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

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Presentation transcript:

Induction of long-lived allergen-specific plasma cells by mucosal allergen challenge Elke O. Luger, PhD, Verena Fokuhl, MSc, Michael Wegmann, PhD, Melanie Abram, MSc, Kati Tillack, MSc, Gernot Achatz, PhD, Rudolf A. Manz, PhD, Margitta Worm, MD, Andreas Radbruch, PhD, Harald Renz, MD Journal of Allergy and Clinical Immunology Volume 124, Issue 4, Pages 819-826.e4 (October 2009) DOI: 10.1016/j.jaci.2009.06.047 Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 Cyclophosphamide (CyP)-resistant ovalbumin (OVA)-specific PCs in bone marrow and spleen. A, Experimental setup. B, Survival of OVA-specific immunoglobulin ASCs detected by ELISpot at day 100. C, Specific effect of CyP shown by reduction of ASCs at day 70. Each symbol represents the number of ASCs per organ per single mouse. Data from 2 independent experiments, 16 mice each. i.p., Intraperitoneal. Journal of Allergy and Clinical Immunology 2009 124, 819-826.e4DOI: (10.1016/j.jaci.2009.06.047) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 Cyclophosphamide (CyP)-resistant ovalbumin (OVA)-specific PCs in bone marrow and spleen. A, Experimental setup. B, Survival of OVA-specific immunoglobulin ASCs detected by ELISpot at day 100. C, Specific effect of CyP shown by reduction of ASCs at day 70. Each symbol represents the number of ASCs per organ per single mouse. Data from 2 independent experiments, 16 mice each. i.p., Intraperitoneal. Journal of Allergy and Clinical Immunology 2009 124, 819-826.e4DOI: (10.1016/j.jaci.2009.06.047) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 Cyclophosphamide (CyP)-resistant ovalbumin (OVA)-specific PCs in bone marrow and spleen. A, Experimental setup. B, Survival of OVA-specific immunoglobulin ASCs detected by ELISpot at day 100. C, Specific effect of CyP shown by reduction of ASCs at day 70. Each symbol represents the number of ASCs per organ per single mouse. Data from 2 independent experiments, 16 mice each. i.p., Intraperitoneal. Journal of Allergy and Clinical Immunology 2009 124, 819-826.e4DOI: (10.1016/j.jaci.2009.06.047) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 Cyclophosphamide (CyP)-resistant ovalbumin (OVA)-specific PCs in bone marrow and spleen. A, Experimental setup. B, Survival of OVA-specific immunoglobulin ASCs detected by ELISpot at day 100. C, Specific effect of CyP shown by reduction of ASCs at day 70. Each symbol represents the number of ASCs per organ per single mouse. Data from 2 independent experiments, 16 mice each. i.p., Intraperitoneal. Journal of Allergy and Clinical Immunology 2009 124, 819-826.e4DOI: (10.1016/j.jaci.2009.06.047) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 Cyclophosphamide (CyP)-resistant ovalbumin (OVA)-specific PCs in bone marrow and spleen. A, Experimental setup. B, Survival of OVA-specific immunoglobulin ASCs detected by ELISpot at day 100. C, Specific effect of CyP shown by reduction of ASCs at day 70. Each symbol represents the number of ASCs per organ per single mouse. Data from 2 independent experiments, 16 mice each. i.p., Intraperitoneal. Journal of Allergy and Clinical Immunology 2009 124, 819-826.e4DOI: (10.1016/j.jaci.2009.06.047) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 Cyclophosphamide (CyP)-resistant ovalbumin (OVA)-specific PCs in bone marrow and spleen. A, Experimental setup. B, Survival of OVA-specific immunoglobulin ASCs detected by ELISpot at day 100. C, Specific effect of CyP shown by reduction of ASCs at day 70. Each symbol represents the number of ASCs per organ per single mouse. Data from 2 independent experiments, 16 mice each. i.p., Intraperitoneal. Journal of Allergy and Clinical Immunology 2009 124, 819-826.e4DOI: (10.1016/j.jaci.2009.06.047) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 Cyclophosphamide (CyP)-resistant ovalbumin (OVA)-specific PCs in bone marrow and spleen. A, Experimental setup. B, Survival of OVA-specific immunoglobulin ASCs detected by ELISpot at day 100. C, Specific effect of CyP shown by reduction of ASCs at day 70. Each symbol represents the number of ASCs per organ per single mouse. Data from 2 independent experiments, 16 mice each. i.p., Intraperitoneal. Journal of Allergy and Clinical Immunology 2009 124, 819-826.e4DOI: (10.1016/j.jaci.2009.06.047) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 Immigration and kinetic of ovalbumin (OVA)-specific IgG1, IgA, and IgE ASCs. A, Experimental setup. B, Lung ELISpot of OVA-sensitized (●) and OVA-sensitized plus OVA aerosol–challenged mice (▴). C, Bone marrow/spleen ELISpot of OVA-sensitized (●) and sensitized plus OVA-challenged mice (▴). i.p., Intraperitoneal. Journal of Allergy and Clinical Immunology 2009 124, 819-826.e4DOI: (10.1016/j.jaci.2009.06.047) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 Immigration and kinetic of ovalbumin (OVA)-specific IgG1, IgA, and IgE ASCs. A, Experimental setup. B, Lung ELISpot of OVA-sensitized (●) and OVA-sensitized plus OVA aerosol–challenged mice (▴). C, Bone marrow/spleen ELISpot of OVA-sensitized (●) and sensitized plus OVA-challenged mice (▴). i.p., Intraperitoneal. Journal of Allergy and Clinical Immunology 2009 124, 819-826.e4DOI: (10.1016/j.jaci.2009.06.047) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 Immigration and kinetic of ovalbumin (OVA)-specific IgG1, IgA, and IgE ASCs. A, Experimental setup. B, Lung ELISpot of OVA-sensitized (●) and OVA-sensitized plus OVA aerosol–challenged mice (▴). C, Bone marrow/spleen ELISpot of OVA-sensitized (●) and sensitized plus OVA-challenged mice (▴). i.p., Intraperitoneal. Journal of Allergy and Clinical Immunology 2009 124, 819-826.e4DOI: (10.1016/j.jaci.2009.06.047) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 Immigration and kinetic of ovalbumin (OVA)-specific IgG1, IgA, and IgE ASCs. A, Experimental setup. B, Lung ELISpot of OVA-sensitized (●) and OVA-sensitized plus OVA aerosol–challenged mice (▴). C, Bone marrow/spleen ELISpot of OVA-sensitized (●) and sensitized plus OVA-challenged mice (▴). i.p., Intraperitoneal. Journal of Allergy and Clinical Immunology 2009 124, 819-826.e4DOI: (10.1016/j.jaci.2009.06.047) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 Immigration and kinetic of ovalbumin (OVA)-specific IgG1, IgA, and IgE ASCs. A, Experimental setup. B, Lung ELISpot of OVA-sensitized (●) and OVA-sensitized plus OVA aerosol–challenged mice (▴). C, Bone marrow/spleen ELISpot of OVA-sensitized (●) and sensitized plus OVA-challenged mice (▴). i.p., Intraperitoneal. Journal of Allergy and Clinical Immunology 2009 124, 819-826.e4DOI: (10.1016/j.jaci.2009.06.047) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 Immigration and kinetic of ovalbumin (OVA)-specific IgG1, IgA, and IgE ASCs. A, Experimental setup. B, Lung ELISpot of OVA-sensitized (●) and OVA-sensitized plus OVA aerosol–challenged mice (▴). C, Bone marrow/spleen ELISpot of OVA-sensitized (●) and sensitized plus OVA-challenged mice (▴). i.p., Intraperitoneal. Journal of Allergy and Clinical Immunology 2009 124, 819-826.e4DOI: (10.1016/j.jaci.2009.06.047) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 Immigration and kinetic of ovalbumin (OVA)-specific IgG1, IgA, and IgE ASCs. A, Experimental setup. B, Lung ELISpot of OVA-sensitized (●) and OVA-sensitized plus OVA aerosol–challenged mice (▴). C, Bone marrow/spleen ELISpot of OVA-sensitized (●) and sensitized plus OVA-challenged mice (▴). i.p., Intraperitoneal. Journal of Allergy and Clinical Immunology 2009 124, 819-826.e4DOI: (10.1016/j.jaci.2009.06.047) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 