Allergic contact dermatitis to nickel: Modified in vitro test protocols for better detection of allergen-specific response

Radoslaw Spiewak, Heleen Moed, Brigitta Mary E. Von Blomberg, Derk P. Bruynzeel, Rik J. Scheper, Susan Gibbs, Thomas Rustemeyer

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To date, no in vitro test is suitable for routine diagnosis of contact allergy. The aim of our study was to establish improved in vitro test protocol for the detection of antigen-specific responses of lymphocytes from patients with allergic contact dermatitis to nickel (Ni-ACD). Blood leucocytes from 14 Ni-ACD patients and 14 controls were cultured in the presence of 'cytokine cocktails' skewing lymphocytes towards 'type 1' [interferon-γ (IFN-γ)-secreting] or 'type 2' [interleukin (IL)-5 and IL-13-secreting] phenotypes. The cocktails consisted of IL-7 and, respectively, either IL-12 or IL-4. Cell responses to nickel were measured with enzyme-linked immunospot assay (ELISpot), enzyme-linked immunosorbent assay (ELISA), and lymphocyte proliferation test (LPT). Significant differences between patients with Ni-ACD and controls were found for the 'type 2' cytokines IL-13 and IL-5, with further increase of allergen-specific responses occurring when cultures were supplemented with IL-7 and IL-4. No significant differences were found for IFN-γ. The best correlate to clinical diagnosis was LPT with 'type 2' skewing (r = 0.739, P < 0.001), followed by IL-13 ELISpot with 'type 2' skewing (r = 0.654, P < 0.001). The non-radioactive method that correlated best with LPT was IL-2 ELISpot (r = 0.809, P < 0.001). Overall, we conclude that combining ELISpot assay with proposed modifications of culture conditions improves detection of specific lymphocyte responses in contact allergy to nickel.

Original languageEnglish
Pages (from-to)63-69
Number of pages7
JournalContact dermatitis
Issue number2
Publication statusPublished - 1 Feb 2007


  • Allergic contact dermatitis
  • Cytokine cocktails
  • Enzyme-linked immunosorbent assay (ELISA)
  • Enzyme-linked immunospot assay (ELISpot)
  • Lymphocyte proliferation test (LPT)
  • Lymphocytes
  • Phenotype skewing

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