594 Dynamic cytokine profiles combined with ELISPOT assay are useful in immunologically confirming the dapsone hypersensitivity syndrome

Abstract

Background: Dapsone hypersensitivity syndrome (DHS) is induced by drug-specific T cells. The specific dynamic cytokine profiles of DHS are unknown and the diagnosis of DHS in clinical practice remains a challenge. Objective: To investigate the DHS specific dynamic cytokine profiles, and explore specific enzyme-linked immunospot (ELISPOT) assay in the diagnosis of DHS. Methods: 14 types of cytokine levels (IFN-γ, Fas Ligand, Granzyme B, IL-4, IL-5, IL-6, IL-10, IL-13, IL-15, IL-17A, IL-21, IL-22, IL-23, and TNF-α) in DHS patients and dapsone tolerant patients carrying HLA-B*13:01 were analyzed using Luminex Bioplex assay. The performance of an IFN-γ/Granzyme B/IL-5 three-color fluoroSpot ELISPOT assay and Single-color ELISPOT assays based on traditional enzyme-tagged reagents in immunologically confirming DHS was evaluated in 16 DHS patients, four dapsone tolerant patients and 12 healthy donors. Results: Dynamic cytokine profiles showed that IFN-γ, IL-5, IL-13, Granzyme B and TNF-α were specifically up-regulated expression in DHS. In 16 patients with DHS, 9 (56.25%) patients were positive on IFN-γ ELISpot, 11 (68.75%) patients were positive on Granzyme B ELISpot and 11 (68.75%) patients were positive on IL-5 ELISpot, and when combination of IFN-γ, Granzyme B and IL-5 ELISPOT, the sensitivity increased to 87.5% (14/16) with the specificity of 100%. onclusions: An IFN-γ/Granzyme B/IL-5 ELISPOT assay with sensitivity of 87.5% was established, which could be treated as a promising tool in the diagnosis of DHS.