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The lysate and recombinant antigens of various production included informula of ELISA-test-systems were analyzed. The ELISA-test-systems are used for detection of IgG to Herpes simplex virus type I and II. For testing the panel of serums PTH 201 (BBI Inc.) were used. The samples of this panel contain antibodies to Herpes simplex virus type I and II in mixed titers. The 69 serums of donors were used too (17 samples had IgG to Herpes simplex virus type I, 23 samples to Herpes simplex virus type II and 29 samples had no antibodies to Herpes simplex virus). The diagnostic capacity of mixture of recombinant antigens gG1 Herpes simplex virus type I and gG2 Herpes simplex virus type II (The research-and-production complex "DiaprofMed") was comparable with mixture of lysate antigen Herpes simplex virus type I and II (Membrane) EIE Antigen ("Virion Ltd."). In the test-systems for differentiation of IgG to Herpes simplex virus type I the recombinant antigen gG1 Herpes simplex virus type I proved to be comparable with commercial analogue Herpes simplex virus-1 gG1M ("Viral Therapeutics Inc."'). At the same time, capacity to detect IgG to Herpes simplex virus type II in recombinant protein gG2 Herpes simplex virus type II is significantly higher than in its analogue Herpes simplex virus-2 gG2c ("Viral Therapeutics Inc.").

Soluble fibrin and D-dimer are the most specific markers of blood coagulation cascade and the threat of thrombosis. Two immunoassay test systems were designed using the fibrin-specific and D-dimer-specific monoclonal antibodies. The clinical trials of the test systems were carried out in Ukraine. The high informative value of soluble fibrin quantification as a prognostic indicator of the threat of thrombosis associated with hip replacement and endoprosthetics of the abdominal aorta was shown. Independent D-dimer quantification is less informative. Simultaneous quantification of soluble fibrin and D-dimer before operation and at different time intervals after it is required for the prediction of postoperative thrombotic complications and monitoring the efficiency of antithrombotic therapy. Only in this case it is possible to get information about the state of the balance between blood coagulation and fibrinolytic systems, and determine the degree of the threat of thrombosis.

Technology for producing biologically active recombinant alpha2b-interferon is based on creating a bacterial producer strain containing the cDNA of human interferon alpha. The authors have obtained two producers of recombinant alpha2b-interferon, the synthesis of the target protein in them occurs in the inclusion bodies. The schemes of isolation and purification of biologically active recombinant alpha2b-interferon have been developed. The drug purity was approximately 97-98%. Biological activity in the culture of sensory cells in the cytopathic test was 4.2*10(8)ME/mg.

Here we report the obtaining of suspension, callus and hairy root culture initiated from carrot plants of Nantskaya and Perfektzya variety with the highest level of recombinant human interferon alpha-2b accumulation exhibited the highest level of plant protein extract antiviral activity (up to 12.8 x 10(3) IU/mg TSP). The antiviral activity of callus extracts was significantly lower comparing to the activity of plant extracts from parent organisms. However, the antiviral activity level of suspension culture extracts (up to 4.42 x 10(3) IU/mg TSP) and Ri-root ones (up to 4.42 x 10(3) IU/mg TSP) appeared to be comparable to analogical data of antiviral activity of transgenic carrot leaf extracts, this way the described cultures could be possibly used for comparatively speedy obtaining of recombinant therapeutic protein for curing and preventing of virus diseases.

Nanocrystalline ceria possesses a unique complex of physical and chemical properties making it highly bioactive material. In this review, modern data on the action of nanocrystalline ceria on cells, micro- and macroorganisms are analyzed. Special attention is paid to the analysis of the factors affecting protective properties of CeO2 with respect to the living systems.

It is shown that the recombinant protein GST-HSV2gG, containing the immunodominant regions of glycoprotein G of HSV-2 is accumulated in the form of inclusion bodies or in soluble form in the Escherichia coli BL21 (DE3) cells. The ratio between protein fractions varied depending on the physiological state of cells before biosynthesis. The kinetic parameters of bacterial populations were determined by mathematical modeling of growth curves based on the Verhulst logistic function. It was established that the induction of biosynthesis in the growth acceleration phase (at OD600 = 0.3) with 0.1 mM IPTG gives the maximum yield of soluble protein (26.75 mg/l or 17.6 mg/g biomass). The target protein was purified using the immobilized metal ion affinity and affinity chromatography technologies. Antigenic activity of the soluble form of recombinant protein GST-HSV2gG, was significantly (three times) higher than that of the protein purified from inclusion bodies (p < 0.05) and was comparable with the activity of the commercial analog (p > 0.05), that allows using this product in the immunosorbent test kits for diagnosis of IgG to HSV-2.

Antiviral activity of nanosize cerium dioxide sols (CeO2) in animal cell culture has been studied. The inhibiting effect of the mentioned sols upon reproduction of vesicular stomatitis test-virus was demonstrated for the first time in case of preliminary 24-hour contact with cells lines L929 and EPT. The effectiveness of protective action depends on initial precursor and the way of obtained water nanosize nanosols.

The investigation of cytotoxicity and antibacterial activity of the novel thrombin inhibitors containing retro-D-sequences -D-Arg-D-Phe--modified by D-arginine amino group by the residues of lauric acid or chromone-contained substituent, in comparison with known cationic preservative Nalpha-lauroyl-L-arginine ethyl ester (LAE) have been carried out. It has been shown that compound Laur-D-Arg-D-Phe-OMe has a similar cytotoxicity with LAE, and Chrom-D-Arg-D-Phe-OMe has almost twice higher toxicity than it fatty moiety contained analogues. Antibacterial activity of Laur-D-Arg-D-Phe-OMe against Staphylococcus aureus and Bacillus subtilis is close in action to LAE. It is assumed that ability of thrombin inhibitors to suppress the growth of some microorganisms can be explained by their ability to suppress activity of trypsin-like serine proteinases, which participate in the infection process of Staphylococcus aureus and influence on Bacillus subtilis sporulation. These findings open new prospects for exploring efficient antimicrobial agents among synthetic low-molecular trypsin-like serine proteinase inhibitors.

We have analyzed the recent data about Toll-like receptors (TLRs) that play the most important role in host immune defense. TLRs involved in induction and modulation of innate and acquired immunity as the integrators of their reactions. Genetic disorders of TLRs or their signaling pathways components were noticed with different pathologies. TLRs are the ideal molecular target for the therapy of many diseases including inflammatory, autoimmune, allergic and tumor diseases.

The uninvestigated interferon (IFN)-inducing capacity of glycoside derivatives of N-acetylmuramoyl-L-alanyl-D-isoglutamine (MDP) has been studied. Most MDP glycosides tested in vitro were more active than the reference preparation MDP. Under the in vivo conditions three studied preparations: MDP, a-heptyl-MDP and beta-(2-methyl-3-phenylchromonyl-7)-MDP stimulated production of considerable amount of circulating IFN that evidences for the promising character of their further investigation as preparations immunostimulators.

Two strains which produce recombinant proteins--analogs of membrane immunodominant epitopes with molecular weights 17 and 47 kDa of Treponema pallidum were constructed on the basis of E. coli recipient BL21 (DE3)-Gold with phage T7RNA-polymerase expression system. The method of mathematical modeling of growth curve was used to determine kinetic parameters which characterized physiological state of the culture producer in the process of fermentation. Results of immunoenzymatic analysis of blood serum samples from patients with syphilis and healthy donors are presented.

Imunophenotic characteristic of the mesenchymal stem and progenitor cells, their tissue origin and functional peculiarities as well as immunological aspects of its transplantation are presented in the article. Also the address migration and the site-specific differentiation of mesenchymal stem cells its age-related and pathological process induced changes are discussed. Prospects of clinical use of mesenchymal stem cells for the treatment of degenerative arthritis, spinal cord injuries, myocardial infarction and other disorders are shown.

An original kit containing 17 clones of hybridomas, the producers of monoclonal antibodies (MCAb) against the Fc-fragments of human IgG, has been obtained. The possibility of using horseradish peroxidase conjugates of obtained MCAb as a part of ELISA test systems (Diaproph-Med, Ukraine) was comparatively studied. The application of peroxidase conjugates of two monoclonal antibodies (156C10 and 153H11) as part of five diagnostic kits was shown to provide a high specificity and a high sensitivity.

