DETECTION OF INTRACELLULAR INTERFERON-GAMMA BY LIGHT-MICROSCOPY USINGAN IMMUNOPEROXIDASE TECHNIQUE - CORRELATION WITH THE CORRESPONDING MESSENGER-RNA AND PROTEIN PRODUCT

Identifying individual cytokine-producing cells may help to acquire in sight into immunological processes. This study was designed to adapt a technique for the detection of individual cytokine-producing cells fr om an immunofluorescence to an immunoperoxidase staining procedure. Th e production of interferon-gamma (IFN-gamma) by anti-CD3-activated clo ned 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. Th e intracellular staining for IFN-gamma was preceded or paralleled by I FN-gamma mRNA production and followed by accumulation of IFN-gamma in the supernatant. It is concluded that intracellular IFN-gamma can easi ly be detected using an immunoperoxidase procedure. This procedure is highly sensitive and allows quantification of the production of multip le cytokines by counting the percentage of positively staining cells.