Staphylococcal protein A–formulated immune complexes suppress enterotoxin-induced cellular responses in nasal polyps Mitsuhiro Okano, MD, Tazuko Fujiwara,

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Presentation transcript:

Staphylococcal protein A–formulated immune complexes suppress enterotoxin-induced cellular responses in nasal polyps Mitsuhiro Okano, MD, Tazuko Fujiwara, BS, Shin Kariya, MD, Takenori Haruna, MD, Takaya Higaki, MD, Yasuyuki Noyama, MD, Sei- ichiro Makihara, MD, Kengo Kanai, MD, Kazunori Nishizaki, MD Journal of Allergy and Clinical Immunology Volume 136, Issue 2, Pages 343-350.e8 (August 2015) DOI: 10.1016/j.jaci.2014.10.058 Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 SpA-induced IL-5 (A), IL-13 (B), IFN-γ (C), IL-17A (D), and IL-10 (E) production by nasal polyp cells. Within the box plots, the box represents the 25th to 75th percentile range, the horizontal line indicates the median, and the vertical line indicates the 10th to 90th percentile range. P values were determined by using the Dunn test. ‡P < .001. Journal of Allergy and Clinical Immunology 2015 136, 343-350.e8DOI: (10.1016/j.jaci.2014.10.058) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 Cellular source of SpA-induced IL-10 production in nasal polyp cells. Nonadherent and adherent cells were cultured with or without 10 μg/mL SpA for 24 hours, and IL-10 levels in the supernatants were measured. P values were determined by using the Wilcoxon signed-rank test. Journal of Allergy and Clinical Immunology 2015 136, 343-350.e8DOI: (10.1016/j.jaci.2014.10.058) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 3 Enhanced IL-10 production by cells cultured with SpA plus IgG. DNPCs (A) and peripheral blood monocytes (B) were stimulated with SpA (10 μg/mL) in the presence or absence of IgG (100 μg/mL) for 24 hours, and IL-10 levels in the supernatants were measured. P values were determined by using the Dunn test. Journal of Allergy and Clinical Immunology 2015 136, 343-350.e8DOI: (10.1016/j.jaci.2014.10.058) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 4 Effect of SpA-IgG ICs on SEB-induced IL-5 (A), IL-13 (B), IFN-γ (C), and IL-17A (D) production by nasal polyp cells. DNPCs were treated with SpA (10 μg/mL) in the presence or absence of IgG (100 μg/mL) and then stimulated with SEB (1 ng/mL) for 72 hours. P values were determined by using the Wilcoxon signed-rank test. Journal of Allergy and Clinical Immunology 2015 136, 343-350.e8DOI: (10.1016/j.jaci.2014.10.058) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 5 Effect of SpA plus autologous serum on SEB-induced IL-5 (A), IL-13 (B), IFN-γ (C), and IL-17A (D) production by nasal polyp cells. DNPCs were treated with SpA (10 μg/mL) in the presence of 0%, 0.2%, 1%, or 5% autologous serum (AS) and then stimulated with SEB (1 ng/mL) for 72 hours. P values were determined by using the Wilcoxon signed-rank test. Journal of Allergy and Clinical Immunology 2015 136, 343-350.e8DOI: (10.1016/j.jaci.2014.10.058) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E1 IL-10 production by dispersed uncinate tissue cells (DUTCs). Cells were stimulated with or without SpA (10 μg/mL) for 24 hours, and IL-10 levels in the supernatant were measured. P values were determined by using the Wilcoxon signed-rank test. Journal of Allergy and Clinical Immunology 2015 136, 343-350.e8DOI: (10.1016/j.jaci.2014.10.058) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E2 Formation of SpA and IgG ICs. SpA (1 μg) and human IgG (11 μg) were mixed for 1 minute, 30 minutes, 1 hour, or 4 hours and then subjected to 4-15% polyacrylamide gel electrophoresis under native conditions. After electrophoresis, the gel was stained with Coomassie brilliant blue. Lanes containing only SpA and IgG are also shown as controls. Journal of Allergy and Clinical Immunology 2015 136, 343-350.e8DOI: (10.1016/j.jaci.2014.10.058) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E3 Effect of complement on SpA-IgG IC–induced IL-10 and IL-12 production by monocytes. Monocytes were cultured with the SpA and IgG mixture in the presence or absence of complement for 24 hours, and IL-10 (A) and IL-12 (B) levels in the supernatant were measured. P values were determined by using the Wilcoxon signed-rank test. Journal of Allergy and Clinical Immunology 2015 136, 343-350.e8DOI: (10.1016/j.jaci.2014.10.058) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E4 Effect of SpA-IgG ICs on IL-10 production by adherent cells in nasal polyps. Adherent DNPCs were cultured with or without the mixture of SpA (10 μg/mL) and IgG (100 μg/mL) for 24 hours, and IL-10 levels in the supernatant were measured. P values were determined by using the Wilcoxon signed-rank test. Journal of Allergy and Clinical Immunology 2015 136, 343-350.e8DOI: (10.1016/j.jaci.2014.10.058) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E5 Effect of IL-10 neutralization on SpA-IgG IC–induced suppression against SEB-induced IL-5 (A), IL-13 (B), IFN-γ (C), and IL-17A (D) production by nasal polyp cells. DNPCs were treated with or without the mixture of SpA (10 μg/mL) and IgG (100 μg/mL) and then stimulated with SEB (1 ng/mL) in the presence or absence of anti-human IgG mAb or control rat IgG1 (20 μg/mL) for 72 hours. P values were determined by using the Wilcoxon signed-rank test. Journal of Allergy and Clinical Immunology 2015 136, 343-350.e8DOI: (10.1016/j.jaci.2014.10.058) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E6 Effect of SpA-IgA ICs on SEB-induced IL-5 (A), IL-13 (B), IFN-γ (C), and IL-17A (D) production by nasal polyp cells. DNPCs were treated with SpA (10 μg/mL) in the presence or absence of IgA (100 μg/mL) and then stimulated with SEB (1 ng/mL) for 72 hours. P values were determined by using the Wilcoxon signed-rank test. Journal of Allergy and Clinical Immunology 2015 136, 343-350.e8DOI: (10.1016/j.jaci.2014.10.058) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E7 Enhanced IL-10 production by nasal polyp cells cultured with SpA plus autologous serum (AS). DNPCs were stimulated with SpA (10 μg/mL) in the presence of 0%, 0.2%, 1%, or 5% autologous serum for 24 hours, and IL-10 levels in the supernatant were measured. P values were determined by using the Wilcoxon signed-rank test. Journal of Allergy and Clinical Immunology 2015 136, 343-350.e8DOI: (10.1016/j.jaci.2014.10.058) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E8 Effect of SpA plus native or heat-inactivated autologous serum (AS) on SEB-induced IL-5 (A), IL-13 (B), IFN-γ (C), and IL-17A (D) production by nasal polyp cells. DNPCs were treated with SpA (10 μg/mL) in the presence or absence of 5% autologous serum with or without heat inactivation, and the cells were then stimulated with SEB (1 ng/mL) for 72 hours. P values were determined by using the Wilcoxon signed-rank test. Journal of Allergy and Clinical Immunology 2015 136, 343-350.e8DOI: (10.1016/j.jaci.2014.10.058) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions