Peculiarities of production technology and properties of natural animal alpha-interferon preparations.

Principally new biotechnology of production of natural animal alpha-IFNs was created. Normative technical documentation on industrial production of these preparations was transmitted to the plants of Ukraine for their industrial obtaining. The peculiarity of proposed technology consists in usage of non-traditional raw-material--lymphoid organs and tissues (perhaps spleen), and also in introduction of coinduction into industrial cycle with the help of camizole (compound of phenilimidazothiazole family) which increased the output of final product by 3-8 times. Obtained preparations were tested in vitro and in vivo experiments. As it was shown in both cases bactericidal and absorbed activity of monocytes and neutrophils increased by 2-5 times, antibody genesis sharply increased after immunization of pigs and calves by colibacteriosis vaccine: titers were higher by 3-8 times (comparatively with control), especially two month after revaccination, but they were more stable too. Fractions of 12 and 18 kDa responsible for antibacterial and antitoxic activity respectively were found in non-purified preparations of alpha-IFNs.

[The use of gamma-interferon for the prevention and treatment of infectious diseases in piglets and calves]. / Vykorystannia hamma-interferonu dlia profilaktyky ta likuvannia infektsiinykh khvorob porosiat i teliat.

Administration of the homologous natural gamma-interferon for prophylaxis of infectious diseases showed that 2-3 injections of gamma-interferon prevented infectious morbidity of more than 90% of sucking pigs and new-born calves, sharply increased their preservation to 90% and above, increased their productivity by 1/3. In conditions of industrial growing of young animals therapy by this preparation of pigs and calves with colibacteriosis and parainfluenza in comparison with control significantly decreased mortality of morbid young animals, accelerated their recovery and promoted the weight gain in pigs and calves.

Treatment and prophylaxis of some infectious diseases of swine and cattle using of natural alpha-interferons.

On the basis of experimental studies which proved stimulating action on absorbing and bactericidal function of blood phagocytes and antibody genesis the methods of therapy and prophylaxis of wide spread infections in modern live-stock (transmissive gastroenteritis of pigs, parainfluenza of calves, colibacteriosis of swine and cattle) were created. It was established, that optimal one-time therapeutical dose of homologous alpha-IFN for intramuscular administration to new-born pigs was 2000-4000 IU per head, prophylactic dose was 1000-2000 IU. Therapeutical doses of alpha-IFN for calves did not exceed 4000-6000 IU, prophylactic--4000 IU. As a rule, it is necessary to introduce the preparation three times with 48 h intervals. With the account of these data the large-scale commissional trials of these preparations were carried out.

Properties of natural animal gamma-interferons and peculiarities of their action.

According to new biotechnology natural gamma-IFNs of swine and cattle as well as of laboratory animals (rabbits, rats, mice) were obtained. By trials in vitro and in vivo the preparations of swine and cattle gamma-IFNs have been tested. It was shown, that in vitro and in vivo functional activity of phagocytes (monocytes and neutrophils) sharply increased. There were increased phagocytic number, phagocytic index, bactericidal action, including functional reserve, the difference between spontaneous and stimulated by Staphylococcus antigen levels of bactericidity. Additional treatment by gamma-IFNs of animals after immunization by colibacteriosis vaccine increased antibody genesis 3-8 times in comparison with the control, but these titers were more stable during two months after revaccination. Besides antiviral activity, preparations of these gamma-IFNs had antibactericidal and antitoxic action of wide spectrum of specificity. The studied preparations are perspective means at usage in infectious pathology of animals, leukoses, immunodeficiencies.

Influence of Bacillus subtilis sp. lectin on functional activity of phagocytes.

Intramuscular and intraperitoneal introduction of Bacillus subtilis sp. lectin to mongrel white mice is responsible for the increasing of values of functional activity of peripheral blood phagocytes and peritoneal macrophages of these animals. These results confirmed similar data obtained in vitro. Bactericidal activity of monocytes and neutrophils significantly increased at minimal doses of lectin (0.1-0.5 microgram/animal). But this increase was lower at middle doses (0.5-5.0 micrograms/animal) and again increased at higher ones (5.0-25.0 micrograms/animal). Dose-dependent effect of lectin on macrophages was similar, but it was observed at lower concentrations. Administration of lectin did not give increase of functional reserve of studied cells, but was responsible for the increase of the phagocyte number and phagocytic index of monocytes and neutrophils at relatively middle doses (1.0-5.0 micrograms/animal). Phagocytic activity of macrophages did not increase. Thus, lectin of B. subtilis sp. is the agent non-toxic during studies in vivo, stimulating functional activity of phagocytes and macrophages.

[Natural interferons: the methods of their isolation, the properties and characteristics of the preparations and their use in animal infections]. / Pryrodni interferony: sposoby oderzhannia, vlastyvosti ta osoblyvosti diï preparativ, ïkh zastosuvannia pry infektsiiakh tvaryn.

Data of studies carried out in the field of interferonology during the last 20 years which are based on the proposed new biotechnologies for production of natural alpha- and gamma-interferons (IFNs) have been analyzed. "Know-how" of these technologies consists in the usage of non-traditional source (spleen instead of blood) for preparing the suspension of immunocytes--producers of preparations, in introduction of the stage of "coinduction" when producing alpha-IFNs, use of bacterial lectins of apathogenic bacilli as inducers of gamma-IFN genesis. It was shown, that in vitro and in vivo the both types of preparations several times increased both absorbing and bactericidal activity of phagocytes of swine and cattle peripheral blood, 3-5 times increased antibody genesis in animals after their immunization by colibacteriosis vaccine. It was proved that immunomodulating ability of alpha-IFNs is significantly (3-5 times) higher than that for gamma-IFNs. In non-purified preparations of IFNs the fractions have been discovered which had antibacterial and antitoxic action of wide spectrum of specificity. Both preparations were effective for prophylaxis and therapy of not only viral, but also bacterial infections in animals: transmissive gastroenteritis of swine, parainfluenza of calves, colibacteriosis of pigs and calves, etc., and promoted the increase of animals productivity by 20-35%. If under hemoblastosis alpha-IFN caused remission, they could be effectively treated using homologous gamma-IFN.

Lectin of Bacillus subtilis sp. as overinducer of gamma-interferonogenesis.

It has been demonstrated experimentally that lectin of Bacillus subtilis sp. in comparison with generally accepted Con A, PHA and lectin of "gold rain" grass--Laburnum anagyroides M e d i k in trials on white mice of CBA line gave in 4 hours of induction maximal titers of gamma-IFN in blood serum of animals--153.6 +/- 17.0 IU/ml. Practically identical titers had been obtained after induction by lectin "gold rain", some lower--after Con A and PHA. At swine gamma-IFN synthesis optimal density of cell suspension must contain 2.5 + 10(7) immunocytes in 1 ml, owing to which it is possible to obtain the titer equal 1 : 2150. Materials with using of bacterial lectins at various degree of purification had shown that maximal titers in blood serum of mongrel white mice were registered at administration to animals of non-purified lectin, 4 times lower--at using of half-purified and purified lectins. Data of these trials in vivo were confirmed by materials of gamma-IFN induction by immunocytes of swine, cattle and even man.