The department’s director is N.A Zigangirova, Doctor of Biological Science.
The department was founded in 1979 on the basis of a number of laboratories with the common goal of research of the process of persistence of infectious disease agents and mechanisms, defining its role in the origin of acute, latent, chronic and slow forms of infection. Research is conducted into pathogens of varying types: bacteria, chlamydia, mycoplasma and viruses.
The defining characteristic of the last few years of the department’s work has been development of the fundamental issue of the pathomechanism on one hand and the wider use of research results in clinical practice connected to the enhancement of diagnosis, prevention and treatment of various infectious illnesses on the other.
Mycoplasma and L-form Bacteria Laboratory
The head of the laboratory is I.V. Rakovskaya, Doctor of Biological Science.
The main line of research is dedicated to the study of mechanisms of pathogenicity and long-term persistence of mycoplasma in the infected organism. Experimental models of urogenital and respiratory mollicutes, rheumatoid arthritis of mycoplasmal etiology and combined mycoplasmal and viral infections have been created for this. Several immunological mechanisms of mycoplasm “eluding” the immune surveillance of the host have been studied. At present, genetic and molecular mechanisms for persistence in mycoplasma are being studied.
It has been shown that in chronic respiratory mollicutes the expression of a range of genes is suppressed, including the gene coding adhesin PI, which is the main factor for the pathogenicity of mycoplasma. The mechanism and behavior of this process has been studied. A method has been developed that enables diagnosis of persistent mycoplasma during chronic infection that cannot be detected using standard methods, including PCR with the conventional method of DNA extraction. Research is being carried out into the relative pathogenicity of varying serovars of М.urealyticum.
The Center of the Ministry of Health and Social Development of the Russian Federation for mycoplasmosis functions as part of the laboratory, carrying out the following practical tasks:
analysis of disease incidence in the population;
development and enhancement of methods of diagnosis;
development of guidance documentation for diagnosis;
provision of consulting and methodological aid to staff of the State Sanitary and Epidemiological Service as well as to a number of organizations of the Ministry of Health and Social Development of the Russian Federation.
During completion of these laboratory tasks a set of diagnostic methods has been developed and comparatively evaluated, and a range of instruments for diagnosis of mycoplasma using immunofluorescence have been created:
In addition, diagnostic agents for detecting antibodies for М.hominis and М.pneumoaiae using enzyme immunodetection as well as for the detection of urogenital mycoplasma using PCR have been created.
Using the indicated diagnostic agents and a range of diagnostic methods, more than 7,000-8,000 patients with inflammatory diseases of the urogenital tract and respiratory system are investigated in the laboratory annually. The laboratory is constantly engaged in examining outbreaks of pneumonia among various patient populations across different regions of Russia.
The laboratory has a mycoplasma bank and a selection of hyperimmune serums that are constantly being expanded. The laboratory organizes annual seminars for healthcare workers. The laboratory provides constant training for lab doctors and scientific staff from numerous institutions around the country. Methodological recommendations are produced for diagnosis of mycoplasmal infections as well as practical guidance for doctors.
The head of the laboratory is Professor I.S. Tartakovskiy, Doctor of Biological Science.
The main lines of research are the study of regulatory mechanisms controlling the expression of pathogenicity factor at varying stages of interaction of the endocellular bacterial parasite with the host cell. The main models used in the laboratory’s research are the facultative endocellular parasites Legionella pneumophila and Listeria monocytogenes.
The following were isolated and characterized for the first time in the laboratory using the model of legionella: cytolysin (the main secretory protein of legionella with expressed cytotoxic and hemolytic activity) and UDP-glucosyltransferase, the activity of which correlates with the clinical significance of the detected strains. It has been established that the identified enzymes specifically deactivate components of the signal-carrying paths of eukaryotic cells. UDP-glucosyltransferase modifies 40-45 kD actin-like proteins of the cytoskeleton of macrophage-like cells via covalent attachment to the glucose target residue. Cytolysin reacts to the protein kinase activity of eukaryotes by way of specific partition of the phosphorylate eukaryotic proteins.
It was established for the first time based on the listeria model that the expression of the pathogenicity of factors of L. monocytogenes is under negative control, carried out by low molecular weight autocrine signaling produced by listeria and accumulating in the culture medium during the growth process. A collection of mutant strains of listeria with defects in the system of negative autoregulation have been obtained, with the constitutive hyperproduction of pathogenicity factors resulting in point amino acid substitutions in the sequence of the regulatory protein PrfA with impairments of certain functions of intracellular reproduction also obtained. The phenomenon of endocellular accumulation of synthesized factors of pathogenicity with the following mass of their secretion was also discovered. Methodological approaches have been developed that enable assessment of the level of expression for separate factors of the pathogenicity of listeria.
In addition to fundamental research, the laboratory also carries out applied research. Therefore, in connection with introduction by the Ministry of Health of the Russian Federation of hygiene standards that strictly regulate the content of pathogenic listeria in food, the laboratory is developing fast and inexpensive methods for the identification and typing of pathogens for the country’s sanitary and epidemiological services. It has been shown that PCR with primers directing the amplification of the p1cA gene fragment and the original bacteriological method based on the comparison of lecithinase activity of the culture in the presence of activated charcoal significantly increases the reliability and efficiency of rapid identification of L. monocytogenes.
By order of the Moscow City Government for reducing the risk of contact with pathogens and potential infection of the population with legionella, policies for microbiological monitoring, sanitary and hygiene assessment and the disinfection for technical means of centralized cleaning of air conditioning of premises in Moscow have been developed, in order to reduce the risk of contact with pathogens and potential infection of the population with legionella.
The laboratory is part of the Center of the Ministry of Health and Social Development of the Russian Federation for Legionnaires' disease and is in active cooperation with the WHO as part of the European Legionnaires' disease work group under the guidance of the EEC, uniting laboratories from 25 countries and carrying out epidemiological monitoring of Legionnaires' disease in Europe.
Regarding the issue of listeriosis, the laboratory is participating in an international EEC project (INTAS-2000-0471) entitled “Fundamentals of the rational control of food-borne listeriosis: a pan-European initiative”.
The leading researcher is Y.K. Kulakov, Doctor of Medical Science.
The main lines of research are the epidemiology of brucellosis, study of the infectious agent, and development and refinement of laboratory methods for diagnosis, prevention and treatment of brucellosis.
The laboratory carries out systematic analysis of the disease incidence of patients with brucellosis, monitoring of the epidemiological situation in the Russian Federation and its separate territories on the basis of existing record systems and its own data obtained from the laboratory. Monitoring of the temporal and spatial dynamics of the etiological structure of brucellosis is carried out with the aim of predicting an epidemic situation and giving early correction of the control system.
Regarding the issue of improving brucellosis diagnosis, the majority of attention is given to the study of the effectiveness of using the PCR method in the diagnosis of various forms of brucellosis in humans and in epidemiological practice. Research is carried out on a number of populations - those with acute and chronic forms of brucellosis, laboratory workers in contact with brucellosis cultures and those working for companies processing animal products. As a result, research has covered the correlation dependence of PCR and serological tests.
In order to improve the information value of criteria in the identification and differentiation of brucella-type representatives, the laboratory uses comparative analysis of protein profiles in polyacrylamide gel (SDS-PAGE), as well as analysis of brucella antigens reacting with IgG antibodies using immunoblotting with a set of specific S- and R-antiserums. Research has shown the similarity of protein profiles in brucella types which is absent in closely related bacteria, while immunoblot analysis has enabled type identification for brucella strains. Given this, the use of S-antiserums has enabled differentiation for strains of types В.melitensis, В.suis, В.abortus, and В.neotomae found in the S-form from strain types В.canis and В.canis found in the R-form. The practical importance of these methods was demonstrated given comparative study in connection with traditional methods of identification of 10 field strains of brucella taken from dogs. All detected strains have shown an identical protein profile picture and immunodominant antigens with reference strain B.canis RM6/66. This enabled confirmation of the strains as belonging to the type В.canis.
Comparative study into sensitivity to acid shock (pH below 4.0) is also being carried out on reference and vaccine strains of brucella that differ in their specific affiliation and pathogenic properties, as well as definition of the ability to increase resistance to acid shock following preliminary adaptation to soft рН 5.6 (a phenomenon known in literature as ATR [adaptive tolerance response]). The results obtained showed that the highly pathogenic types В.suis (biovar 1) and В.melitensis (biovar 3) are over 100 times more resistant to acid shock in comparison with low pathogenic type В.canis and the nonpathogenic for humans type В.neotomae.
The type specificities of brucella in its sensitivity to acid shock and ability to produce an ATR [adaptive tolerance response] must be considered when certifying brucella strains and vaccine strains, in particular where increased sensitivity to acid shock (pH 3.0-3.3) has been detected.
The head of the laboratory is N.A Zigangirova, Doctor of Biological Science.