Immigration and kinetic of ovalbumin (OVA)-specific IgG1, IgA, and IgE ASCs. A, Experimental setup. B, Lung ELISpot of OVA-sensitized (●) and OVA-sensitized plus OVA aerosol–challenged mice (▴). C, Bone marrow/spleen ELISpot of OVA-sensitized (●) and sensitized plus OVA-challenged mice (▴). i.p., Intraperitoneal. Journal of Allergy and Clinical Immunology 2009 124, 819-826.e4DOI: (10.1016/j.jaci.2009.06.047) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 Immigration and kinetic of ovalbumin (OVA)-specific IgG1, IgA, and IgE ASCs. A, Experimental setup. B, Lung ELISpot of OVA-sensitized (●) and OVA-sensitized plus OVA aerosol–challenged mice (▴). C, Bone marrow/spleen ELISpot of OVA-sensitized (●) and sensitized plus OVA-challenged mice (▴). i.p., Intraperitoneal. Journal of Allergy and Clinical Immunology 2009 124, 819-826.e4DOI: (10.1016/j.jaci.2009.06.047) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 Immigration and kinetic of ovalbumin (OVA)-specific IgG1, IgA, and IgE ASCs. A, Experimental setup. B, Lung ELISpot of OVA-sensitized (●) and OVA-sensitized plus OVA aerosol–challenged mice (▴). C, Bone marrow/spleen ELISpot of OVA-sensitized (●) and sensitized plus OVA-challenged mice (▴). i.p., Intraperitoneal. Journal of Allergy and Clinical Immunology 2009 124, 819-826.e4DOI: (10.1016/j.jaci.2009.06.047) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 Immigration and kinetic of ovalbumin (OVA)-specific IgG1, IgA, and IgE ASCs. A, Experimental setup. B, Lung ELISpot of OVA-sensitized (●) and OVA-sensitized plus OVA aerosol–challenged mice (▴). C, Bone marrow/spleen ELISpot of OVA-sensitized (●) and sensitized plus OVA-challenged mice (▴). i.p., Intraperitoneal. Journal of Allergy and Clinical Immunology 2009 124, 819-826.e4DOI: (10.1016/j.jaci.2009.06.047) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 3 Ovalbumin (OVA) aerosol boosts systemic immune response. A, Experimental setup. B, Cryostaining of splenic IgE PCs with (□) and without (●) OVA inhalation. n = 6-16. Representative IgE PC staining. C, OVA-specific PCs in spleen with and without 2 weeks of OVA aerosol treatment. Gated on CD138. D, Increase of OVA-IgE ASCs in bone marrow during OVA aerosol by ELISpot. n = 6. i.p., Intraperitoneal. Journal of Allergy and Clinical Immunology 2009 124, 819-826.e4DOI: (10.1016/j.jaci.2009.06.047) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 3 Ovalbumin (OVA) aerosol boosts systemic immune response. A, Experimental setup. B, Cryostaining of splenic IgE PCs with (□) and without (●) OVA inhalation. n = 6-16. Representative IgE PC staining. C, OVA-specific PCs in spleen with and without 2 weeks of OVA aerosol treatment. Gated on CD138. D, Increase of OVA-IgE ASCs in bone marrow during OVA aerosol by ELISpot. n = 6. i.p., Intraperitoneal. Journal of Allergy and Clinical Immunology 2009 124, 819-826.e4DOI: (10.1016/j.jaci.2009.06.047) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 3 Ovalbumin (OVA) aerosol boosts systemic immune response. A, Experimental setup. B, Cryostaining of splenic IgE PCs with (□) and without (●) OVA inhalation. n = 6-16. Representative IgE PC staining. C, OVA-specific PCs in spleen with and without 2 weeks of OVA aerosol treatment. Gated on CD138. D, Increase of OVA-IgE ASCs in bone marrow during OVA aerosol by ELISpot. n = 6. i.p., Intraperitoneal. Journal of Allergy and Clinical Immunology 2009 124, 819-826.e4DOI: (10.1016/j.jaci.2009.06.047) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 3 Ovalbumin (OVA) aerosol boosts systemic immune response. A, Experimental setup. B, Cryostaining of splenic IgE PCs with (□) and without (●) OVA inhalation. n = 6-16. Representative IgE PC staining. C, OVA-specific PCs in spleen with and without 2 weeks of OVA aerosol treatment. Gated on CD138. D, Increase of OVA-IgE ASCs in bone marrow