Belyĭ IuF

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Tartakovskiĭ Is

Prozorovskiĭ Sv

Vertiev IuV

Gamalei Institute of Epidemiology and Microbiology

Svetlana A. Ermolaeva

Gamalei Institute of Epidemiology and Microbiology

Spitsyn Sv

Drobyshevskaia Ei

Ezepchuk IuV

Petrukhin Vg

K Liuk

Karpova La

Cooperative Institutions

Gamalei Institute of Epidemiology and Microbiology

Ministry of Health of the Russian Federation

National Research Centre

Russian Academy of Sciences

V.I.Shumakov Federal Research Center of Transplantology and Artificial Organs

Immune Regulation (United Kingdom)

Fraunhofer Institute for Applied Optics and Precision Engineering

Academy of Medical Sciences

Fraunhofer Society

Czech Academy of Sciences, Institute of Experimental Medicine

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Detection of clostridia toxin markers in different types of the course of acute intestinal infections

Giulazian Nm Belaia Of Belyĭ IuF Pak Sg

The purpose of the investigation was to study the detection rates of markers and the level of C. diffcile A and B toxins and C. perfringens type A enterotoxin in patients with acute intestinal infections (AII). Two hundred and seventy-three patients with AII of varying etiology were followed up. According to the clinical syndrome, the patients were divided into 3 groups: (1) patients with the gastroenteritic (GE) type; (2) those with the gastroenterocolitic (GEC) type; (3) those with enterocolitic (EC) type. The circulation of markers of C. difficile A and B toxins and C. perfringens type A enterotoxin was studied, by employing the immunological test systems in the coagglutination test using the plates. The higher levels of antigens of all toxins were identified in the acute period of the disease in the GE and EC types than in the GEC type. There was a short increase in the levels of antigens of the test toxins in the GEC type and a gradual decrease in the GE and EC types. By discharge from hospital, the markers of toxins (more commonly of C. diffcile A) were preserved in 16.4% of the patients mainly in the GEC type.

Clostridia

Clostridium perfringens

Etiology

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[The typing of Legionella pneumophila strains by using monoclonal antibodies to the cytolysin].

PubMed (1991)

Spitsyn Sv Barkhatova Oi Drobyshevskaia Ei Belyĭ IuF Tartakovskiĭ Is

Studies on the typing of L. pneumophila strains of serogroup 1, isolated from patients and environmental objects, have been made with the use of monoclonal antibodies (McAb) to cytolysin. The results of the comparison of the specificity of our McAb with that of a commercial set McAb obtained from the USA make it possible to recommend preparations based on McAb to cytolysin for the detection of L. pneumophila pathogenic strains of serovar 1. The use of FITC- and peroxidase-labeled McAb to cytolysin permits the reduction of the time necessary for the diagnosis of Legionella infections and the detection of the antigen in a dose of 10 ng/ml.

Cytolysin

Legionella

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[Pathogenicity factors of bacteria with glycosylating activity].

PubMed (2011)

Tartakovskaia Di Araslanova Va Belyĭ IuF

A and B toxins of Clostridium difficile, a-toxin of C. novyi, lehal toxin of C. sordellii, and TpeL toxin of C. perfringens belong to the group of the so-called large Clostridium toxins. These toxins modify low-molecular weight guanosine triphosphate-binding proteins of the Rho/Ras family by their glycosylation that results in inactivation of major signal pathways in eukaryotic cells. Lgt glycosyltransferases, a new group of pathogenicity factors also capable of inactivating eukaryotic substrates via glycosylation, have recently been identified in Legionella. They are transported into cytoplasm of eukaryotic target cells by type 4 secretory system of Legionella. After translocation, the enzyme inhibits protein synthesis by attaching glucose residue to Ser53 of 1A elongation factor. The available data suggest an important role of bacterial glycosylating factors in the action of pathogens causing infectious diseases.

Clostridium perfringens

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[Conditions for Legionella pneumophila cultivation that affect cytotoxin production].

PubMed (1986)

Belyĭ IuF Tartakovskiĭ Is Neustroeva Vv Vertiev IuV Ezepchuk IuV

Experiments in CHO cell cultures have demonstrated the cytotoxic action of culture filtrates obtained after growing L. pneumophilia strain Philadelphia I, serogroup 1, in a liquid culture medium. This cytotoxic activity has been found to differ in its character from that described in earlier works. The dynamics of the accumulation of cytotoxin in the culture medium has been studied, and the work aimed at the determination of the limits of some cultivation parameters, within which the maximum toxin formation is observed, has been carried out.

Strain (injury)

Liquid culture

Liquid medium

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[Autoregulation of expression of secreted proteins in Listeria monocytogenes].

PubMed (2001)

Svetlana A. Ermolaeva Belyĭ IuF Tartakovskiĭ Is

The spectrum of proteins secreted by L. monocytogenes greatly depends on the composition of the cultivation medium. The introduction of activated charcoal (AC) into brain heart infusion (BHI) leads to the secretion of a number of additional proteins with mol.wt. ranging between 20 and 100 kD, whose production is not observed in pure BHI. The effect depends on the absorption capacity of AC: when adsorption capacity is reduced due to a decrease in the concentration of AC or its preliminary saturation with the components of the cultivation medium a drop in the level of the production of additional proteins is observed. The preliminary treatment of the medium with AC with its subsequent elimination prior to inoculation doses not change the spectrum of secreted proteins, though greatly inhibits the growth of L. monocytogenes. The data obtained in this investigation indicate that the effect produced by AC is based on the elimination of some product of L. monocytogenes vital activity from the cultivation medium; this product acts as the autoregulator of the synthesis of a number of secreted proteins.

