedded sections of the small intestine were stained for mast cells. Numbers of mast cells were counted in 3 different fields (n = approximately 3-4 mice per group; original magnification ×400). Values represent means ± SEMs. i.g., Intragastric; i.p., intraperitoneal. Journal of Allergy and Clinical Immunology 2016 137, 507-516.e8DOI: (10.1016/j.jaci.2015.08.016) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions