Cytokine production by dengue virus antigen-responsive human T lymphocytes in vitro examined using a double immunocytochemical technique

摘要: A number of studies suggest that cytokines may contribute to the pathogenesis viral infections, including dengue. In this study, we developed a double immunocytochemical method and characterized cytokine-producing cells in peripheral blood mononuclear (PBMC) dengue virus-immune donors after vitro stimulation with specific antigens. We found immunostaining using immunoalkaline phosphatase (Vector blue) for [interferon-gamma (IFN-gamma), interleukin (IL) -2, -4, -1alpha, -1beta, -6, tumor necrosis factor beta (TNF-beta), TNF-alpha] immunoperoxidase [diaminobenzidine (DAB)] cell surface markers (CD3, CD4, CD8, CD20, CD68) provided best distinction double-positive from single-positive or -negative cells. The IFN-gamma, IL-2, IL-4, TNF-beta-positive increased 2 3 days No very few were detected PBMC non-immune stimulated antigens immune control antigen. analysis showed mainly CD4+ CD8+ T produced these cytokines. results obtained by immunocytochemistry consistent cytokine levels culture medium assayed enzyme-linked immunosorbent assay. conclusion, technique is suitable detection characterization PBMC. Furthermore, support hypothesis antigen-stimulated produce play role virus infection.