Detection of intracellular interferon-γ by light microscopy using an immunoperoxidase technique: correlation with the corresponding mRNA and protein product

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Abstract
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.

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