The main line of research is the study of the biology of chlamydia - obligate intracellular pathogenic microorganisms of prokaryotic nature and infection pathology caused by them in addition to the development of new and furthering of existing methods for the indication of pathogens and diagnosis of chlamydial infection.
A direct microimmunofluorescent method for the determination of chlamydia using monoclonal antibodies has been developed and is widely used; microimmunofluorescent and microimmunoenzyme methods on slides for the determination of IgG, IgM and IgA antibodies have been developed and used. Commercial test systems based on these methods are being produced; new molecular and biological methods for determining the sensitivity of chlamydia to antibiotics are being developed.
A new concept in biochips is being developed for simultaneous definition of antibodies for a number of chlamydia agents. The biochip has the form of a glass slide with a depression housing a number of varying points with chlamydia antigens that obtain a specific tint in the case of a positive reaction.
In addition to the developments listed above, the laboratory constantly conducts diagnostic work analyzing patient material from infected cell cultures (gold standard).
Cell Microbiology Laboratory
The head of the laboratory is Russian Academy of Sciences corresponding member D.Y Logunov.
The laboratory was founded in 2012 on the basis of a decision of the President of the Russian Federation bringing together research work within the framework of priority lines of scientific development for the creation of medicinal and immuno-biological products that will define the development of preventative means (vaccines), treatments and diagnosis of pathogens causing chronic and socially significant infections.
The priority line of research for the laboratory is development of vaccines against pathogens causing illnesses of social significance, including development of tuberculosis vaccines.
Bacteria Genetics Laboratory
The director is G.I. Karataev, Doctor of Biological Science.
The laboratory was founded by academician V.D Timakov in 1964 for the purposes of studying the fundamental issues of genetics of microorganisms that are important for medical microbiology.
Since inception until 2000, the laboratory was led by Russian Academy of Medical Sciences academician Professor A.G Skavronskaya. Study of the molecular and genetic behavior of induced mutagens in bacteria and SOS-mutagenesis mechanisms is the main line of research. A range of important fundamental results have been obtained regarding the molecular mechanisms of DNA repair in pathogenic and nonpathogenic bacteria and the induction of mutations by DNA base analogs. A hybrid strain of S.typhimurium has been developed for more effective definition of mutagens and potential carcinogens in the Ames test. Since 1974, the laboratory has studied the fundamental mechanisms of pathogenicity and immunogenicity of brucellosis and tularemia pathogens, as a result of which genetically based methods for constructing attenuated bacteria for potential vaccine strains have been developed, and collections of virulent and attenuated mutations have been created.
The laboratory has been overseen by G.I. Aleshkin, Doctor of Medical Science since 2000. Under his leadership, studies have been carried out on the functioning of Y-family DNA polymerases in SOS mutagenesis and antimutagenesis in pathogenic and nonpathogenic bacteria, and work has begun on the molecular and genetic mechanisms of antimutagenesis connected with the functional interaction of the SOS system of mutagenesis and the regulatory FCS system of transport and utilization of carbohydrates in the bacterial cell. This research has enabled collection of priority genetic data regarding the appearance of carcinogens and mutagens, glucides, phage DNA and chromosomal DNA in the bacterial cell, and the specifics of expressions of genes of mobile genetic elements in pathogenic bacteria. Results have enabled commencement of study of the known behavior of obtaining bacteria E.coli, S.typhimurium and Y.pseudotuberculosis from genes of pathogenicity using intergeneric horizontal plasmid transfer and bacteriophages and the role of generatrice hybrids in the formation of genomes of new pathogenic microorganisms. Within the framework of this research, the possibility of lysogenization of pathogenic microorganisms of types Escherichia, Salmonella and Yersinia via the most widespread bacteriophages of the Microviridae family was described for the first time in the world.
Since 2012, under the guidance of Dr. G.I Karataev, the laboratory has researched the role of mobile genetic elements in the mutation and regulation of the virulent agent in whooping cough, B.pertussis. The mechanism of formation of persistent bacteria in the process of development of infection and its role in spreading disease is currently being studied. A test system is under development for laboratory diagnosis of whooping cough, which is a characteristically virulent bacterial population in varying stages of the infection process and a persistent agent in the human body.
The main line of applied research is the development and testing of a live vaccine for the prevention of whooping cough, based on the use of attenuated bacteria B.pertussis created in the laboratory. The live whooping cough vaccine obtained for intranasal administration is currently at the pre-clinical trial stage.