In vitro drug allergy detection system incorporating human liver microsomes in chlorazepate-induced skin rash: drug-specific proliferation associated with interleukin-5 secretion

Br J Dermatol. 2001 Feb;144(2):316-20. doi: 10.1046/j.1365-2133.2001.04021.x.

Abstract

Background: Chlorazepate is a benzodiazepine often used for pre-operative anxiolysis. The central metabolite responsible for the pharmacological and probably for the adverse effects of most benzodiazepines, including chlorazepate, is N-desmethyldiazepam. We report a woman who developed a generalized exanthem 1 day after receiving chlorazepate and four other drugs related to anaesthesia for surgery of the larynx. Patch tests pointed to chlorazepate as the culprit drug for the skin rash.

Objectives: The purpose of this study was to detect drug allergy to chlorazepate or a metabolite in vitro by means of the lymphocyte transformation test (LTT), and to determine the concentrations of the T-helper (Th) 2-type cytokine interleukin (IL)-5 and the Th1-type cytokine interferon (IFN) -gamma in the culture supernatants.

Methods: We performed an LTT with peripheral blood mononuclear cells from the patient and a control, employing human liver microsomes containing cytochrome P450 enzymes as a metabolizing system, in parallel cultures. IL-5 and IFN-gamma concentrations in the culture supernatants were assessed by enzyme-linked immunosorbent assay.

Results: In the LTT, no T-cell reactivity was observed to the parent compound chlorazepate, whereas coincubation of the drug with human liver microsomes yielded proliferative T-cell reactivity, which was associated with secretion of IL-5 but not of IFN-gamma.

Conclusions: We conclude that addition of a metabolizing system may be advantageous for in vitro detection of T-cell reactivity to drug metabolites in the LTT.

Publication types

  • Case Reports
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Clorazepate Dipotassium / adverse effects*
  • Cytokines / biosynthesis
  • Drug Eruptions / diagnosis*
  • Female
  • Humans
  • Immunoglobulin E / blood
  • In Vitro Techniques
  • Interleukin-5 / metabolism*
  • Lymphocyte Activation
  • Microsomes, Liver / immunology*
  • Skin Tests / methods

Substances

  • Cytokines
  • Interleukin-5
  • Immunoglobulin E
  • Clorazepate Dipotassium