[An acellular pertussis vaccine based on the natural complex of antigens isolated from the supernatant of a synthetic culture medium]. / Beskletochnaia kokliushnaia vaktsina na osnove prirodnogo kompleksa antigenov, vydelennykh iz supernatanta sinteticheskoi sredy kul'tivirovaniia.

A highly effective technology for obtaining a protective antigenic complex, including the main Bordetella pertussis protective antigens (pertussis toxin, filamentous hemagglutinin, agglutinogens, pertactin and adenylate cyclase) and isolated from the supernatant of B. pertussis cultivation medium, has been developed. A new method for the detoxication of antigenic complex which more available to preserve the protective property was suggested. Experimental batches of monovaccine and adsorbed DPT vaccine with the acellular pertussis component, possessing high protective activity and low histamine-sensitizing properties, have been obtained. The stability of protective properties both in liquid and lyophilized acellular pertussis preparations has been noted.

[The safety and protective properties of a new cell-free pertussis vaccine in animal trials]. / Issledovanie bezvrednosti i protektivnykh svoistv novoi beskletochnoi kokliushnoi vaktsiny v opytakh na zhivotnykh.

The preclinical trial of a new cell-free pertussis vaccine has been carried out with a view to its comparison with whole-cell pertussis vaccine in respect of the toxic and protective properties of the preparations under experimental conditions. The whole-cell pertussis vaccine is characterized by pronounced toxic action causing leukocytosis and histamine-sensitization, produces a negative effect on the detoxifying function of the liver and possesses sensitizing activity. The cell-free pertussis vaccine has pronounced protective activity. In animal experiments the supposed immunization dose of this vaccine has proved to be nontoxic.

[The use of monolaurates for removing endotoxins from preparations of cell-free pertussis vaccine]. / Ispol'zovanie monolauratov dlia udaleniia éndotoksinov iz preparatov beskletochnoi kokliushnoi vaktsiny.

The possibility of using monolaurates for removing endotoxin from acellular pertussis vaccines developed at the Laboratory of Immunomodulators, Mechnikov Research Institute for Vaccines and Sera (Moscow), has been studied. Monolaurates PEG-400 and PEG-600 obtained, respectively, from Fluka Chemie AG and Ferak GmbH (Germany) have been used. The use of monolaurate PEG-600 ensures the decrease of the toxicity of acellular pertussis vaccines by 2.34-6.3 times.

[The growth and protein A accumulation of Staphylococcus aureus A-676 on a medium made from nonfood raw material]. / Rost i nakoplenie proteina A Staphylococcus aureus A-676 na srede iz nepishchevogo syr'ia.

In this research the experimental analytical method of balancing culture media, based on the determination of the yield of the target product, i.e. staphylococcal protein A, has been used. The medium, developed in the course of this research work on the basis of a hydrolysate of protein-containing waste products of the tanning industry, makes it possible to achieve the growth of producer strains similar to that achieved in peptone-yeast medium. The parameters of the growth of S. aureus strain A-676 and the biological activity of protein A obtained in the newly developed medium have been studied.

[Cell-free Pseudomonas vaccine: its immunochemical characteristics and immunogenicity]. / Beskletochnaia vaktsina psevdomonas: immunokhimicheskaia kharakteristika i immunogennost'.

Two variants of cell-free protein vaccine have been prepared from the mixture of 4 P. aeruginosa strains, serovars 02, 06, 07 and 011, and from a single P. aeruginosa strain, serovar 02. The preparation contains proteins with molecular weight ranging from 20,000 to 100,000 and the admixture of lipopolysaccharide in negligible amounts (not exceeding 0.08% of dry weight). The vaccine produces no signs of toxicosis in laboratory animals. The vaccine effectively protects mice challenged with P. aeruginosa of different O-serotypes and stimulates the formation of specific protective antibodies in rabbits.

[Determination of ovalbumin-specific IgG antibodies in mouse sera by immunoenzyme analysis using horseradish peroxidase-conjugated staphylococcal protein A]. / Opredelenie spetsificheskikh IgG-antitel k oval'buminu v syvorotkakh myshei metodom immunofermentnogo analiza s ispol'zovaniem stafilokokkovogo proteina A, kon''iugirovannogo s peroksidazoi khrena.

The article deals with the possibility of using staphylococcal protein A conjugated with horse-radish peroxidase for the detection of specific IgG-antibodies to ovalbumin in mice by the indirect and competitive EIA techniques. Studies on specifying the parameters of the EIA system for the detection of specific IgG-antibodies are in progress.

[Immunoenzyme test system for the quantitative determination of tetanus antitoxin in vaccinated persons using horseradish peroxidase-conjugated staphylococcal protein A]. / Immunofermentnaia test-sistema dlia kolichestvennogo opredeleniia protivostolbniachnogo antitoksina u vaktsinirovannykh s ispol'zovaniem stafilokokkovogo proteina A, kon''iugirovannogo s peroksidazoi khrena.

An enzyme immunoassay (EIA) system for assays of tetanus antitoxin in vaccinees has been developed. As conjugate, staphylococcal protein A labeled with horse-radish peroxidase is used in this system. The possibility of using the newly developed EIA system in seroepidemiological surveys of the population is shown.

[Possibility of inactivation by formalin of staphylococcal protein A at different stages of its production]. / Izuchenie vozmozhnosti inaktivatsii formalinom materiala, soderzhashchego stafilokokkovyi protein A, na raznykh étapakh ego polucheniia.

The possibility of the formalin inactivation of material containing staphylococcal protein A in the process of obtaining the preparation has been studied. The inactivation of the material with formalin at a concentration of 5% (from the volume of the material) for 1 hour has been shown to ensure a complete bacteriostatic effect and the stability of the initial biological activity of staphylococcal protein A.

[Choice of the method for isolating conjugates of staphylococcal protein A with peroxidase for immunoenzyme analysis]. / Vybor metoda polucheniia kon"iugatov stafilokokkovogo belka A s peroksidazoi dlia immunofermentnogo analiza.

An attempt to obtain the preparations of peroxidase-labeled staphylococcal protein A, intended for use in the enzyme immunoassay, by the glutaraldehyde method has failed. The modified periodate method permitting the preparation of active conjugates of staphylococcal protein A with peroxidase has been developed.

[Determination of protein A I staphylococcus aureus strains isolated from monkeys]. / Opredelenei belka A u shtammov Staphylococcus aureus, vydelennykh ot obez'ian.

Protein A content in Staphylococcus aureus isolated from 6 species of monkeys at the Sukhumi Monkey Nursery has been studied. Protein A has been detected in 73% of the studied strains. One strain isolated from a rhesus macaque has been found to release high amounts of protein A into the environment.

[Effect of radiation sterilization on the immunobiological properties of the sorbed protective fraction of Bordetella pertussis]. / Izuchenie vliianiia radiatsionnoi sterilizatsii na immunobiologicheskie svoistva sorbirovannoi protektivnoi fraktsii kokliushnykh mikrobov.

The possibility of using radiation for the sterilization of the dried preparation of B. pertussis adsorbed protective fraction has been studied. The use of gamma irradiation in a dose of 1.5 X 10(3) J/kg has been found to permit obtaining sterile preparations while preserving their initial protective properties.

Immunochemical and immunological study of cell-wall proteins of Pseudomonas aeruginosa.

Crude aqueous extract was obtained from acetone-dried cells of Pseudomonas aeruginosa strain 868 (serogroup O2, Lányi and Bergan's schema) and subjected to ultracentrifugation (105,000 g, 3 h); the lipopolysaccharide (LPS)-containing precipitate was discarded and the supernatant containing water-soluble cell proteins was subjected to further fractionation. From a partially purified aqueous extract two fractions were obtained by step-wise precipitation with ammonium sulphate, namely, F1 (by 50% saturation), and F2 (by 80% saturation). By gel and ion-exchange chromatography from both fractions 9 subfractions were isolated differing in molecular weight, protein content, and LPS contamination. Subfractions 4 and 7 were practically free from LPS, and gave one precipitation line with antisera for strain 868. By immunoelectrophoresis subfraction 4 contained 2 cathodic and 1 anodic, whereas subfraction 7 mainly 1 anodic component. These subfractions were antigenically identical. With ELISA these subfractions were less active as compared to other subfractions, in particular to those of high molecular weight. The anti-subfraction 4 and anti-subfraction 7 sera were found to protect passively mice against intraperitoneal challenge by P. aeruginosa strain 8 (serogroup O2). These data support the authors' opinion that subfraction SF-4 and SF-7 are protective protein antigens (mol wt about 40,000 and 30,000, respectively), that are localized in the outer membrane of P. aeruginosa cell envelope.

[Optimal conditions for culturing Staphylococcus aureus strain A-676 to obtain biologically active protein A]. / Izuchenie optimal'nykh uslovii kul'tivirovaniia shtamma Staphylococcus aureus A-676 s tsel'iu polucheniia biologicheski aktivnogo proteina A.

Peptone-yeast and casein-salt culture media have been shown to be equally effective when used for the cultivation of S. aureus strain A-676 with a view to obtaining protein A. The seed dose has been determined and the optimum conditions for growing the producer strain have been established.

[Protective protein antigens of Pseudomonas aeruginosa]. / Belkovye protektivnye antigeny Pseudomonas aeruginosa.