during OVA aerosol by ELISpot. n = 6. i.p., Intraperitoneal. Journal of Allergy and Clinical Immunology 2009 124, 819-826.e4DOI: (10.1016/j.jaci.2009.06.047) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 3 Ovalbumin (OVA) aerosol boosts systemic immune response. A, Experimental setup. B, Cryostaining of splenic IgE PCs with (□) and without (●) OVA inhalation. n = 6-16. Representative IgE PC staining. C, OVA-specific PCs in spleen with and without 2 weeks of OVA aerosol treatment. Gated on CD138. D, Increase of OVA-IgE ASCs in bone marrow during OVA aerosol by ELISpot. n = 6. i.p., Intraperitoneal. Journal of Allergy and Clinical Immunology 2009 124, 819-826.e4DOI: (10.1016/j.jaci.2009.06.047) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 4 Survival of long-lived, ovalbumin (OVA)-specific PCs at day 100 after aerosol challenge. A, Experimental setup. B, Long-lived OVA-specific ASCs at day 100 in bone marrow, spleen, and lung, detected by ELISpot. (●) Systemically immunized, (○) systemically immunized plus cyclophosphamide (CyP)-treated (days 74-77), (▴) systemically immunized plus aerosol-challenged, (Δ) systemically immunized, aerosol and cyclophosphamide–treated. n = 8 per group. i.p., Intraperitoneal. Journal of Allergy and Clinical Immunology 2009 124, 819-826.e4DOI: (10.1016/j.jaci.2009.06.047) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 4 Survival of long-lived, ovalbumin (OVA)-specific PCs at day 100 after aerosol challenge. A, Experimental setup. B, Long-lived OVA-specific ASCs at day 100 in bone marrow, spleen, and lung, detected by ELISpot. (●) Systemically immunized, (○) systemically immunized plus cyclophosphamide (CyP)-treated (days 74-77), (▴) systemically immunized plus aerosol-challenged, (Δ) systemically immunized, aerosol and cyclophosphamide–treated. n = 8 per group. i.p., Intraperitoneal. Journal of Allergy and Clinical Immunology 2009 124, 819-826.e4DOI: (10.1016/j.jaci.2009.06.047) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 4 Survival of long-lived, ovalbumin (OVA)-specific PCs at day 100 after aerosol challenge. A, Experimental setup. B, Long-lived OVA-specific ASCs at day 100 in bone marrow, spleen, and lung, detected by ELISpot. (●) Systemically immunized, (○) systemically immunized plus cyclophosphamide (CyP)-treated (days 74-77), (▴) systemically immunized plus aerosol-challenged, (Δ) systemically immunized, aerosol and cyclophosphamide–treated. n = 8 per group. i.p., Intraperitoneal. Journal of Allergy and Clinical Immunology 2009 124, 819-826.e4DOI: (10.1016/j.jaci.2009.06.047) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 4 Survival of long-lived, ovalbumin (OVA)-specific PCs at day 100 after aerosol challenge. A, Experimental setup. B, Long-lived OVA-specific ASCs at day 100 in bone marrow, spleen, and lung, detected by ELISpot. (●) Systemically immunized, (○) systemically immunized plus cyclophosphamide (CyP)-treated (days 74-77), (▴) systemically immunized plus aerosol-challenged, (Δ) systemically immunized, aerosol and cyclophosphamide–treated. n = 8 per group. i.p., Intraperitoneal. Journal of Allergy and Clinical Immunology 2009 124, 819-826.e4DOI: (10.1016/j.jaci.2009.06.047) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 4 Survival of long-lived, ovalbumin (OVA)-specific PCs at day 100 after aerosol challenge. A, Experimental setup. B, Long-lived OVA-specific ASCs at day 100 in bone marrow, spleen, and lung, detected by ELISpot. (●) Systemically immunized, (○) systemically immunized plus cyclophosphamide (CyP)-treated (days 74-77), (▴) systemically immunized plus aerosol-challenged, (Δ) systemically immunized, aerosol and cyclophosphamide–treated. n = 8 per group. i.p., Intraperitoneal. Journal of Allergy and Clinical Immunology 2009 124, 819-826.e4DOI: (10.1016/j.jaci.2009.06.047) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 