Toll-like receptor 2 ligands activate human basophils for both IgE-dependent and IgE- independent secretion Anja P. Bieneman, BS, Kristin L. Chichester,

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Presentation transcript:

Toll-like receptor 2 ligands activate human basophils for both IgE-dependent and IgE- independent secretion Anja P. Bieneman, BS, Kristin L. Chichester, MS, Yi-Hsing Chen, MD, John T. Schroeder, PhD Journal of Allergy and Clinical Immunology Volume 115, Issue 2, Pages 295-301 (February 2005) DOI: 10.1016/j.jaci.2004.10.018 Copyright © 2005 American Academy of Allergy, Asthma and Immunology Terms and Conditions

Fig 1 Expression of TLR2 and TLR4 by human basophils (>98% purity) isolated from blood. A, RT-PCR results with TLR2 and TLR4 compared with the housekeeping gene, HPRT. B, Representative flow cytometry showing net mean fluorescence intensity (nMFI) comparing TLR2 and TLR4 (shaded) vs isotype control (nonshaded). PE, Phycoerythrin. Journal of Allergy and Clinical Immunology 2005 115, 295-301DOI: (10.1016/j.jaci.2004.10.018) Copyright © 2005 American Academy of Allergy, Asthma and Immunology Terms and Conditions

Fig 2 A, NFκB (p65 subunit) expressed in nuclear extracts of basophils treated 1 hour with medium alone (med.), peptidoglycan, LPS (10 μg/mL each), or phorbol 12-myristate 13-acetate (10 ng/mL)/ionomycin (500 ng/mL; P/I). B, Densitometry (average ± SEM; n=3) of these responses, normalized as a percentage of the P/I positive control. PGN, Peptidoglycan. Journal of Allergy and Clinical Immunology 2005 115, 295-301DOI: (10.1016/j.jaci.2004.10.018) Copyright © 2005 American Academy of Allergy, Asthma and Immunology Terms and Conditions

Fig 3 A-C, Mediator release and cytokine responses to peptidoglycan (PGN) or LPS alone (average ± SEM; n=6). Dotted line indicates level of detection. D-F, Preincubation (15 minutes) with PGN and LPS on anti-IgE–induced responses. Percent (average ± SEM; n=6) of anti-IgE control: 27% ± 8% (histamine), 260 pg/106 basophils (LTC4), and 203 ± 71 pg/106 basophils (IL-4). ∗P < .05 vs medium or control. Journal of Allergy and Clinical Immunology 2005 115, 295-301DOI: (10.1016/j.jaci.2004.10.018) Copyright © 2005 American Academy of Allergy, Asthma and Immunology Terms and Conditions

Fig 4 A, Basophil IL-13 (average ± SEM; n=10). Line denotes level of detection. B, Percent enhancement (average ± SEM; n=7) of IL-13 induced with 10 ng/mL IL-3 (control levels, 128 ± 45 pg/106 basophils). C, Peptidoglycan (PGN; 10 μg/mL) or medium (control) on the IL-13 secreted by basophils of allergic (A) versus nonallergic (NA) subjects. Average ± SEM (n=4) for each group. ∗P < .05; **P < .01 versus all other conditions. Journal of Allergy and Clinical Immunology 2005 115, 295-301DOI: (10.1016/j.jaci.2004.10.018) Copyright © 2005 American Academy of Allergy, Asthma and Immunology Terms and Conditions

Fig 5 A, IL-13 secretion with the indicated TLR2 ligands combined with IL-3 (average ± SEM; n=3). B, IL-4 in response to 15-minute preincubation with TLR2 ligands followed by 3 hours with anti-IgE (average ± SEM; n=4). Peptidoglycan (PGN) was 10 μg/mL. ∗P < .05 vs anti-IgE or IL-3 alone. Journal of Allergy and Clinical Immunology 2005 115, 295-301DOI: (10.1016/j.jaci.2004.10.018) Copyright © 2005 American Academy of Allergy, Asthma and Immunology Terms and Conditions

Fig 6 A, NFκB p65 induced by peptidoglycan (PGN; 10 μg/mL) and its inhibition by SN50 (10 μg/mL). B and C, Effect of SN50 or control peptide (SN50-M) on priming of IgE-mediated IL-4 (average ± SEM; n=4) and of IL-13 induced by IL-3 (5 ng/mL), respectively. TLR2 ligands at 10 μg/mL. ∗P < .05 vs anti-IgE; **P < .05 vs other conditions. EXP, Experiment; med., medium alone. Journal of Allergy and Clinical Immunology 2005 115, 295-301DOI: (10.1016/j.jaci.2004.10.018) Copyright © 2005 American Academy of Allergy, Asthma and Immunology Terms and Conditions