Partially purified water extract was obtained from the initial water extract by ultracentrifugation. Nine protein fractions differing in molecular weight, homogeneity and the content of lipopolysaccharide (LPS) were obtained by stepwise precipitation with ammonium sulfate and subsequent fractionation in columns packed with Sephadex G-100 and DEAE cellulose. Two protein fractions with a molecular weight of 30000 and 40000 daltons were practically free of LPS. These fractions were homogeneous as shown by analytical centrifugation and formed a single precipitation line with P. aeruginosa antiserum; both fractions were found to be antigenically identical. In the enzyme immunoassay these two fractions proved to be least active in comparison with the other protein fractions, but when used for the immunization of rabbits, they induced the formation of specific protective (for mice) antibodies. Both antisera were equally active in the experiments of the passive protection of mice. The isolated LPS-free proteins are supposed to be the proteins of the outer membrane of P. aeruginosa cell wall and have the properties of protective antigens.

[Dynamics of staphylococcal infection in guinea pigs with delayed hypersensitivity to Staphylococcus aureus surface antigens]. / Dinamika stafilokokkovoi infektsii u morskikh svinok na fone povychennoi chuvstvetel'nosti zamedlennogo tipa k poverkhnostnym antigenam Staphylococcus aureus.

The relationship between delayed hypersensitivity (DH) to S. aureus surface antigens and the intensity of the infectious process induced by the sublethal infection of guinea pigs with S. aureus was studied. The protective effect, manifested by a decrease in the staphylococcal contamination of the spleen tissue and by an increase in the level of the activation of lymphocytes, was shown to correlate with DH induced by inactivated staphylococcal cells. In infected guinea pigs having DH to different staphylococcal antigens the disease either took a more severe course (in cases of DH to cell wall or peptidoglycan) than in the animals subjected only to infection, or no aggravation of the disease was observed (in cases of DH to protein A).

[Staphylococcal infection in guinea pigs with enhanced delayed hypersensitivity induced by Staphylococcus aureus surface antigens]. / Stafilokokkovaia infektsiia u morskikh svinok na fone povyshennoi chuvstvitel'nosti zamedlennogo tipa, indutsirovannoi poverkhnostnymi antigenami Staphylococcus areus.

A comparative study was made of the relationship between the DHS phenomenon and certain staphylococcal antigens and the intensity of infectious process in sublethal infection of guinea-pigs by Staphylococcus aureus. The protective effect, which was manifested by reduced amounts of staphylococcal colonies in spleen tissues and elevated level of lymphocytes, and neutrophil activation, correlated with DHS induced by surface antigens under study (cell wall, peptidoglycan, protein A). Incomplete combination or certain antigens induced DHS but did not increase the resistance to the following infection.

[Use of different media for growing the producer of the restriction enzyme XBA I]. / Ispol'zovanie razlichnykh sred dlia vyrashchivaniia produtsenta restriktiruiushchego fermenta XBA I.

The possibility of using culture media prepared from local ingredients and intended for growing the producers of restricting enzyme Xba 1 has been demonstrated. The yield of restricting enzyme Xba 1 per g of crude biomass, obtained with the use of peptone-yeast medium prepared from ingredients supplied by Difco Laboratories (USA), has proved to be 4 times greater than that obtained with the use of peptone-yeast medium prepared from local ingredients. At the same time the use of casein-saline medium ensures the yield of the enzyme, similar to that obtained with the use of peptone-yeast medium prepared from ingredients supplied by Difco Laboratories, but with a greater content of nonspecific nucleases.

[Elaboration of a method of stabilizing the protective fraction of pertussis microbes]. / Razrabotka metoda stabilizatsii protektivnoi fraktsii kokliushnykh mikrobov.

A method for stabilizing adsorbed preparations of the protective fraction of B. pertussis has been developed; according to this method, a colloid suspension of protective protein in phosphate buffer is obtained, the protein is then adsorbed on aluminium hydroxide gel and lyophilized with 10% of sucrose. If stored at 4 degrees C, these dried preparations have been found to retain their immunogenicity for 1 year.

[Cell-free Pseudomonas vaccine. IV. Laboratory trials of the effectiveness of experimental Pseudomonas vaccines]. / Beskletochnaia psevdomonas-vaktsina. Soobshchenie IV. Laboratornye ispytaniia éffektivnosti éksperimental'nykh psevdomonas-vaktsin.

Partially purified water-soluble cellular protein antigens have been obtained from 2 newly isolated P. aeruginosa strains and 1 museum P. aeruginosa strain, belonging to immunotypes 2, 3 and 7, by the method of preparative ultracentrifugation. Such trivalent P. aeruginosa vaccine (PV) has proved to be effective in direct and cross experiments of the active protection of mice. The method of ultrafiltration has been used to prepare monovalent PV from a newly isolated strain belonging to immunotype 3/7. This monovalent PV has been found to stimulate immunity to infection with homologous or heterologous P. aeruginosa strains in mice. The comparison of the results obtained in the study of PV prepared by the methods of ultracentrifugation and ultrafiltration suggests that both these methods for the isolation and purification of P. aeruginosa protective protein antigens are equally effective.