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Cytokine production in nickel‐sensitized individuals analysed with enzyme‐linked immunospot assay: possible implication for diagnosis
Author(s) -
Jakobson E.,
Masjedi K.,
Ahlborg N.,
Lundeberg L.,
Karlberg AT.,
Scheynius A.
Publication year - 2002
Publication title -
british journal of dermatology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.304
H-Index - 179
eISSN - 1365-2133
pISSN - 0007-0963
DOI - 10.1046/j.1365-2133.2002.04850.x
Subject(s) - elispot , peripheral blood mononuclear cell , immunology , cytokine , allergy , medicine , interleukin 4 , interferon gamma , t cell , immune system , biology , in vitro , biochemistry
Summary Background Patients with suspected allergic contact dermatitis still have to undergo patch testing for a correct diagnosis. As this has several disadvantages there is a need for additional methods, preferentially those that can be performed in vitro . Objectives To investigate the possibility of diagnosing contact allergy to nickel (Ni 2+ ) using the enzyme‐linked immunospot (ELISpot) assay that allows the analysis of cytokines at a single‐cell level in ex vivo activated peripheral blood mononuclear cells (PBMC). Methods Eleven female patients and nine age‐ and sex‐matched healthy volunteers participated in the study. All patients had a history of nickel allergy and a positive patch test reaction to NiSO 4 , while the controls' test was negative. PBMC were cultured in the presence or absence of NiCl 2 . Cell proliferation was measured with [ 3 H]thymidine incorporation, and the number of cytokine‐producing cells analysed with the ELISpot assay. Results The proliferative response of PBMC to Ni 2+ , expressed as stimulation index, was significantly higher in the nickel‐allergic patients than in the control group. Using the ELISpot assay, we found that PBMC from nickel‐allergic individuals responded to Ni 2+ with significantly greater production of interleukin (IL)‐4, IL‐5, IL‐13 and interferon‐γ, but not IL‐12, compared with the healthy controls. The number of IL‐4‐ and IL‐5‐producing cells correlated with the number of IL‐13‐producing cells in the nickel‐allergic patients, but Ni 2+ ‐induced PBMC proliferation did not correlate with the number of cytokine‐producing cells for any of the cytokines tested. Conclusions Our results indicate that the ELISpot assay could be a tool in the discrimination between nickel‐allergic and non‐allergic individuals.