Enumeration of IFN-γ-producing human lymphocytes by spot-ELISA: A method to detect lymphokine-producing lymphocytes at the single-cell level
作者： Jannemieke M.T. Versteegen , Ton Logtenberg , Rudy E. Ballieux
DOI: 10.1016/0022-1759(88)90055-5
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摘要: Abstract We present a method to detect and enumerate individual interferon (IFN)-producing human lymphocytes. The assay is based on the ELISA-plaque developed by Sedgwick Holt (J. Exp. Med. (1983) 157, 2178; J. Immunol. Methods (1986) 87, 37). Mitogen-stimulated T cells are seeded in anti-IFN-γ-coated wells. After 16 h incubation period, removed. Subsequently rabbit anti-IFN-γ-antiserum followed goat anti-rabbit antiserum conjugated alkaline phosphatase used IFN-γ spots. Application of spot-ELISA combination with conventional ELISA reveals amount produced per cell. highly sensitive, easy perform rapid assay. Provided specific antisera available, this suitable production other lymphokines at single-cell level. To our knowledge, first report single, well-defined cell product measurement spot-ELISA.
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