DETECTION OF INTRACELLULAR INTERFERON-GAMMA BY LIGHT-MICROSCOPY USINGAN IMMUNOPEROXIDASE TECHNIQUE - CORRELATION WITH THE CORRESPONDING MESSENGER-RNA AND PROTEIN PRODUCT
Rjem. Dolhain et al., DETECTION OF INTRACELLULAR INTERFERON-GAMMA BY LIGHT-MICROSCOPY USINGAN IMMUNOPEROXIDASE TECHNIQUE - CORRELATION WITH THE CORRESPONDING MESSENGER-RNA AND PROTEIN PRODUCT, Journal of leukocyte biology, 54(6), 1993, pp. 545-551
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.