ELISpot in contact allergy: Th2-skewing improves detection of nickel-specific response in vitro

R. Spiewak*, H. Moed, B. M. E. von Blomberg, D. P. Bruynzeel, R. J. Scheper, S. Gibbs, T. Rustemeyer

Departments of Dermatology and Pathology, VU University Medical Centre, Amsterdam, the Netherlands

*Marie Curie Individual Fellow (European Community Fellowship QLK4-CT-2002-51504)

Source: Spiewak R, Moed H, von Blomberg BME, Bruynzeel DP, Scheper RJ, Gibbs S, Rustemeyer T. Elispot in contact allergy: Th2-skewing improves detection of nickel-specific response in vitro. 3rd EAACI Davos Meeting "Basic Immunology in Allergy and Clinical Immunology", 3-6 February 2005. SIAF 2005, 47 (Abstract 57).


The aim of this study was to determine whether skewing of lymphocytes from allergic patients towards Th1- or Th2-type could improve in vitro detection of nickel-specific cells using ELISPOT.

Methods: PBMC from 14 patients with allergic contact dermatitis (ACD) to nickel and 14 non-allergic donors (controls) were analysed for their response to nickel (Ni). IFN-gamma and IL-2 ELISPOT was performed without and with addition of Th1-skewing cytokines (IL-7 + IL-12). IL-5 and IL-13 ELISPOT was done without and with Th2-skewing cytokines (IL-7 + IL-4). Parallel to this, ELISA for IFN-gamma and IL-5 was carried out.

Results: Without skewing, the median number of cells secreting IL-5 in response to Ni was 10 cells per 1 million PBMC in ACD patients and 0 in non-allergic controls (p=0.007). After Th2-skewing, the numbers were 140 in ACD and 15 in controls (p=0.001). A similar pattern was found in IL-13 ELISPOT. In ELISA, median IL-5 concentrations were without skewing 14 pg/1 million PBMC in ACD and 0 in controls (p=0.001), and with skewing 266 and 33 respectively (p=0.014). The median number of cells secreting IL-2 in response to Ni was greater in ACD (67.5) than in controls (7.5; p=0.001), with no influence of Th1-skewing. As for Th1 response, there was a higher IFN-gamma output and more secreting cells among PBMC from ACD patients, but the difference to the controls remained non-significant both with and without Th1-skewing.

Discussion: It is generally accepted that ACD is associated with Th1-type response to allergens. A few recent studies showed that Th2 response could also be detected. Our results suggest that Th2 response in vitro discriminates Ni-allergic from non-allergic persons better than Th1 response. Moreover, Th2-skewing amplifies allergen-specific IL-5 response, expressed as an increase in both the number of secreting cells and the amount of secreted cytokine. The difficulty of detecting a significant Th1 response may be due to the high spontaneous production of IFN-gamma by PBMC, which possibly could obscure the Ni-specific secretion.


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Document created: 3 July 2005, last updated: 1 September 2006.