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Detection of intracellular interferon‐γ by light microscopy using an immunoperoxidase technique: correlation with the corresponding mRNA and protein product
Author(s) -
Dolhain Radboud J.E.M.,
Natalja Ulf AnderssoiV,
Haar ter,
Brinkman Brigitta M.N.,
Verweij Cornells L.,
Daha Mohamed R.,
Breedveld Ferdinand C.,
Miltenburg Andre M.M.
Publication year - 1993
Publication title -
journal of leukocyte biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.819
H-Index - 191
eISSN - 1938-3673
pISSN - 0741-5400
DOI - 10.1002/jlb.54.6.545
Subject(s) - immunoperoxidase , biology , immunofluorescence , staining , intracellular , cytokine , interferon , microbiology and biotechnology , messenger rna , monoclonal antibody , antibody , immunology , biochemistry , gene , genetics
Identifying individual cytokine‐producing cells may help to acquire insight into immunological processes. This study was designed to adapt a technique for the detection of individual cytokine‐producing cells from an immunofluorescence to an immunoperoxidase staining procedure. The production of interferon‐γ (IFN‐γ) by anti‐CD3‐activated cloned human T cells was used as a model system. After the conditions for the staining procedure were optimized, the immunoperoxidase technique was slightly more sensitive than the immunofluorescence technique. The intracellular staining for IFN‐γ was preceded or paralleled by IFN‐γ mRNA production and followed by accumulation of IFN‐γ in the supernatant. It is concluded that intracellular IFN‐γ can easily be detected using an immunoperoxidase procedure. This procedure is highly sensitive and allows quantification of the production of multiple cytokines by counting the percentage of positively staining cells.