[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.

[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.

[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.

[Cell-free pseudomonas vaccine. I. Enzyme properties and virulence of strains of P. aeruginosa; protective activity of their water-soluble antigens]. / Beskletochnaia psevdomonas-vaktsina. Soobshchenie I. Fermentativnye svoistva i virulentnost' shtammov P. aeruginosa, protektivnaia aktivnost' ikh vodorastvorimykh antigenov.

The enzymatic (protease, catalase and gelatinase) activity and virulence of the typing strains obtained from the Hungarian National Collection of Medical Bacteria, as well as those of highly toxigenic PA-103 strain and 2 newly isolated P. aeruginosa strains were studied. No correlation between these properties of the strains was observed. Aqueous extracts prepared from some of the typing strains contained soluble specific and non-specific protective antigens. The partially purified, low toxic, water-soluble antigen(s) of typing strain No. 170015 (serogroup 07 according to Lányi or immunotype 2 according to Fisher) protected mice against infection with the homologous strain(s) belonging to immunotypes 3, 4 and 7, and to O-serogroups 8, 10 and 13. The immunization of mice with a single injection of partially purified protective antigen No. 170015 ensured pronounced protection of the animals against P. aeruginosa infection.

[Protective properties of antigenic preparations of Pseudomonas aeruginosa]. / Protektivnye svoistva antigennykh preparatov Pseudomonas aeruginosa.

The protective activity of various Ps. aeruginosa cell antigens was studied, with the aim of isolating highly active components for a complex immunologic preparation against Ps. aeruginosa infection. The preparations of the supernatant fluid obtained by ultracentrifugation of slime or aqueous extract of Ps. aeruginosa were found to have the highest protective activity. Such preparations had low toxicity. Antigens with a molecular weight of 10,000 to 30,000 daltons obtained by the membrane fractionation of the aqueous extracts of acetone-treated bacteria, had also a high protective activity and were non-toxic for rats.

[Immunological study of the cellular components of Pseudomonas aeruginosa. VI. The toxicity and protective properties of its mucus and its fractions]. / Immunologicheskoe izuchenie kletochnykh komponentov sinegnoinoi palochki. Soobshchenie VI. Toksichnost' i protectivnye svoistva slizi i ee fraktsii.

Capsule-like mucus was obtained from two newly isolated mucoid strains of P. aeruginosa and then treated with ethanol. The mucus was fractionated by the method of differential centrifugation (at 15, 000 g for 1 h, at 105, 000 or 170, 000 g for 3 h) and gel chromatography in columns packed with Sepharose 4B. The sediment fractions contained 30--80% of high molecular polysaccharide protein (peptide) mucus components which were toxic for mice and protected 25--77% of rats against experimental P. aeruginosa infection. The supernatant fluid fractions contained 60--80% of predominantly protein components with molecular mass between 20,000 and 60,000 daltons. These mucus components were slightly toxic for mice and rats and protected 80--100% of rats against P. aeruginosa infection.

[Immunological study of cellular components of Pseudomonas aeruginosa. IV. Fractionation of Pseudomonas aeruginosa mucus by diafiltration and biological properties of isolated fractions]. / Immunologicheskoe izuchenie kletochnykh komponentov sinegnoioni palochki. Soobshchenie IV. Fraktsionirovanie slizi Pseudomonas aeruginosa metodom diafil'tratsii i biologicheskie svoistva vydelennykh fraktsii.

In fractionation of Pseudomonas aeruginosa mucus (strains No. 8 and 1463) by means of diafiltration on the system of membranes Diaflo XM-300, XM-100A, PM-30, and PM-10 there was obtained a successive series of fractions differing by the molecular weight and chemical composition. According to the results of gel chromatography fractions with the mol wt of 100000 dalton and over apparently represented protein-polysaccharide components of mucus in the form of complexes; fractions with the mol wt of 30000 dalton and lower contained a considerable amount of free protein along with the protein-polysaccharide complex. The fractions obtained differed by biological properties: fractions with the mol wt of 100000 dalton and over were toxic for mice and possessed weak antigenic properties in the precipitation in agar test and immunoelectrophoresis; fractions with the mol wt lower than 30000 dalton expressed in the mentioned test distinct antigenic properties and proved to be practically nontoxic for mice. Thus the use of diafiltration method permitted to separate the antigenic, weakly toxic component of Ps. aeruginosa mucus from the toxic factor with weak antigenic properties.

