HLA-DRB1 alleles control allergic bronchopulmonary aspergillosis–like pulmonary responses in humanized transgenic mice Sherri Koehm, BSc, Raymond G.

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Presentation transcript:

HLA-DRB1 alleles control allergic bronchopulmonary aspergillosis–like pulmonary responses in humanized transgenic mice Sherri Koehm, BSc, Raymond G. Slavin, MD, Patricia S. Hutcheson, BA, Theodore Trejo, BSc, Chella S. David, PhD, Clifford J. Bellone, PhD Journal of Allergy and Clinical Immunology Volume 120, Issue 3, Pages 570-577 (September 2007) DOI: 10.1016/j.jaci.2007.04.037 Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 Mice bearing susceptible DRB1∗1503 or resistance DRB1∗1502 alleles respond differently to A fumigatus. Mice were sensitized with Aspergillus species antigens and challenged intratracheally with Aspergillus species conidia. Seven or 14 days later, mice were killed, and the lungs were perfused with 10% formalin, tissue was embedded in paraffin, and 4- to 6-μm sections were stained with hematoxylin and eosin. Journal of Allergy and Clinical Immunology 2007 120, 570-577DOI: (10.1016/j.jaci.2007.04.037) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 Susceptible DRB1∗1503 mice respond prominently with eosinophil infiltration, and resistant DRB1∗1502 mice respond principally with T cells. Serial sections prepared from embedded lungs seen in Fig 1 were stained with anti-CD3 or monoclonal anti-BM8+ (an F4/80 epitope found on eosinophils and macrophage/monocytes). Sections were developed with peroxidase-conjugated anti-Ig antibodies and 3,3′-diaminobenzidine substrate and counterstained with hematoxylin. Journal of Allergy and Clinical Immunology 2007 120, 570-577DOI: (10.1016/j.jaci.2007.04.037) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 3 DRB1∗1503 mice display prominent bronchiolar mucin-producing goblet cells. Sections were stained with Masson trichome reagent displaying periodic acid-Schiff–positive mucin-producing goblet cells. Sections were counterstained with hematoxylin. Journal of Allergy and Clinical Immunology 2007 120, 570-577DOI: (10.1016/j.jaci.2007.04.037) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 4 Resistant DRB1∗1502 mice clear tracheally instilled conidia from the lung parenchyma. Paraffin tissue sections from lungs of both DRB1∗1503 and DRB1∗1502 mice were prepared and stained with Gomori methanamine silver reagent to visualize conidia within lung parenchyma. Journal of Allergy and Clinical Immunology 2007 120, 570-577DOI: (10.1016/j.jaci.2007.04.037) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 5 T cells from immune DRB1∗1503, DRB1∗1501, and DRB1∗1502 mice show robust antigen-induced proliferative responses. The illustration shows the stimulation index, wherein average background versus stimulated counts (tritiated thymidine incorporation; see the Methods section) for all strains were 4480 versus 19,470 cpm, respectively, for MLN cells, and 957 vs 6432 cpm, respectively, for splenocytes. Results are representative of 2 separate experiments, with 4 to 6 mice per strain per experiment. Journal of Allergy and Clinical Immunology 2007 120, 570-577DOI: (10.1016/j.jaci.2007.04.037) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 6 Total serum IgE levels and BAL cellular content are comparable in the resistant and susceptible strains. A, Sera were collected from normal unsensitized, sensitized only, and sensitized plus conidia instillation (14 days after instillation) groups. Levels were determined by means of ELISA, and average levels were determined from 10 to 15 mice per strain. B, BAL cell counts were determined morphologically from Wright-Giemsa–stained cytospin preparations. Average cell numbers were determined from 10-15 mice per strain. Journal of Allergy and Clinical Immunology 2007 120, 570-577DOI: (10.1016/j.jaci.2007.04.037) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions