Laboratory diagnosis of allergic contact dermatitis to nickel: The benefits of combining the highly sensitive ELISPOT assay with modified cell culture conditions

Radoslaw Spiewak, Heleen Moed, B. Mary von Blomberg, Derk P. Bruynzeel, Rik Scheper, Susan Gibbs, Thomas Rustemeyer

Celimun Biomedical Research, Krakow, Poland; Department of Dermatology, Free University, Amsterdam, The Netherlands

Source: Spiewak R, Moed H, von Blomberg BM, Bruynzeel D, Scheper R, Gibbs S, Rustemeyer T. Laboratory diagnosis of allergic contact dermatitis to nickel: The benefits of combining the highly sensitive ELISPOT assay with modified cell culture conditions. J Physiol Pharmacol 2007; 58 (Suppl 4: First International Conference "Medical Biology and Human Life"): 86.

Introduction: One of the obstacles in developing a reliable in vitro method for the diagnosis of allergic contact dermatitis (ACD) is low sensitivity of the detection assays. This may be overcome by enhancing allergen-specific response of lymphocytes and/or applying more sensitive assays. In the present study, we have combined both approaches.

Aims: To establish culture conditions that enhance nickel-specific secretion of cytokines by lymphocytes, as measured with ELISPOT.

Material and methods: Leucocytes of 14 patients with ACD to nickel and 14 controls were analysed for their response to nickel (Ni) in the presence of various combinations of IL-7 (anti-apoptotic cytokine), IL-4 (Th2/Tc2 enhancer) and IL-12 (Th1/Tc1 enhancer). Production of IFN-gamma, IL-2, IL-5, IL-13 was measured using the highly sensitive Enzyme-Linked Immunospot (ELISPOT) Assay.

Results: Without cytokines added, the median number of cells secreting IL-5 in response to Ni was 10 per million in ACD patients and 0 in controls (p=0.007). In the presence of IL-7 and IL-4, the numbers were respectively 140 and 15 (p=0.001). A similar pattern was found for IL-13. The median number of cells secreting IL-2 in response to Ni was 67.5 in ACD and 7.5 in controls (p=0.001), with no influence of added cytokines. As for IFN-gamma, differences between ACD patients and controls remained non-significant in all conditions tested.

Conclusions: Testing nickel-specific response of Th2/Tc2 lymphocytes cultured in the presence of IL-7 and IL-4 with the use of ELISPOT assay, offers a relevant progress in the in vitro detection of contact allergy to nickel.

Related articles:

  1. Spiewak R, Moed H, von Blomberg BME, Bruynzeel DP, Scheper RJ, Gibbs S, Rustemeyer T. Allergic contact dermatitis to nickel: Modified in vitro test protocols for better detection of allergen-specific response. Contact Dermatitis 2007, 56 (2): 63-69.
  2. Spiewak R. Patch testing for contact allergy and allergic contact dermatitis. Open Allergy J 2008, 1: 42-51.
  3. Spiewak R. Atopy and contact hypersensitivity: a reassessment of the relationship using objective measures. Ann Allergy Asthma Immunol 2005, 95 (1): 61-65.
  4. Spiewak R. Allergic contact dermatitis in childhood - a review and meta-analysis. Allergologie 2002, 25 (7): 374-381.
  5. Spiewak R, Pietowska J, Curzytek K. Nickel: a unique allergen - from molecular structure to European legislation. Expert Rev Clin Immunol 2007, 3 (6): 851-859.

Links:

Celimun Biomedical Research - outsourcing in allergy and immunology

International Photopatch Test Course, Krakow 2009

English-speaking dermatologist in Krakow (Cracow), Poland

Deutschsprechender Hautarzt in Krakow (Krakau), Polen

Institute of Dermatology, Krakow, Poland

ELISpot.biz - training, implementation, outsourcing

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Document created: 17 October 2008, last updated: 4 July 2010.