Vojenské Zdravotnické Listy, 2002 (vol. 71), Suppl.1
INTRACELLULAR BACTERIAL PATHOGENS: FROM STRUCTURE TO PATHOGENICITY
Jiří Stulík
MMSL 2002, 71(S1):1-3 
FROM THE BEGINNING...
Aleš Macela, Jiří Stulík
MMSL 2002, 71(S1):4
Since 1972 the former Institute for Immunology of Purkyne Military Medical Academy has been engaged in the study of pathogenesis of severe viral, rickettsial and bacterial infections. In the late eighties this Institute built laboratory for the analysis of complex protein mixtures originating from prokaryotic and eukaryotic cells that was based on the application of high-resolution two-dimensional gel electrophoresis (2-DE). Furthermore, the complete proteome "technology including computer-assisted 2-DE gel evaluation and MALDI-TOF MS for protein identification, was successfully implemented in 1999. In the year 2000 the Institute launched the tight...
AN ATTEMPT TO DISCRIMINATE BORRELIA BURGDORFERI SENSU LATO COMPLEX MICROBES ACCORDING TO THE PROTEIN PROFILE
Věra Neubauerová, Aleš Macela, Jiří Stulík, Jana Havlasová, Lenka Hernychová, Zuzana Kročová, Jan Kopecký, Jitka Pechová
MMSL 2002, 71(S1):5-8
Lyme borreliosis (LB), the most common vector-borne disease in Europe, North America and parts of Asia, is caused by the tick-bome Spirochete Borrelia species. In 1982, the bacterium that causes LB was first isolated by Willy Burgdorfer and colleagues from the hard tick Ixodes dammini collected On Long Island, N. Y. [1]. The isolate was subsequently identified as a new species of the genus Borrelia and was named Borrelia burgdorferi in 1984 [2]. In the past years classification of Lyme disease borreliae has been investigated intenSively. More than 20 Species have been identified within the genus Borrelia...
IDENTIFICATION OF FRANCISELLA TULARENSIS PROTEINS
Irena Kasalová, Lenka Hernychová, Věra Neubauerová, Jiří Stulík, Jana Havlasová, Aleš Macela
MMSL 2002, 71(S1):9-10
Francisella tularensis is the Gram - negative, facultative intracellular bacterium which is able to cause tularemia. It can survive in macrophages or hepathocytes. Francisella tularensis is devided into four subspecies F. tularensis subsp. tularensis, F tularensis subsp. palearctica, F. tularensis subsp. mediaasiatica, F tularensis subsp. palearctica japonica. Live attenuated Vaccine Strain (LVS) Selected from a mixture of attenuated strains is used as an experimental model for human infection in mıce....
IMMUNO-REACTIVE PROTEINS OF FRANCISELLA TULARENSIS
Jana Havlasová, Jiří Stulík, Lenka Hernychová
MMSL 2002, 71(S1):11-16
Francisella tularensis is the ethiological agent of tularemia, an infection of humans and other mammals. The laboratory diagnosis of human tularemia is based on serological tests. However, there is a lack of information about the Francisella tularensis antigens inducing antibody response to date. Except for the complex antigen preparations as sonicated whole bacteria [Viljanen 1983], only the Francisella lipopolysaccharide [Carlsson 1979] and the partially purified fraction containing outer membrane antigens [Bevanger 1988] have been described as immunogens suitable for diagnostic purposes. Proteome technology, based on two-dimensional...
FUNCTIONAL PROFILE OF FRANCISELLA TULARENSIS PROTEINS
Lenka Hernychová, Aleš Macela, Jiří Stulík
MMSL 2002, 71(S1):17-19
In spite of the enormous quantity of information from genomic and proteomic analysis the data do not mirror precisely in vivo situation in respect to functional profile of cellular system studied. Generally, the function of molecular entities is strictly associated with their tertiary structure, subcellular localization inside individual cell compartments andbasically on the consensual sequences, which compose the functional domains of protein molecule. Moreover, the metabolic and signaling cellular cascades rely on the protein-protein interactions, which are alsoout of the scope of, so called “expression proteomics”. To overcome...
