Generation and characterization of antigen-specific CD4+ , CD8+ , and CD4+ CD8+ T-cell clones from patients with carbamazepine hypersensitivity

• Published in: Journal of allergy and clinical immunology 2007-04, Vol.119 (4), p.973-981
• Main Authors: Wu, Ying, MSc, Farrell, John, BSc, Pirmohamed, Munir, MRCP, Park, B. Kevin, PhD, Naisbitt, Dean J., PhD
• Format: Article
• Language: English
• Subjects: Adult; Aged; Allergy and Immunology; anticonvulsant hypersensitivity syndrome; Antigen Presentation; Biological and medical sciences; Carbamazepine - immunology; Carbamazepine - pharmacology; CD4-Positive T-Lymphocytes - immunology; CD4-Positive T-Lymphocytes - metabolism; CD4-Positive T-Lymphocytes - pathology; CD8-Positive T-Lymphocytes - immunology; CD8-Positive T-Lymphocytes - metabolism; CD8-Positive T-Lymphocytes - pathology; Cell Proliferation - drug effects; Cells, Cultured; Clone Cells; Cloning; Cytotoxicity; Cytotoxicity, Immunologic - drug effects; Drug allergy; Drug Hypersensitivity - immunology; Drug Hypersensitivity - pathology; Epitopes, T-Lymphocyte - immunology; Epitopes, T-Lymphocyte - metabolism; Experiments; Female; Fundamental and applied biological sciences. Psychology; Fundamental immunology; Genotype & phenotype; Humans; Immune system; Immunopathology; Immunophenotyping; Lymphocyte Activation - drug effects; Lymphocytes; Male; Medical sciences; Mortality; Patients; T cell receptors; T cells
• ISSN: 0091-6749; 1097-6825
• DOI: 10.1016/j.jaci.2006.12.617

Background Hypersensitivity is a serious manifestation of anticonvulsant therapy characterized by infiltration of the epidermis and dermis by activated CD8+ and CD4+ T-cells, respectively. Attempts to characterize drug-specific CD8+ T cells have been largely unsuccessful. Objectives The aim of these studies was to generate and characterize CD4+ , CD8+ , and CD4+ CD8+ T cells in patients with carbamazepine hypersensitivity. Methods Carbamazepine-specific T-cell clones were generated from 5 patients by using modified cloning methodologies. Cell surface receptor phenotype, functionality, and mechanisms of antigen presentation were then compared. Results Ninety CD4+ , 23 CD8+ , and 14 CD4+ CD8+ carbamazepine-specific T-cell clones were generated. CD4+ T-cell clones proliferated vigorously with carbamazepine associated with MHC class II but exhibited little cytotoxic activity. In contrast, most CD8+ T cells proliferated weakly but effectively killed target cells via an MHC class I or MHC class II restricted, perforin-dependent pathway. CD4+ CD8+ T cells displayed characteristics similar to those of CD4+ T cells; however, drug stimulation was demonstrable in the absence of antigen-presenting cells. Carbamazepine was presented to CD4+ , CD8+ , and CD4+ CD8+ T cells in the absence of antigen processing. Drug stimulation resulted in the secretion of IFN-γ and IL-5. A panel of CD11a+ CD27− clones differentially expressed the receptors CXCR4, CCR4, CCR5, CCR8, CCR9, and CCR10. Conclusion Carbamazepine-specific CD4+ , CD8+ , and CD4+ CD8+ T cells exist in the peripheral circulation of hypersensitive patients, often many years after the resolution of clinical manifestations. Clinical implications Carbamazepine-specific CD4+ , CD8+ , and CD4+ CD8+ T cells displaying different effector functions and homing characteristics persist in hypersensitive patients' blood for many years after resolution of clinical symptoms.