TY - JOUR
T1 - Allergic contact dermatitis to nickel
T2 - Modified in vitro test protocols for better detection of allergen-specific response
AU - Spiewak, Radoslaw
AU - Moed, Heleen
AU - Von Blomberg, Brigitta Mary E.
AU - Bruynzeel, Derk P.
AU - Scheper, Rik J.
AU - Gibbs, Susan
AU - Rustemeyer, Thomas
PY - 2007/2/1
Y1 - 2007/2/1
N2 - To date, no in vitro test is suitable for routine diagnosis of contact allergy. The aim of our study was to establish improved in vitro test protocol for the detection of antigen-specific responses of lymphocytes from patients with allergic contact dermatitis to nickel (Ni-ACD). Blood leucocytes from 14 Ni-ACD patients and 14 controls were cultured in the presence of 'cytokine cocktails' skewing lymphocytes towards 'type 1' [interferon-γ (IFN-γ)-secreting] or 'type 2' [interleukin (IL)-5 and IL-13-secreting] phenotypes. The cocktails consisted of IL-7 and, respectively, either IL-12 or IL-4. Cell responses to nickel were measured with enzyme-linked immunospot assay (ELISpot), enzyme-linked immunosorbent assay (ELISA), and lymphocyte proliferation test (LPT). Significant differences between patients with Ni-ACD and controls were found for the 'type 2' cytokines IL-13 and IL-5, with further increase of allergen-specific responses occurring when cultures were supplemented with IL-7 and IL-4. No significant differences were found for IFN-γ. The best correlate to clinical diagnosis was LPT with 'type 2' skewing (r = 0.739, P < 0.001), followed by IL-13 ELISpot with 'type 2' skewing (r = 0.654, P < 0.001). The non-radioactive method that correlated best with LPT was IL-2 ELISpot (r = 0.809, P < 0.001). Overall, we conclude that combining ELISpot assay with proposed modifications of culture conditions improves detection of specific lymphocyte responses in contact allergy to nickel.
AB - To date, no in vitro test is suitable for routine diagnosis of contact allergy. The aim of our study was to establish improved in vitro test protocol for the detection of antigen-specific responses of lymphocytes from patients with allergic contact dermatitis to nickel (Ni-ACD). Blood leucocytes from 14 Ni-ACD patients and 14 controls were cultured in the presence of 'cytokine cocktails' skewing lymphocytes towards 'type 1' [interferon-γ (IFN-γ)-secreting] or 'type 2' [interleukin (IL)-5 and IL-13-secreting] phenotypes. The cocktails consisted of IL-7 and, respectively, either IL-12 or IL-4. Cell responses to nickel were measured with enzyme-linked immunospot assay (ELISpot), enzyme-linked immunosorbent assay (ELISA), and lymphocyte proliferation test (LPT). Significant differences between patients with Ni-ACD and controls were found for the 'type 2' cytokines IL-13 and IL-5, with further increase of allergen-specific responses occurring when cultures were supplemented with IL-7 and IL-4. No significant differences were found for IFN-γ. The best correlate to clinical diagnosis was LPT with 'type 2' skewing (r = 0.739, P < 0.001), followed by IL-13 ELISpot with 'type 2' skewing (r = 0.654, P < 0.001). The non-radioactive method that correlated best with LPT was IL-2 ELISpot (r = 0.809, P < 0.001). Overall, we conclude that combining ELISpot assay with proposed modifications of culture conditions improves detection of specific lymphocyte responses in contact allergy to nickel.
KW - Allergic contact dermatitis
KW - Cytokine cocktails
KW - Enzyme-linked immunosorbent assay (ELISA)
KW - Enzyme-linked immunospot assay (ELISpot)
KW - Lymphocyte proliferation test (LPT)
KW - Lymphocytes
KW - Phenotype skewing
UR - http://www.scopus.com/inward/record.url?scp=33846459384&partnerID=8YFLogxK
U2 - https://doi.org/10.1111/j.1600-0536.2007.01045.x
DO - https://doi.org/10.1111/j.1600-0536.2007.01045.x
M3 - Article
C2 - 17244072
SN - 0105-1873
VL - 56
SP - 63
EP - 69
JO - Contact dermatitis
JF - Contact dermatitis
IS - 2
ER -