Low-affinity allergen-specific IgE in cord blood and affinity maturation after birth Norio Kamemura, PhD, Norio Kawamoto, MD, PhD, Ryosuke Nakamura, PhD,

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Presentation transcript:

Low-affinity allergen-specific IgE in cord blood and affinity maturation after birth Norio Kamemura, PhD, Norio Kawamoto, MD, PhD, Ryosuke Nakamura, PhD, Reiko Teshima, PhD, Toshiyuki Fukao, MD, PhD, Hiroshi Kido, MD, PhD Journal of Allergy and Clinical Immunology Volume 133, Issue 3, Pages 904-905.e6 (March 2014) DOI: 10.1016/j.jaci.2013.09.034 Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 Competitive inhibition by OVM of the binding of OVM-specific IgE on the DLC-coated chip. CB and peripheral blood samples from 6- and 14-month-old infants (n = 9) were preincubated for 30 minutes with 0 to 1000 × 10-9 mol/L OVM and then the IgE bound on the chip was analyzed (A). Control binding without OVM was considered 100%. The % inhibition against OVM and OVM required for 50% inhibition (inset) as a measure of affinity were determined (B). Journal of Allergy and Clinical Immunology 2014 133, 904-905.e6DOI: (10.1016/j.jaci.2013.09.034) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 IgE-mediated human FcεRI cross-linking–induced luciferase expression in RS-ATL8 cells. Cells were preincubated with 100 BUe/mL OVM-specific IgE from CB and peripheral blood samples from 6- and 14-month-old infants. After washing, the cells were stimulated with 1 × 10-9 mol/L OVM and the induced luciferase was measured. Data are presented as fold increase with nonstimulated control considered as 1.0. *P < .05 and **P < .01. Journal of Allergy and Clinical Immunology 2014 133, 904-905.e6DOI: (10.1016/j.jaci.2013.09.034) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E1 Flow diagram of the specimen selection process. Journal of Allergy and Clinical Immunology 2014 133, 904-905.e6DOI: (10.1016/j.jaci.2013.09.034) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E2 OVM-specific IgE binding on the DLC-coated chip without cross-reaction with IgM, IgG, IgA, and IgD. Layout of various human immunoglobulins immobilized on a DLC-coated chip (100 and 200 × 10-9 mol/L) and rainbow display after reaction with fluorescence-labeled anti-IgE secondary antibody (A). Fluorescence value of each spot (B). Journal of Allergy and Clinical Immunology 2014 133, 904-905.e6DOI: (10.1016/j.jaci.2013.09.034) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E3 Effect of 0 to 40 × 10-3 mol/L DEA on the binding of serially diluted sera of CB and peripheral blood samples from 6- and 14-month-old infants to the OVM-immobilized DLC-coated chip. The curves of the binding of OVM-specific IgE to the chip in the presence of DEA (10-40 × 10-3 mol/L) were obtained by serial dilutions of CB (A) and blood samples from 14-month-old infants (n = 9, each) (B). Fluorescence without DEA for each dilution was considered 100%. The leftward shift by 40 × 10-3 mol/L DEA was measured at 50% of maximum fluorescence, and the results were expressed as log10 of this shift (C). ***P < .001. Journal of Allergy and Clinical Immunology 2014 133, 904-905.e6DOI: (10.1016/j.jaci.2013.09.034) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E4 Evaluation of anti–OVM-specific IgE-mediated FcεRI cross-linking in CB and peripheral blood samples obtained after birth in a luciferase-reporting mast cell line. OVM-specific IgE in CB and blood samples from 6- and 14-month-old infants were adjusted at 100 BUe/mL and then preincubated overnight with RS-ATL8 cells. After washing, the cells were induced with 0 to 10 × 10-9 mol/L OVM for 3 hours at 37°C. IgE-mediated FcεRI cross-linking–induced luciferase expression is presented as fold increase, with nonstimulated control as 1.0. Journal of Allergy and Clinical Immunology 2014 133, 904-905.e6DOI: (10.1016/j.jaci.2013.09.034) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions