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Background: Diagnosis of contact allergy is based on clinical data and patch tests. Efficiency of in vitro tests available to date is not sufficient for routine use. Objective: To study the influence of modified culture conditions on the efficiency of in vitro assays in detecting antigen-specific cell responses in allergic contact dermatitis to nickel (Ni-ACD). Methods: PBMC from 14 Ni-ACD patients and 14 controls were cultured in the presence of cytokine cocktails skewing T cells towards Th1/Tc1 or Th2/Tc2 phenotypes. The cocktails consisted of either IL-7 with IL-12 or IL-7 with IL-4, respectively. Cell responses to nickel were measured with ELISpot (IL-2, IL-5, IL-13 and IFN-γ), ELISA (IL-5 and IFN-γ), and lymphocyte proliferation test (LPT). Results of all in vitro assays were compared to clinical diagnoses. Results: Significant differences between Ni-ACD patients and controls were found for Th2/Tc2 cytokines IL-5 and IL-13, with a further significant increase of the allergen-specific response occurring when cultures were skewed towards Th2/Tc2. No significant differences were found for IFN-γ. The highest correlation with the clinical diagnosis was observed for LPT with Th2/Tc2 skewing (r=0.739, P<0.001), followed by IL-13 ELISpot with Th2/Tc2 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). Conclusions: Skewing cell cultures towards Th2/Tc2 phenotype and measuring production of Th2/Tc2 cytokines IL-5 and IL-13 significantly improves detection of nickel-specific responses in vitro. IL-2 ELISpot seems best non-radioactive alternative for LPT. |
Links:Sensimun - biomedical research outsourcing in allergy and immunology Sensimun - badania biomedyczne na zlecenie Institute of Dermatology, Krakow, Poland |
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Document created: 15 June 2005, last updated: 25 November 2021.