4 Survival of long-lived, ovalbumin (OVA)-specific PCs at day 100 after aerosol challenge. A, Experimental setup. B, Long-lived OVA-specific ASCs at day 100 in bone marrow, spleen, and lung, detected by ELISpot. (●) Systemically immunized, (○) systemically immunized plus cyclophosphamide (CyP)-treated (days 74-77), (▴) systemically immunized plus aerosol-challenged, (Δ) systemically immunized, aerosol and cyclophosphamide–treated. n = 8 per group. i.p., Intraperitoneal. Journal of Allergy and Clinical Immunology 2009 124, 819-826.e4DOI: (10.1016/j.jaci.2009.06.047) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 4 Survival of long-lived, ovalbumin (OVA)-specific PCs at day 100 after aerosol challenge. A, Experimental setup. B, Long-lived OVA-specific ASCs at day 100 in bone marrow, spleen, and lung, detected by ELISpot. (●) Systemically immunized, (○) systemically immunized plus cyclophosphamide (CyP)-treated (days 74-77), (▴) systemically immunized plus aerosol-challenged, (Δ) systemically immunized, aerosol and cyclophosphamide–treated. n = 8 per group. i.p., Intraperitoneal. Journal of Allergy and Clinical Immunology 2009 124, 819-826.e4DOI: (10.1016/j.jaci.2009.06.047) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 4 Survival of long-lived, ovalbumin (OVA)-specific PCs at day 100 after aerosol challenge. A, Experimental setup. B, Long-lived OVA-specific ASCs at day 100 in bone marrow, spleen, and lung, detected by ELISpot. (●) Systemically immunized, (○) systemically immunized plus cyclophosphamide (CyP)-treated (days 74-77), (▴) systemically immunized plus aerosol-challenged, (Δ) systemically immunized, aerosol and cyclophosphamide–treated. n = 8 per group. i.p., Intraperitoneal. Journal of Allergy and Clinical Immunology 2009 124, 819-826.e4DOI: (10.1016/j.jaci.2009.06.047) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 4 Survival of long-lived, ovalbumin (OVA)-specific PCs at day 100 after aerosol challenge. A, Experimental setup. B, Long-lived OVA-specific ASCs at day 100 in bone marrow, spleen, and lung, detected by ELISpot. (●) Systemically immunized, (○) systemically immunized plus cyclophosphamide (CyP)-treated (days 74-77), (▴) systemically immunized plus aerosol-challenged, (Δ) systemically immunized, aerosol and cyclophosphamide–treated. n = 8 per group. i.p., Intraperitoneal. Journal of Allergy and Clinical Immunology 2009 124, 819-826.e4DOI: (10.1016/j.jaci.2009.06.047) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 4 Survival of long-lived, ovalbumin (OVA)-specific PCs at day 100 after aerosol challenge. A, Experimental setup. B, Long-lived OVA-specific ASCs at day 100 in bone marrow, spleen, and lung, detected by ELISpot. (●) Systemically immunized, (○) systemically immunized plus cyclophosphamide (CyP)-treated (days 74-77), (▴) systemically immunized plus aerosol-challenged, (Δ) systemically immunized, aerosol and cyclophosphamide–treated. n = 8 per group. i.p., Intraperitoneal. Journal of Allergy and Clinical Immunology 2009 124, 819-826.e4DOI: (10.1016/j.jaci.2009.06.047) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Cyclophosphamide -resistant ovalbumin-specific PCs in bone marrow and spleen. A, Survival of total immunoglobulin ASCs detected by ELISpot at day 100. B, Representative ELISpot. Journal of Allergy and Clinical Immunology 2009 124, 819-826.e4DOI: (10.1016/j.jaci.2009.06.047) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Cyclophosphamide -resistant ovalbumin-specific PCs in bone marrow and spleen. A, Survival of total immunoglobulin ASCs detected by ELISpot at day 100. B, Representative ELISpot. Journal of Allergy and Clinical Immunology 2009 124, 819-826.e4DOI: (10.1016/j.jaci.2009.06.047) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Immigration and kinetic of ovalbumin-specific