title: Allergic contact dermatitis to nickel: modified in vitro test protocols for better detection of allergen-specific response.
abstract: 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-gamma (IFN-gamma)-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-gamma. 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.
authors.affiliation: Institute of Dermatology, ul. Konarskiego 15, 30-049 Krakow, Poland.
authors.forename: Radoslaw
authors.initials: R
authors.lastname: Spiewak
authors.forename: Heleen
authors.initials: H
authors.lastname: Moed
authors.forename: Brigitta Mary E
authors.initials: BM
authors.lastname: von Blomberg
authors.forename: Derk P
authors.initials: DP
authors.lastname: Bruynzeel
authors.forename: Rik J
authors.initials: RJ
authors.lastname: Scheper
authors.forename: Susan
authors.initials: S
authors.lastname: Gibbs
authors.forename: Thomas
authors.initials: T
authors.lastname: Rustemeyer
mesh_terms: D000485:Allergens / Q000276:immunology*; D016022:Case-Control Studies; D046508:Culture Techniques; D017449:Dermatitis, Allergic Contact / Q000175:diagnosis* / Q000276:immunology* / Q000473:pathology; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006801:Humans; D007371:Interferon-gamma / Q000096:biosynthesis; D007378:Interleukins / Q000096:biosynthesis; D009532:Nickel / Q000276:immunology*; D011237:Predictive Value of Tests; D018417:Th1 Cells / Q000276:immunology*; D018418:Th2 Cells / Q000276:immunology*
chemical_list: D000485:Allergens; D007378:Interleukins; D009532:Nickel; D007371:Interferon-gamma