Biomarkers for non-human primate Type-I hypersensitivity: Antigen-specific immunoglobulin E assays
| Autor: | Carla Forte, Darcey Clark, Padma K. Narayanan, Daniel T. Mytych, M. Benjamin Hock, Faith Shiota |
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| Rok vydání: | 2013 |
| Předmět: |
Hypersensitivity
Immediate Molecular Sequence Data Immunology Antigens CD34 Cross Reactions Biology Immunoglobulin E Epitope Epitopes chemistry.chemical_compound Immune system Antigen Hypersensitivity medicine Animals Humans Immunology and Allergy Amino Acid Sequence Mast Cells Immunoassay medicine.diagnostic_test Receptors IgE medicine.disease Molecular biology Macaca fascicularis chemistry Immunoglobulin G biology.protein Antibody Biomarkers Histamine Type I hypersensitivity |
| Zdroj: | Journal of Immunological Methods. 392:29-37 |
| ISSN: | 0022-1759 |
| DOI: | 10.1016/j.jim.2013.03.007 |
| Popis: | Immunoglobulin E (IgE) is the least abundant immunoglobulin in serum. However, development of an IgE immune response can induce IgE receptor-expressing cells to carry out potent effector functions. A reliable antigen-specific IgE biomarker method for use in non-human primate studies would facilitate (i) confirmation of Type-I hypersensitivity reactions during safety toxicology testing, and (ii) a better understanding of non-human primate models of allergic disease. We cloned and expressed a recombinant cynomolgus monkey IgE molecule in order to screen a panel of commercially available detection reagents raised against human IgE for cross-reactivity. The reagent most reactive to cynomolgus IgE was confirmed to be specific for IgE and did not bind recombinant cynomolgus monkey IgG1-4. A drug-specific IgE assay was developed on the MSD electrochemiluminescent (ECL) platform. The assay is capable of detecting 10 ng/mL drug-specific IgE. Importantly, the assay is able to detect IgE in the presence of excess IgG, the scenario likely to be present in a safety toxicology study. Using our ECL assay, we were able to confirm that serum from cynomolgus monkeys that had experienced clinical symptoms consistent with hypersensitivity responses contained IgE specific for a candidate therapeutic antibody. In addition, a bioassay for mast cell activation was developed using CD34(+)-derived cynomolgus monkey mast cells. This assay confirmed that plasma from animals identified as positive in the drug-specific IgE immunoassay contained biologically active IgE (i.e. could sensitize cultured mast cells), resulting in histamine release after exposure to the therapeutic antibody. These sensitive assays for Type-I hypersensitivity in the NHP can confirm that secondary events are downstream of immunogenicity. |
| Databáze: | OpenAIRE |
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