Naslov
Immunohistochemical localisation of interferon in the cells

Autori
Filipič, Bratko ; Keše, Darja ; Rode, Bojan ; Lacković, Gordana

Izvornik
Periodicum biologorum. Supplement (0353-9164) 88 (1986); 301-302

Vrsta, podvrsta i kategorija rada
Radovi u časopisima, članak, znanstveni

Ključne riječi
immunohistochemical; interferon; cell

Sažetak
Interferons are glycoproteins which are synthesized in the cells after an induction or an infection. After the synthesis is finished, they are released from the cells, and in the extracellular space they bind to other cells. Concerning the pleiotropism in interferon activity, it seems that the most important step lies in its binding to the cell receptors on the plasrna membrane. As regards opinions and facts, does interferon penetrate into the cells differentby? Recent data has shown that some interferons can penetrate into the cell. Our experiments were designed to show, by different methods, (PAP - Peroxydase-antiperoxydase, IF - immunofluorescence) that human alpha interferon can be detected inside the cell. Throughout the experiments human embryonal fibroblasts were cultivated on small glass slides using Eagle's medium supplemented with 10% Foetal Calf serum (Flow). Two days before the cell treatment, Eagle's medium supplemented with 2% Foetal Calf serum was added to the fibroblast culture. In the experiments the cells were treated with human leucocyte interferon (having specific activity 104-105 units/mg) in the concentration of 1000 units/ml. The treatment was performed for 30, 60, 90 and 180 minutes (Mock in control). After the treatment the cells were fixed with 4% paraformaldehyde (for PAP) and with ice acetone (for IF). The cells were then washed twice with phosphate buffer saline (PBS) (pH 7.2) and the n treated with 0.05% Nonidet P-40 for 20 minutes. After additional washing in PBS, the cells were treated with anti-interferon serum (rabbit) diluted 1:500, and then washed again with PBS. The PAP reaction was performed using standard methods. To compare the PAP, the IF was performed as follows: to the treated cells sheep-antirabbit IgG conjugated with flouresceinisotiocianate (FITC) (1:50) (Inep-Zernun) for 45 minutes at 37 °C. Cells were then washed again with PBS and analysed using the fluorescence microscope (OPTON). Obtained results show that after the cell treatment with interferon the postive reaction (PAP) can be seen after 60 minutes around the cell nucleus ; thereafter, the positive reaction can also be seen in the nucleus. It can be concluded that after the cell treatment homologus interferon can penetrate into the cell. In connection with this, the question arose, did the complete molecule of interferon or only its different parts penetrate into the cell?

Izvorni jezik
Engleski

Znanstvena područja
Biologija

POVEZANOST RADA