The paper deals with the influence of the adenovirus (Ad) and Epstein-Barr virus (EBV) on functional activity of lymphocytes, in particular, the production of alpha- and gamma-interferons, tumor necrosis factor (TNF) in conditions of mono- or double infection of B- and T-phenotype (CEM) lymphoblastoid cells. It is shown, that Ad, EBV or both viruses induce high enough levels of interferon on both lines of cells and in control epithelial cells. The lymphoblastoid cells infected by viruses deep ability to synthesize alpha- and gamma-interferons under the influence of the corresponding inducers (Newcastle disease virus and hemagglutinine). Nevertheless, the levels of their formation are not high. Rather high parameters of activity of the tumor necrosis factor (TNF) were revealed during a day in the initial B95-8 cells and superinfected Ad after the effect of LPS of E. coli. Their activity in CEM cells also did not depend on the infection type.

The thesis about importance of specific immunoglobulin preparations in treatment of infectious diseases of viral and microbial etiology, uncontrollable infections, caused by representatives of herpes viruses family in particular, is developed in the paper. The types of immunoglobulin preparations, their advantages, technology peculiarities has been considered; the experience of application, including that of the authors have been analyzed. The necessity and expedience of intensive development of this branch of immunobiotechnology in Ukraine is proved, especially in connection with a high level of infectious morbidity in this country, concrete recommendations are given.

The work deals with the results of experimental evaluation of the influence of some new modified derivatives of muramyldipeptide (MDP) on the course of staphylococcal infection in mice. The preparations under study were found to produce rapid elimination of bacteria from kidneys and the increase of phagocytic activity of blood macrophages in animals. At the same time MDP and its derivatives stimulated natural killer cells whose activity was inhibited during infection. The dependence between the structure of these compounds and their protective action in staphylococcal infection, as well as the increase of the natural immunity characteristics of the body was followed.

Experimental data have been presented which prove antibacterial efficiency of interferon preparations and inducers under infection processes evoked by Staphylococcus aureus, Salmonella typhimurium, Pseudomonas aeruginosa, Klebsiella pneumoniae. It is shown that the system of phagocytizing cells and natural cells-killers plays the basic protective role in the organism under the persistence of these microbes. The basic regularities of activating effect of interferon preparations and its inducers on the functional activity of the above mentioned cells have been found. The methods of treatment with interferon drugs based on experimental data have been developed for the first time. They were used to cure patients with pyo-septic diseases.

Experiments on the CBA mice that had been within 30 km of the Chernobyl NPP accident for 3 months revealed a decrease in the phagocytic and bactericidal properties of peritoneal macrophages accompanied with a disturbance in the production of interleukin-1 and a tumor necrosis factor responsible for the cell immunity. A single administration of a silicon-containing enterosorbent Aerosil-350 expedient to remove radionuclides and ensure the organism detoxication made it possible to correct and restore the affected cell and organism immunity for 7 days.

Anti-HIV activity of the molecular complex forming during interaction between yeast RNA and tilorone was studied in vitro on the models of acute and chronic infection. Addition of this agent to the cells infected with HIV-1/IIIB and HIV-1/BRU decreased the virus reproduction controlled by assessing the viability of cells, syncytium production, and accumulation of p24 antigen in culture medium. The authors hypothesize that the detected anti-HIV effect is due to its capacity to produce type I interferon and direct antiviral action.

Cytokines (interferons, tumour necrosis factor and interleukin-1) with marked polyfunctional properties are described. They are synthesized by different cells of both immune and non-immune systems of organism and are able to stimulate, induce or inhibit various immunological and biological reactions and processes. The effect exerted by these cytokines on the course of bacterial infections is analyzed. The cytokines possess protective properties which are demonstrated in activation of neutrophils, intensification of bactericidal and phagocytic activity of macrophages and natural cells-killers.

The action of recombinant gamma-interferon (rINF-gamma) and tumor necrosis factor (rTNF-alpha) as well as that of their combinations was studied in a model of staphylococcal infection. It was observed that the use of rINF-gamma alone was not sufficient for the activation of the effector antibacterial function of macrophages with respect to staphylococci. At the same time it was shown that rTNF-alpha and rINF-gamma had a synergistic action on the process of the infection in mice evident from an increase in the pathogen elimination in the host. The use of the combination in the optimal doses increased the bactericidal activity of macrophages in the infected animals.

Impairments in the interferon status of children with chronic and recurring diseases of the respiratory organs were investigated and its correction was shown possible with the use of alpha 2-interferon (reaferon). Suppression of the interferon-producing capacity of the immunocytes was observed in all the cases of the disease aggravation. The level of the suppression correlated with the severity of the infection. In the patients affected by radiation the suppression of the interferonogenesis was more marked. However, in these cases no dependence of the detected impairments in the interferon status on the level of the radiation pollution of the territory of the children residence was shown. The inclusion of reaferon to the basal therapy resulted in a significant increase of the alpha-interferon production in the cases of bronchial asthma and asthmatic bronchitis as well as its recovery to the normal in the cases of recurring bronchitis and chronic pneumonia. In all the cases of bronchopulmonary diseases only a tendency towards normalization of the alpha-interferon indices was recorded.

Staphylococcus aureus cell-wall peptidoglycan has been shown to stimulate the bactericidal activity of peritoneal exudate macrophages. A more pronounced activating effect of peptidoglycan on the bactericidal activity of macrophages has been detected in the stimulated test. Peptidoglycan in doses of 5-100 mg induced interferon production by intact mouse splenocytes after 24-hour exposition with it. Peptidoglycan has been found to produce the most intensive effect on interferon production when injected intraperitoneally in a dose of 100 mg after 4-hour exposition.

In the complex of treatment of purulent-septic diseases leukinferon at a dose of 300 IU/kg was used according to the following scheme: 3 injections with the 48 h interval between injections. The effect of leukinferon mainly on the function of neutrophil granulocytes and subpopulation of T-lymphocytes was established. Together with clinico-laboratory data indicative of the effectiveness of treatment, normalization of interferon status and decrease in the level of a circulating factor of tumour necrosis in seriously ill patients was observed.

The influence of natural and recombinant mouse gamma-interferon (gamma-IF) on Salmonella typhimurium was experimentally studied. The preparations of gamma-IF were introduced into animals in a dose of 250 units per mouse according to different schedules before and after Salmonella inoculation. In these experiments the capacity of the animals for survival, the elimination of the infective agent from the body and the functional activity of peritoneal exudate macrophages were studied. Natural and recombinant gamma-IF was found to stimulate the ingestion activity and oxidizing metabolism of macrophages of the infected animals. At the same time the preparations of gamma-IF were not capable of enhancing the intracellular killing of salmonellae and tumor cells. Recombinant gamma-IF did not prevent the lethal outcome of S. typhimurium infection in mice and did not increase the elimination of the infective agent from the body. Natural gamma-IF, when introduced into mice in a dose of 250 units per mouse on days 1, 3 and 6 after Salmonella inoculation, ensured the survival rate of more than 80% of the animals, but did not essentially influence the dynamics of the elimination of the infective agent from the host body.

The effect of leukinferon on the system of phagocytes in infants with acute pneumonia was studied. There was observed an inhibition of the functional activity of neutrophils and monocytes in the blood of the infants. The inhibition level depended on the severity of the disease and the baseline immunological status of the infants. Normalization of the functional activity of the peripheral blood phagocytes led to more rapid discontinuation of the inflammatory process in the lungs. Leukinferon is a modulator of the phagocyte system and its action on the monocytes and neutrophil granulocytes was the higher, the lower was the initial functional activity of the phagocytes in the sick infants. The use of leukinferon in the complex therapy of the infants with acute pneumonia resulted in their more rapid recovery.

Porcine immune interferon (IF-gamma) was generated by stimulation with bacterial lectins. Physicochemical analysis showed porcine IF-gamma to be a glycoprotein with an isoelectric point around 8.4-8.8 and an apparent molecular weight of 28000 to 30000. A purification process used in this work included sequential steps of precipitation with ammonium sulphate and chromatographic separation on GPG-glass, con A-Sepharose and biogel P-100. This purification process gave a preparation of IF-gamma with specific activity of around 7 x 10(4) units per mg of protein.

The pathogenetic substantiation and technique of intrapleural sorption detoxication (IPSD) based on the use of high-dispersed silicone dioxide (trade mark "AEROSIL") are presented. Use of IPSD in the complex of treatment of 25 children with complicated forms of acute destructive pneumonia permitted to shorten the duration of intoxication syndrome, sanation of suppurative foci, stay of the patients at a hospital.

In staphylococcal infection the changes in functional ability of macrophages occur: their oxygen-depending bactericidity and adenosine-desaminase activity are depressed 5-nucleotidase ability increases. Introduction of homologous alpha-IFN in the dose of 1 x 10(3) u/mouse leads to enhancing macrophage bactericidity of the animals infected, inhibits their 5-nucleotidase activity and enhances adenosine desaminase one. Influence of alpha-IFN on the activity of adenosine metabolism enzymes in macrophages can be considered one of the most important mechanisms of its modulating effect in bacterial infections.

The results of the experimental study on the effect of the natural and recombinant gamma-interferons (gamma-IFs) of mice on the process of the infection caused by Klebsiella sp. are presented. The infection was reproduced by intraperitoneal contamination of mice with a virulent culture of Klebsiella pneumoniae 5055, line SHK. The gamma-IFs were administered to the animals in a dose of 250 units per mouse on days 1 and 3 after the contamination. Survival of the animals, clearance of the pathogen from the blood and liver and functional activity of the phagocytes in the contaminated mice were investigated. It was shown that both the natural and recombinant gamma-IF stimulated the phagocytic activity and oxidative metabolism of the phagocytes in the contaminated mice. Activation of these functions after the use of the natural gamma-IF correlated with its marked protective effect and accelerated elimination of the pathogen from the host which was not observed after the use of the recombinant gamma-IF.

The effect of type I interferons on the process of experimental salmonellosis in mice and rabbits, as well as their effect on salmonellosis in patients was studied. It was shown that homologous interferon increased the animal survival rate, activated bactericidal activity of the phagocytic cells and increased elimination of the infectious agents from the host. The complex of the etiotropic and pathogenetic treatment of 32 patients with salmonellosis included human leukocytic interferon, leukinferon. It was administered intramuscularly in a dose of 1 x 10(4)-2 x 10(4) IU: three injections at intervals of 48 to 72 hours (the treatment course). The course was repeated 10 days after the last injection of the first course. Addition of leukinferon to the routine scheme of the medicinal treatment of salmonellosis provided recovery of all the patients without complications or appearance of the bacteria carriers (the observation period of more than 3 years).

The study on mice with experimental generalized Klebsiella infection, carried out with the use of microbiologic, immunologic and pathomorphologic methods, revealed that the intraperitoneal injection of type I interferon into the animals prevented their death and led to the rapid elimination of the infective agent from their body, enhanced the phagocytic and metabolic activity of polymorphonuclear lymphocytes of their peritoneal exudate, decreased the manifestation of microcirculatory and dystrophic changes in the parenchyma of their internal organs.

The functional activity of phagocytic cells in 52 salmonellosis patients was studied with regard to the following characteristics: percent share of phagocytosis, phagocytic index, nitro blue tetrazolium test results, digestive activity. In patients with the unfavorable course of salmonellosis (the formation of carrier state) disturbances in the bactericidal activity of neutrophils and monocytes were established. For 32 patients leukinferon was included in the complex of etiotropic and pathogenetic treatment. The preparation was introduced in 3 intramuscular injections of 10,000 I. U. at intervals of 48 hours (the course of treatment); 10 days after the last injection this course was repeated. The use of leukinferon restored the normal functioning of phagocytes and the number of T-lymphocytes.

The effect of interferon, type 1, on the course of Salmonella infection in mice has been studied. The study has shown that the injection of homologous interferon of type 1 leads to the rapid elimination of the infective agent from the blood and organs of infected mice. Morphological study has shown that the injection of the preparations of interferon of type 1 may diminish pathological changes in the organs of the infected animals and the coagulation system of their blood.

The effect of type I interferon (IF) on the course of endotoxic shock was studied by morphological methods. It was found that preliminary (24 hours earlier) injection of preparations of homologous IF (10(3) ME) to ABLb/c and CC57 W mice sensitive to salmonella infection prevented the death of 90% of animals from a lethal toxic dose of the endotoxin or killed salmonellae. Preliminary injection of IF reduced sharply the intensity of morphological changes during all periods of the study. IF mitigates the toxic effect exerted on the cell and tissue structure both by increasing the functional activity of phagocytes, facilitating in this way elimination of the endotoxin from the body, and by increasing cell resistance to the direct effect of the bacterial toxin, which is just what determines its protective antitoxic effect. It was established that IF prevents the development of the most grave changes--thrombosis of the microcirculatory bed of vitally important organs by hindering the development of disseminated intravascular coagulation of blood, which determines the severity and outcome of shock.

The effect of the type I interferon on the development and process of experimental pyelonephritis caused by E. coli was studied on mice weighing 12 to 14 g. Interferon was administered intraperitoneally in a dose of 1000 units on days 3 and 7 of the disease. It was shown that the administration of the type I interferon to the mice with experimental pyelonephritis promoted rapid elimination of bacteria from the kidneys, prevented their penetration to the contralateral (intact) kidney, prevented marked macro- and microscopic damages in the kidneys, lowered the intensity of the inflammatory reaction, and increased the phagocytic activity of neutrophils and the number of the E-rosette-forming lymphocytes in the thymus. The data provided experimental grounding for clinical trials of interferon preparations in treatment of bacterial pyelonephritis.

Administration of interferon to mice with staphylococcal infection promoted elimination of the bacteria from their organs, influenced the immunological indices, normalized delayed type hypersensitivity, recovered the capacity of the spleen cells to produce interferon and increased the phagocytic activity of the peritoneal macrophages against the staphylococci. Addition of alpha-interferon to antibiotic therapy of patients with purulent septic infections of staphylococcal etiology markedly increased its efficiency. Improvement of the clinical picture of the disease, normalization of the general biochemical indices of the blood and elimination of the staphylococci from the blood and inflammation foci were observed.

A bacterial culture of E. carotovora 8638 lysed by virulent phage 62 after treatment with UV rays produces a moderate phage. The virulent and moderate viruses, antigenically unrealted, have a similar prolonged latent period (100 min) and the period of lysis (60 min). They also differ in the sizes of the capsids and the length of processes which are 750+/-30 and 2000+/-50A in the virulent phage and 600+/-30 and 1500+/-50A, in the moderate phage, respectively. The sedimentation constants are 602 and 340 S, respectively. In the virulent virus, the volume of the cavity of the protein capsid contianing 8 major and 5 minor peptides is twice as large as that in the moderate one (with 8 peptides); the molecular weight of their DNA shows approximately similar ratios: 52.53+/-17X10(6) and 29.79+/-0.88X10(6) daltons. Evidently, the moderate phage because of the reduced size of the genome contains less message for coding for structural proteins and nucleoprotein as a whole.

The antitoxic activity of leucocytic injection interferon I and immune interferon was shown in experimental erythrocyte hemolysis in the presence of staphylococcus alpha-toxin. The antitoxic effect was directly proportional to the interferon concentration in the medium and inversely proportional to the toxin concentration. Neutralization of the antiviral activity of leucocytic interferon did not lower its antitoxic effect. The highly purified and concentrated preparation of leucocytic injection interferon I and recombinant interferon had no antitoxic effect.

The influence of type I interferon on the persistence of S. typhimurium in the body of mice has been studied. The injection of the preparation of interferon has been shown to be conductive to the survival of the animals and to reduce the time of Salmonella persistence in the body. The injection of interferon enhances the phagocytic activity of macrophages in the peritoneal exudate of mice.

The preparations of alpha/beta- and gamma-interferons have been shown to stimulate the functional activity characteristics of mouse macrophages (phagocytosis, spreading, contacts with lymphocytes, bactericidal properties) obtained from the peritoneal exudate of intact animals and those infected with staphylococci. The immunomodulating action of gamma-interferon is more pronounced than that of the preparation of alpha/beta-interferon.