Isocyanate vapor-induced antigenicity of human albumin

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Presentation transcript:

Isocyanate vapor-induced antigenicity of human albumin Adam V. Wisnewski, PhD, Meredith H. Stowe, PhD, André Cartier, MD, Qing Liu, PhD, Jian Liu, MS, Liang Chen, PhD, Carrie A. Redlich, MD, MPH Journal of Allergy and Clinical Immunology Volume 113, Issue 6, Pages 1178-1184 (June 2004) DOI: 10.1016/j.jaci.2004.03.009 Copyright © 2004 American Academy of Allergy, Asthma and Immunology Terms and Conditions

Fig 1 HDI vapors conjugate to albumin in fluid phase. Albumin in PBS was exposed to HDI vapors (≥100 ppb) and Western blotted with HDI-specific polyclonal rabbit antisera. Lane 1, unexposed; lane 2, 10 minutes; lane 3, 1 hour; lane 4, 4 hours; lane 5, 24 hours. Journal of Allergy and Clinical Immunology 2004 113, 1178-1184DOI: (10.1016/j.jaci.2004.03.009) Copyright © 2004 American Academy of Allergy, Asthma and Immunology Terms and Conditions

Fig 2 Conformational and/or charge changes in albumin after HDI vapor exposure. Native gel analysis of albumin exposed to HDI vapors for 24 hours (left) or varying concentrations of liquid HDI for 2 hours (right). Liquid exposure doses are expressed as a percentage based on volume. HDI substitution ratios (mol HDI:mol albumin), when detectable, are noted at the bottom. Data shown are representative of 3 different experiments. Journal of Allergy and Clinical Immunology 2004 113, 1178-1184DOI: (10.1016/j.jaci.2004.03.009) Copyright © 2004 American Academy of Allergy, Asthma and Immunology Terms and Conditions

Fig 3 Identification of HDI conjugation sites on albumin by MALDI-MS. Representative portions of the MALDI-MS mass/charge spectra comparing HDI- (top) and mock- (bottom) exposed albumin. Predominant new masses of 1317 Da and 1808 Da, consistently noted in the HDI-exposed albumin samples, are highlighted by the arrows. Journal of Allergy and Clinical Immunology 2004 113, 1178-1184DOI: (10.1016/j.jaci.2004.03.009) Copyright © 2004 American Academy of Allergy, Asthma and Immunology Terms and Conditions

Fig 4 Model depicting predominate HDI conjugation sites on human albumin. Two predominant loci of HDI conjugation to human albumin are predicted based on MALDI-MS/nanospray-MS analysis. Amino acids identified as predominate HDI conjugation sites (Lys414 and His247) are highlighted in yellow, HDI is depicted as a red star, and disulfide bonds between cysteines are drawn in orange. The 3 major domains of human albumin are color-coded purple (domain 1), blue (domain 2), and brown or gray (domain 3). Journal of Allergy and Clinical Immunology 2004 113, 1178-1184DOI: (10.1016/j.jaci.2004.03.009) Copyright © 2004 American Academy of Allergy, Asthma and Immunology Terms and Conditions

Fig 5 Specificity of human humoral response to HDI and dependency on albumin as the carrier. Unexposed and HDI vapor exposed human albumin (lanes 1 and 2, respectively), transferrin (lanes 3 and 4), lactoferrin (lanes 5 and 6), and bovine serum albumin (lanes 7 and 8) were Western blotted with (A) serum from a representative HDI-exposed subject, (B) HDI-specific polyclonal rabbit serum, or (C) pooled human serum from non–HDI-exposed subjects. Journal of Allergy and Clinical Immunology 2004 113, 1178-1184DOI: (10.1016/j.jaci.2004.03.009) Copyright © 2004 American Academy of Allergy, Asthma and Immunology Terms and Conditions

Fig 6 Association of vapor HDI-exposed albumin-specific IgG titers with exposure and asthma. Specific serum IgG titers versus (□) liquid or (▪) vapor HDI-exposed albumin conjugates (with substitution ratios of 37:1 and 3:1, respectively) were measured for healthy subjects, classified according to job category, and isocyanate asthmatics. Statistical significance of associations between titer and different job categories or isocyanate asthma is indicated. HDI-specific serum IgG was undetectable in all unexposed subjects tested to date (not shown). Journal of Allergy and Clinical Immunology 2004 113, 1178-1184DOI: (10.1016/j.jaci.2004.03.009) Copyright © 2004 American Academy of Allergy, Asthma and Immunology Terms and Conditions