Prednosti upotrebe biotehnološkog postupka produkcije interferona-gama iz prečišćene suspenzije humanih leukocita
ДокторандStanković, Bratislav P.
Чланови комисијеMarjanović, Goran
МетаподациПриказ свих података о дисертацији
Interferons (IFN) are first described in the late fifties of the twentieth century as special proteins in the family of cytokines and induced by the host, after invasion virus. If you are transferred to the fresh tissue was observed antiviral activity, thanks to inhibit replication of the virus. In this appearance immunology known as "interference". Under normal conditions in the human body concentration of IFN-a was below detectable levels or because cells synthesize IFN or contain as its constituent element. IFN was divided in three classes: IFN-, IFN- and IFN-. Most mammalian cells formed in unequal quantities of IFN- and IFN- after stimulation with viruses or other agents, or inducers of the synthesis of IFN. In many countries, is still detained biotechnological process for the production of human IFN (hlFN) for the clinical application of human leukocytes allocated from canned whole blood units, not older than 6 hours. After centrifugation the procedure centrifugation devotes... a buffy coat (leukocyte suspension) as a "side" -produkat. Further processing of "buffy coat" (BC) was purified from the red cell contamination, in order to obtain as pure substrate of human leukocytes, which is subjected to viral infection in order to stimulate the synthesis of hIFN-. Different viruses are used as inductors synthesis hIFN-, but in practice most often used "Sendai" virus, cultivated in horion-alantoic membrane of chicken embryos. Induction of synthesis hIFN- played on temperature of +37 ° C and lasts 18 hours. Refined suspension of human tal and practical. In fundamental respects got the time sequence data on the morphology, viability and induction capabilities of leucocytes, or their ability under the influence of the inductor synthesis hIFN- (or lyophilized virus hemagglutinin) higher yields of interferon, in the new proposed terms. In practical terms should consolidate that, when using a total amount of BC and from all transfusion centers in the Republic of Serbia as a raw material for the production of IFN-, in order to produce sufficient quantities IFN-, as biotherapeutic products, to cover not only needs Republic of Serbia, but to ensure the export of certain quantities of these products in countries with insufficient financial resources. Conclusions: 1. Learn yield and enhanced the antiviral activity of hIFN- in the compositions prepared from the suspension of leukocytes obtained from the pools "buffy coat"s which are added to dissolve dextran and 10% 6% hydroxy ethyl starch high molecular weight compared to the leukocyte suspension prepared in the conventional (current) method. 2. Better and more efficient biotechnological production process hIFN-, compared to the current procedure of obtaining hIFN- according to the modified "Cantell's" method. 3. The human leukocytes used for the production of hIFN- move more and have a greater ability to induction, while the same suspension of leukocytes is used for the production of hIFN- according to the modified "Cantell's" method has a stronger capacity of cell viability and induction ability of human leukocytes. 4. The lyophilized phytohemagglutinin - PHA-P, used as an inductor for the synthesis of hIFN- , although weaker than inductor "Sendai" virus, is a good inducer of the synthesis of which has been successfully induced human leukocytes in purified suspension.