[Immunologic study of the cellular components of bacillus pyocyaneus. III. Immunochemical analysis, toxicity and protective properties of water-soluble antigenic complexes]. / Immunologicheskoe izuchenie kletochnykh komponentov sinegnoinoi palochki. Soobshchenie III. Immunokhimicheskii analiz, toksichnost' i protektivnye svoistva vodorastvorimykh antigennykh kompleksov.

Aqueous extracts of two Ps. aeruginosa strains killed with acetone were subjected to fractionation by preparative ultracentrifugation and gel-chromatography. Toxic activity of the extract was found to be connected with the high-molecular, possibly glycoprotein components reacting with the corresponding antiserum in the immunoprecipitation test, and protecting 30--40% of rats against the generalized infection with Pseudomonas aeruginosa. This protective activity is apparently connected with the protein components (molecular weight--20000--60000 dalton), nontoxic for mice, not reacting with the corresponding antiserum in the immunoprecipitation test, and protecting 60 to 80% of rats against Ps. aeruginosa infecsion. Thus, as a result of preparative ultracentrifugation and gel-chromatography it was postible to divide the toxic and the nontoxic protective components of Ps. aeruginosa.

[A study of pancrealysates of Sh. flexneri 2a and Sh. sonnei fractionated by filtration through Sephadex G-200 and ultracentrifugation]. / Izuchenie pankrealizatov Sh. flexneri 2a i Sh. sonnei, fraktsionirovannykh fil'tratsiei cherez sefaeks G-200 i ul'tratsentrifugirovaniem

A high-molecular fraction of the O-antigen of Sh. flexneri and Sh. sonnei was isolated from the antigen complex by gel-filtration through Sephadex G-200 and ultracentrifugation. The high-molecular fraction obtained by ultracentrifugation at 105000 g contained no low-molecular components.

[Immunological study of the cellular components of Pseudomonas aeruginosa. Report 1]. / Immunologicheskoe izuchenie kletochnykh komponentov sinegnoinoi palochki. Soobshchenie 1

Water extracts obtained from acetone culture of 5 strains of Ps. aeruginosa isolated from wounds of patients with severe burns were investigated. These extracts contained exotoxin and capsular mucous cell component. By the data of gel-chromatography the extracts proved to be heterogeneous. Their toxic activity was associated chiefly with the high-molecular fraction. The extracts were active in experiments of immunodiffusion and immunoelectrophoresis with homologous and heterologous antisera. Up to 5 components with a different electrophoretic mobility were found in the extracts. There proved to be a correlation between the content in the extract of protein and its toxicity for mice. Differences between the strains of various O-serological groups were detected in experiments of gel-chromatography.

[Molecular weight heterogeneity of isolated antigenic complexes and their immunochemical activity]. / Molekuliarno-vesovaia geterogennost' izolirovannykh antigennykh kompleksov i ikh immunokhimicheskaia aktivnost'

Antigenic complexes extracted from S. typhi Ty24446 in tryptic proteolysis displayed heterogeneity by molecular weight. The O-antigenic activity of the preparation was associated with the components with a mol. weight of 6.7-10(4)--2-10(6), Vi-antigenic activity--with the components with a mol. weight of 6.7-10(4)--2-10(6) and the components with a mol. weight of 1.75-10(4)--6.7-10(6). Components with a mol. weight below 1.75-10(4) possessed no Vi-antigenic activity. Correlations between the high- and low-molecular components in the "tryptic" antigen depended on the regimen of proteolysis.