title: Optimization of the THP-1 activation assay to detect pharmaceuticals with potential to cause immune mediated drug reactions.
abstract: Despite important impacts of systemic hypersensitivity induced by pharmaceuticals, for such endpoint no reliable preclinical approaches are available. We previously established an in vitro test to identify contact and respiratory allergens based on interleukin-8 (IL-8) production in THP-1 cells. Here, we challenged it for identification of pharmaceuticals associated with systemic hypersensitivity reactions, with the idea that drug sensitizers share common mechanisms of cell activation. Cells were exposed to drugs associated with systemic hypersensitivity reactions (streptozotocin, sulfamethoxazole, neomycin, probenecid, clonidine, procainamide, ofloxacin, methyl salicylate), while metformin was used as negative drug. Differently to chemicals, drugs tested were well tolerated, except clonidine and probenecid, with no signs of cytotoxicity up to 1-2mg/ml. THP-1 activation assay was adjusted, and conditions, that allow identification of all sensitizing drugs tested, were established. Next, using streptozotocin and selective inhibitors of PKC-β and p38 MAPK, two pathways involved in chemical allergen-induced cell activation, we tested the hypothesis that similar pathways were also involved in drug-induced IL-8 production and CD86 upregulation. Results indicated that drugs and chemical allergens share similar activation pathways. Finally, we made a structure-activity hypothesis related to hypersensitivity reactions, trying to individuate structural requisite that can be involved in immune mediated adverse reactions.
authors.affiliation: Dipartimento di Scienze Farmacologiche e Biomolecolari (DiSFeB), Università degli Studi di Milano, Milan, Italy.
authors.forename: Daniele
authors.initials: D
authors.lastname: Corti
authors.affiliation: Dipartimento di Scienze Farmacologiche e Biomolecolari (DiSFeB), Università degli Studi di Milano, Milan, Italy. Electronic address: valentina.galbiati@unimi.it.
authors.forename: Valentina
authors.initials: V
authors.lastname: Galbiati
authors.affiliation: Dipartimento di Scienze Farmacologiche e Biomolecolari (DiSFeB), Università degli Studi di Milano, Milan, Italy.
authors.forename: Nicolò
authors.initials: N
authors.lastname: Gatti
authors.affiliation: Dipartimento di Scienze Farmacologiche e Biomolecolari (DiSFeB), Università degli Studi di Milano, Milan, Italy.
authors.forename: Marina
authors.initials: M
authors.lastname: Marinovich
authors.affiliation: Dipartimento di Scienze Farmacologiche e Biomolecolari (DiSFeB), Università degli Studi di Milano, Milan, Italy.
authors.forename: Corrado L
authors.initials: CL
authors.lastname: Galli
authors.affiliation: Dipartimento di Scienze Farmacologiche e Biomolecolari (DiSFeB), Università degli Studi di Milano, Milan, Italy.
authors.forename: Emanuela
authors.initials: E
authors.lastname: Corsini
mesh_terms: D000485:Allergens / Q000009:adverse effects*; D051940:B7-2 Antigen / Q000378:metabolism; D001681:Biological Assay; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D004342:Drug Hypersensitivity; D006801:Humans; D018799:Intercellular Adhesion Molecule-1 / Q000378:metabolism; D016209:Interleukin-8 / Q000235:genetics / Q000378:metabolism; D064546:Protein Kinase C beta / Q000378:metabolism; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism
chemical_list: D000485:Allergens; D051940:B7-2 Antigen; C497964:CD86 protein, human; D016209:Interleukin-8; D018799:Intercellular Adhesion Molecule-1; D064546:Protein Kinase C beta; D048051:p38 Mitogen-Activated Protein Kinases
keywords: Cytokines; Drug allergy; PKC; Surface markers; p38 MAPK