IgG1, IgA, and IgE ASCs Immigration and kinetic of ovalbumin-specific IgG1, IgA, and IgE ASCs. A, Experimental setup. B, Cryostaining from day 29 of T cells (anti-CD3), B cells (anti-B220), BCR on the surface or in the cytoplasm of B cells/plasmablasts/PCs (anti-κ/λ staining), IgE-positive and IgG-positive B cells and PCs (anti-IgE/anti-IgG). Journal of Allergy and Clinical Immunology 2009 124, 819-826.e4DOI: (10.1016/j.jaci.2009.06.047) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Immigration and kinetic of ovalbumin-specific IgG1, IgA, and IgE ASCs Immigration and kinetic of ovalbumin-specific IgG1, IgA, and IgE ASCs. A, Experimental setup. B, Cryostaining from day 29 of T cells (anti-CD3), B cells (anti-B220), BCR on the surface or in the cytoplasm of B cells/plasmablasts/PCs (anti-κ/λ staining), IgE-positive and IgG-positive B cells and PCs (anti-IgE/anti-IgG). Journal of Allergy and Clinical Immunology 2009 124, 819-826.e4DOI: (10.1016/j.jaci.2009.06.047) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Immigration and kinetic of ovalbumin-specific IgG1, IgA, and IgE ASCs Immigration and kinetic of ovalbumin-specific IgG1, IgA, and IgE ASCs. A, Experimental setup. B, Cryostaining from day 29 of T cells (anti-CD3), B cells (anti-B220), BCR on the surface or in the cytoplasm of B cells/plasmablasts/PCs (anti-κ/λ staining), IgE-positive and IgG-positive B cells and PCs (anti-IgE/anti-IgG). Journal of Allergy and Clinical Immunology 2009 124, 819-826.e4DOI: (10.1016/j.jaci.2009.06.047) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Immigration and kinetic of ovalbumin-specific IgG1, IgA, and IgE ASCs Immigration and kinetic of ovalbumin-specific IgG1, IgA, and IgE ASCs. A, Experimental setup. B, Cryostaining from day 29 of T cells (anti-CD3), B cells (anti-B220), BCR on the surface or in the cytoplasm of B cells/plasmablasts/PCs (anti-κ/λ staining), IgE-positive and IgG-positive B cells and PCs (anti-IgE/anti-IgG). Journal of Allergy and Clinical Immunology 2009 124, 819-826.e4DOI: (10.1016/j.jaci.2009.06.047) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Immigration and kinetic of ovalbumin-specific IgG1, IgA, and IgE ASCs Immigration and kinetic of ovalbumin-specific IgG1, IgA, and IgE ASCs. A, Experimental setup. B, Cryostaining from day 29 of T cells (anti-CD3), B cells (anti-B220), BCR on the surface or in the cytoplasm of B cells/plasmablasts/PCs (anti-κ/λ staining), IgE-positive and IgG-positive B cells and PCs (anti-IgE/anti-IgG). Journal of Allergy and Clinical Immunology 2009 124, 819-826.e4DOI: (10.1016/j.jaci.2009.06.047) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Immigration and kinetic of ovalbumin-specific IgG1, IgA, and IgE ASCs Immigration and kinetic of ovalbumin-specific IgG1, IgA, and IgE ASCs. A, Experimental setup. B, Cryostaining from day 29 of T cells (anti-CD3), B cells (anti-B220), BCR on the surface or in the cytoplasm of B cells/plasmablasts/PCs (anti-κ/λ staining), IgE-positive and IgG-positive B cells and PCs (anti-IgE/anti-IgG). Journal of Allergy and Clinical Immunology 2009 124, 819-826.e4DOI: (10.1016/j.jaci.2009.06.047) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Ovalbumin aerosol boosts systemic immune response Ovalbumin aerosol boosts systemic immune response. Total and ovalbumin-specific ELISA. Data from 2 independent experiments. n = 12 each. Journal of Allergy and Clinical Immunology 2009 124, 819-826.e4DOI: (10.1016/j.jaci.2009.06.047) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Survival of long-lived, ovalbumin-specific PCs at day 100 after aerosol challenge. Ovalbumin-specific IgE and IgG1 ELISA Data from 2 independent experiments. n = 10. Journal of Allergy and Clinical Immunology 2009 124, 819-826.e4DOI: (10.1016/j.jaci.2009.06.047) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions