Diagnosis in vitro of contact allergy to nickel: Th2-skewing of PBMC cultures improves the detection of allergen-specific lymphocyte response

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, Dermatology & Pathology, Amsterdam, the Netherlands

Source: Spiewak R, Moed H, von Blomberg BME, Bruynzeel DP, Scheper RJ, Gibbs S, Rustemeyer T. Diagnosis in vitro of contact allergy to nickel: Th2-skewing of PBMC cultures improves the detection of allergen-specific lymphocyte response. Allergy Clin Immunol Int: J World Allergy Org 2005; 17 (Suppl 1: World Allergy Congress, June 26 - July 1, 2005, Munich, Germany): 20-21.

 

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 enzyme-linked immunospot assay (ELISPOT) , enzyme-linked immunosorbent assay (ELISA), and lymphocyte proliferation test (LPT).

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-γ 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-γ and IL-5, and LPT 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-γ 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. The best LPT performance was observed without addition of skewing cytokines.

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. It is possible that conditions in vitro favour the development of memory cells into Th2 phenotype. Moreover, Th2-skewing by addition of IL-7 and IL-4 further 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 might be due to the high spontaneous production of IFN-γ by various PBMC fractions (e.g. by NK cells), which possibly could obscure the Ni-specific IFN-γ secretion.

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