engleski

Immunocytochemical localisatlon of human alfa interferon (IFN) inside the human embriona fibroblasts

Until recently the references to the interferon´s (IFN) ability to penetrate the cell had been very scarce. Most of the data were only indirect. The authors have proved in the experiments presented here that the presence of human alpha interferon in the cell can be detected using the PAP (peroxidase-antipcroxidase) immunochemical method. The cells used in the experiment were human embryonic Fibroblasts cultivated on small glass slides in Eagle's medium supplemented with 100/0 fetal calf serum (FeS) (flow). The day before the IFN treatment the cells were cultivated in Eagle's medium with 2% FeS and then treated with 500 and 1000 units/ml of IFN having the specific activity of 104-105 units/mg of protein. This time scale was used: 15, 30, 60, 90 and 180 minutes. After the IFN treatment the cells were washed with PBS (phosphate buffered saline, pH 7.4) and treated with anti-alpha IFN (rabbit antihuman IgG, 1:1000). After being fixed in 4% paraformaldehyde in PBS containing 1% sucrose, the specimens were washed in PBS and treated with 0, 05% NP40 for 20 minutes. The PAP treatment was perforrned by the standard procedure. These results were obtained: immunoreactivity of cells can be recognised after 30-60 minutes, Simultaneously the typical morphological changes occur, i. e. the changes in the shape and size, including the swelling of the cells. After 30 minutes there is a positive reaction around the nucleus and after 60 minutes inside it. The results obtained allow of the conclusion that at least one part of IFN molecule can pass into the cell. The future cxperiments IFN will be fractioned to establish which part of the molecule is responsible for the penetration into the cell.

immunohistochemistry; human alpha interferon; human; embriona fibroblasts

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