Secretory IgA induces antigen-independent eosinophil survival and cytokine production without inducing effector functions Kathleen R. Bartemes, BA, Kate.

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Presentation transcript:

Secretory IgA induces antigen-independent eosinophil survival and cytokine production without inducing effector functions Kathleen R. Bartemes, BA, Kate M. Cooper, BA, Kerry L. Drain, MD, Hirohito Kita, MD Journal of Allergy and Clinical Immunology Volume 116, Issue 4, Pages 827-835 (October 2005) DOI: 10.1016/j.jaci.2005.07.014 Copyright © 2005 American Academy of Allergy, Asthma and Immunology Terms and Conditions

Fig 1 Superoxide production and degranulation from eosinophils stimulated by S-IgA. Eosinophils were incubated for 3 hours with soluble or immobilized S-IgA (see Methods). A, Superoxide production was measured by cytochrome c reduction. B, EDN released into the supernatants was measured by RIA. Results are means ± SEMs, n = 3. ∗P < .05; ∗∗P < .01 compared with medium. Journal of Allergy and Clinical Immunology 2005 116, 827-835DOI: (10.1016/j.jaci.2005.07.014) Copyright © 2005 American Academy of Allergy, Asthma and Immunology Terms and Conditions

Fig 2 Eosinophil viability with S-IgA and IgG. A, Eosinophils were incubated with soluble S-IgA (10 μg/mL), immobilized S-IgA (wells precoated with 10 μg/mL S-IgA), IL-5 (100 pg/mL), or medium for 24 to 96 hours. B, Eosinophils were incubated for 96 hours with soluble S-IgA or IgG; viability was determined by PI and flow cytometry. Results are mean percent of viable cells ± SEMs, n = 3 (A) or 4 (B) experiments. ∗P < .05; ∗∗P < .01 compared with medium. Journal of Allergy and Clinical Immunology 2005 116, 827-835DOI: (10.1016/j.jaci.2005.07.014) Copyright © 2005 American Academy of Allergy, Asthma and Immunology Terms and Conditions

Fig 3 S-IgA fractionation and eosinophil viability. A, S-IgA was fractionated by using a size exclusion column. Molecular mass markers over the same column eluted at the numbered fraction. B, Alternate column fractions analyzed by nondenaturing, nonreducing PAGE; the gel was silver-stained. C, Eosinophils were incubated for 96 hours with 10 μg/mL unfractionated (S-IgA) or fractionated (Fr. 14) soluble S-IgA; see Fig 2 for viability. Journal of Allergy and Clinical Immunology 2005 116, 827-835DOI: (10.1016/j.jaci.2005.07.014) Copyright © 2005 American Academy of Allergy, Asthma and Immunology Terms and Conditions

Fig 4 Ligation of FcαRI (CD89) by soluble mAb or soluble ligand and eosinophil survival. Eosinophils were incubated for 96 hours with (A) serum IgA or (B) anti-FcαRI mAb or mouse IgG1 control. See Fig 2 for viability. Results are mean percent of viable cells ± SEMs, n = 4 (A) or 5 (B). ∗P < .05 compared with medium (A) or corresponding concentration of control antibody (B). Journal of Allergy and Clinical Immunology 2005 116, 827-835DOI: (10.1016/j.jaci.2005.07.014) Copyright © 2005 American Academy of Allergy, Asthma and Immunology Terms and Conditions

Fig 5 Autocrine cytokines and eosinophil survival induced by soluble S-IgA. A, Eosinophils were incubated for 96 hours with soluble S-IgA (10 μg/mL) with or without polyclonal antibody (anti–IL-5, anti–IL-3, or anti–GM-CSF). Control cells were cultured in medium (None). See Fig 2 for viability. Results are mean percent of viable cells + SEMs, n = 3. ∗P < .05 compared with no antibody. B, Eosinophils were incubated 16 hours with medium or 10 μg/mL of soluble S-IgA or IgG. Supernatant GM-CSF was measured by ELISA. Results are means + SEMs, n = 3. ∗∗P < .01 compared with medium. Journal of Allergy and Clinical Immunology 2005 116, 827-835DOI: (10.1016/j.jaci.2005.07.014) Copyright © 2005 American Academy of Allergy, Asthma and Immunology Terms and Conditions

Fig 6 Effects of soluble S-IgA and IgA on the ERK kinase pathway. A, Eosinophils were incubated for 5 or 30 minutes with 10 μg/mL soluble S-IgA or IgA. Phosphorylation of ERK1/2 in lysates was detected by Western blot (antiphosphorylated p44/42, upper panel; anti-p44/42, lower panel). B, Eosinophils were incubated 16 hours with medium or 10 μg/mL soluble S-IgA or IgA with or without increasing PD98059 concentrations. Supernatant IL-8 or GM-CSF levels were measured by ELISA. Results are means + SEMs, n = 3. ∗P < .05; ∗∗P < .01 compared with no PD98059. Journal of Allergy and Clinical Immunology 2005 116, 827-835DOI: (10.1016/j.jaci.2005.07.014) Copyright © 2005 American Academy of Allergy, Asthma and Immunology Terms and Conditions