Contact allergy to nickel: Improved in vitro detection of allergen-specific cellular response by Th2/Tc2 skewing of leukocyte cultures

R. Spiewak1,2, H. Moed1, B. M. E. von Blomberg1, D. P. Bruynzeel1, R. J. Scheper1, S. Gibbs1, T. Rustemeyer1

1VU University Medical Centre, Amsterdam, the Netherlands

2Jagiellonian University Medical College, Krakow, Poland

Source: Spiewak R, Moed H, von Blomberg BME, Bruynzeel DP, Scheper RJ, Gibbs S, Rustemeyer T. Alergia kontaktowa na nikiel: Stymulacja fenotypu Th2/Tc2 poprawia wykrywalnosc swoistej reakcji komorkowej w hodowlach leukocytow. Alergia Astma Immunologia 2005; 10 (Suppl 1: 7th Central and Eastern European Conference "Allergy Asthma Clinical Immunology"): 164.

 

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:

ELISpot.biz - ELISpot training and implementation

ELISpot.biz - scanning and analysis of ELISpot plates

Celimun - biomedical research outsourcing in allergy and immunology

immunologia.BIZ - odczynniki i aparatura: ELISpot, ELISA, cytometria

Celimun - badania biomedyczne na zlecenie

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Document created: 15 June 2005, last updated: 1 September 2006.