Brain heart infusion

Secretory protein

Activated Charcoal

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[Monoclonal antibodies to Legionella pneumophila cytolysin].

PubMed (1989)

Spitsyn Sv Barkhatova Oi Al Drobyshevskaia Ei Belyĭ IuF Tartakovskiĭ Is

The fusion of spleen cells, taken from BALB/c mice immunized with the purified preparation of L. pneumophila cytolysin, with cells Sp2/0 and NP has been carried out. As a result, hybridoma cells producing IgG1, IgG3 and IgM antibodies to this protein have been obtained. All monoclonal antibodies (McAb) thus obtained react with L. pneumophila strain lysates in the precipitation test, while IgG3 and IgM antibodies react with erythrocyte diagnostic agents prepared from the lysate of L. pneumophila cells in the hemagglutination test. In the Western blot assay, McAb react with the 37 KD protein (cytolysin) and a number of other proteins from L. pneumophila cultures and L. pneumophila cell lysate, but do not react with the species-specific protein with a molecular weight of 29 KD, contained in the outer membrane of L. pneumophila, as well as with other species: L. bozemanii, L. dumoffii, L. longbeachae, L. micdadei. The possibility of using these McAb conjugated with FITC and peroxidase for the rapid diagnosis of Legionella infection is shown.

Cytolysin

Legionella

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[The purification and characteristics of listeriolysin O from Listeria monocytogenes].

PubMed (1994)

Karpova La Belyĭ IuF Tartakovskiĭ Is Prozorovskiĭ Sv

A scheme of the purification of listeriolysin O produced by L. monocytogenes strain NCTC 7973 was developed. The isolation procedure included the cultivation of the bacteria in heart-brain broth, the concentration of culture liquid free of bacteria with ammonium sulfate, cation exchange chromatography on a column packed with CM-Sepharose and Mono S, gel chromatography on a column packed with Superose 12. The preparation obtained with the use of this procedure was homogeneous, as confirmed by the data of SDS electrophoresis. The protein obtained in this investigation was no different from the protein studied earlier in its physico-chemical properties (molecular weight, heat stability, inhibition with thiole-active compositions, cholesterol, sensitivity to proteolytic enzymes) and corresponded to the characteristics of thiole-dependent hemolysins.

Listeriolysin O

Ammonium sulfate

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Citations (1)

[Cytolysin as an immunoserological marker of Legionella pneumophila].

PubMed (1988)

Tartakovskiĭ Is Belyĭ IuF Vertiev IuV Nagaev Ig Prozorovskiĭ Sv

Coagglutination test (CoaT), radioimmunoassay (RIA) and ELISA were used for detection of cytolysin produced by L. pneumophila. The sensitivity of RIA was 100 ng/ml, CoaT--10-20 ng/ml, ELISA--1 ng/ml. To determine cytolysin production among various strains and species of Legionella, the authors studied bacterial ultrasonic lysates. All 11 strains of L. pneumophila tested were cytolysin-positive. L. longbeachae, L. bozemanii and L. dumoffii strains failed to produce cytolysin. The authors believe that an antigen of cytolysin can be used for identification of L. pneumophila.

Cytolysin

Legionella

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[Production of recombinant fragment of VacA protein and development of non-invasive method for the diagnosis of Helicobacter pylori infection].

Belyĭ IuF Sheklakova La Vakhrameeva Ms Zhukhovitskiĭ Vg Petrukhin Vg

Vacuolizing toxin (VacA) Helicobacter pylori is an important factor of pathogenicity of Helicobacter pylori and a basic marker in the diagnosis of helicobacteriosis and related diseases. A coagulation-based diagnostic test-system was elaborated for the detection of VacA in clinical samples. A fragment of vacA was cloned, for the purpose, in Escherichia coli and expressed in preparative quantities; the coded protein was purified and used in raising the diagnostic serum. The thus designed coagulating test-system was successfully tested under the modeling conditions with clinical samples. Therefore, the designed express method can be used for the invasion-free determination of VacA in patients with gastric and duodenal pathologies.

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[The splitting of the acceptor proteins of the protein kinase system in eukaryotic cells by Legionella cytolysin].

PubMed (1991)

Belyĭ IuF Tartakovskiĭ Is Vertiev IuV Prozorovskiĭ Sv

In the process of protein kinase reaction carried out in the mixture consisting of tris-HCl buffer, EDTA, MgCl2, gamma-32P-ATP and the cytoplasmic fraction of rabbit pulmonary cells the phosphorylation of proteins with molecular weights of 150 and 55 kD took place. The addition of L. pneumophila culture fluid to the reaction mixture resulted in the splitting of phosphorylated proteins with the formation of the component having a molecular weight of 45 kD. These disturbances in protein kinase reaction were found to occur due to the involvement of Legionella cytoplasm, a previously characterized protein with a molecular weight of 37 kD, into the process. In this connection, the participation of cytolysin in the pathogenesis of Legionella infection may also be considered from the viewpoint of the effect produced by cytolysin on the regulatory processes affecting the metabolism of target cells.

Cytolysin

Legionella

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