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