In vitro diagnosis of contact allergy to nickel: The value of the ELISpot assay

R. Spiewak1, H. Moed2, B. M. E. von Blomberg2, D. P. Bruynzeel2, R. J. Scheper2, S. Gibbs2, T. Rustemeyer2

1Jagiellonian University, Medical College, Krakow, Poland

2VU University Medical Centre, Amsterdam, the Netherlands

Source: Spiewak R, Moed H, von Blomberg BME, Bruynzeel DP, Scheper RJ, Gibbs S, Rustemeyer T. In vitro diagnosis of contact allergy to nickel: The value of the ELISpot assay. J Invest Dermatol 2005; 125 (3, Supplement: 35th ESDR Annual Meeting, 22-24 September 2005, Tübingen, Germany): A52.

 

Background: Diagnosis of contact allergy is based on clinical data and patch tests. Among in vitro tests, lymphocyte proliferation test (LPT) is most frequently used. A disadvantage of LPT is that it is based on radiochemicals, which restricts its use only to laboratories with radionuclide facilities.

Objective: To find a cytokine secretion assay giving results that correlate best with clinical diagnosis and with LPT.

Methods: PBMC from 14 patients with ACD to nickel and 14 non-allergic controls were tested for their reactivity to nickel. In all subjects, patch tests and LPT with nickel sulphate were done. A range of non-radioactive secretion assays was performed, including ELISpot assays for IL-2, IL-5, IL-13 and IFN-γ, and ELISA for IL-5 and IFN-γ. Beside standard culture conditions, cytokine secretion was also measured in cultures favouring the development of Tc1/Th1 or Tc2/Th2 lymphocytes ("skewing" through addition of IL-7 with respectively IL-12 or IL-4).

Results: The best correlation with clinical diagnosis (patch tests and history) was observed for IL-13 ELISpot with Tc2/Th2 skewing (r=0.654, P<0.001), followed by LPT (r=0.612, P<0.001), and IL-5 ELISpot with Tc2/Th2 skewing (r=0.551, P=0.002). The non-radioactive method that correlated best with LPT was IL-2 ELISpot (r=0.809, P<0.001), followed by IL-13 ELISpot (r=0.778, P<0.001), and IL-5 ELISA (r=0.669, P<0.001). Interestingly, IFN-γ ELISpot and IFN-γ ELISA correlated very poorly with both clinical diagnosis and LPT results (r<0.010 in each case).

Conclusions: Results of IL-13 ELISpot with Tc2/Th2 skewing correlate best with clinical diagnosis of contact allergy to nickel, whereas IL-2 ELISpot seems a good non-radioactive alternative for lymphocyte proliferation test.

Poster
Handout (PDF)

Links:

ELISpot.biz - training and implementation

ELISpot.biz - scanning and analysis of ELISpot plates

Celimun - biomedical research outsourcing in immunology

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

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Document created: 25 September 2005, last updated: 1 September 2006.