AN IN VITRO MODEL OF FRANCISELLA TULARENSIS LVS INFECTION
Monika Černá, Aleš Macela, Jan Krejsek
MMSL 2002, 71(S1):20-21
Francisella tularensis live vaccine strain (LVS) is a facultative intracellular bacterium, which causes lethal disease in several mouse strains (Fortier et al. 1991). A huge body of information dealing with the mechanisms of Francisella tularensis infection of various mouse cell lines is available (Macela 1980, Fortier et al. 1992). In this study we used in vitro infection of murine macrophage-like cell line J774 for the standardization of another in vitro model, which could be more universal for the studies of bacteria — host pathogen interactions, more precise assignment of early stages of primary infection of host...
PHENOTYPIC PROFILE OF PHAGOCYTIC CELLS J774.2 INFECTED WITH FRANCISELLA TULARENSIS LVS MICROBES
Martina Loudová, Monika Černá, Jan Krejsek, Aleš Macela
MMSL 2002, 71(S1):22-25
The Overall aim Of this study was to analyze the phenotypic changes of the host cells that can occur during the primary interaction of Francisella tularensis microbes with professional phagocytes. The cell line J774.2, derived from female BALB/c mouse (ECACC, No. 85011428), Was used as a model of host cell, which can represent first line of defense in host tissues infected with intracellular bacterial pathogens. Francisella tularensis live vaccine strain (Francisella tularensis LVS, ATCC 29684, American type culture collection, ManasSas, .Va, USA) Was used for infection in vitro. Detection and identification of the...
BACTERIAL EFFECTORS AND HOST MOLECULAR TARGETS WHICH CONTROL THE ORIGIN, MATURATION, AND FINAL DESTINY OF PHAGOCYTIC VACUOLE CONTAINING THE INTRACELLULAR BACTERIAL PATHOGEN
Stanislava Porkertová, Hana Kovářová
MMSL 2002, 71(S1):26-28
Intracellular bacteria and parasites have evolved many different strategies to elude host defense mechanisms. Although some of the pathogens including Listeria, Shigella, and Rickettsia escape into the cytoplasm to avoid lysosomal digestion, the major group is capable to adapt to intracellular environment and reside in membrane bound vesicles — phagosomes. The phagosomes containing intracellular parasites can arrest in the early (Mycobacterium spp., Ehrlichia chaffeensis) or late (Leishmania) endosomal stage of maturation and even mature to phagolysosome and adapt to this environment (Coxiella burnetii)....
F. TULARENSIS INFECTION-INDUCED CELL CYCLE BLOCK AND INDUCTION OF APOPTOSIS
Roman Hrstka, Bořivoj Vojtěšek
MMSL 2002, 71(S1):29-31
Generally, macrophages initiate the innate immune response by recognizing pathogens, phagocytosing them and secreting inflammatory mediators. However, intracellular bacterial pathogens have developed plenty of strategies to influence processes in eucaryotic host cells to establish appropriate conditions for their own growth and proliferation.Francisella tularensis is a highly virulent intracellular pathogen responsible for tularemia. This bacterium is capable of infecting many mamalian species and various types of cells, but little is known about the mechanisms of survival and interactions with host cells....
CYTOKINE AND CHEMOKINE MRNA EXPRESSION DURING IN VITRO INFECTION OF PHAGOCYTIC CELLS — CURRENT STATE OF STUDY
Blanka Hartmanová, Zdeněk Fiedler
MMSL 2002, 71(S1):32-35
The aim of this work is to analyze the expression of eukaryotic genome of the phagocytic cell after infection with microorganism, in particular after infection with Francisella tularensis. Mouse macrophage cell line J774 is used in experiments as a model of phagocytic cell. The first part of activities which should lead to reach the aim was selecting of suitable markers of the infection. The expression of these markers should be affected by the infection of phagocytes. We chose some cytokines, chemokines and also enzymes....
LOCAL IN VIVO HOST RESPONSES TO INFECTION OF MICE WITH FRANCISELLA TULARENSIS LVS
Zuzana Kročová
MMSL 2002, 71(S1):36-38
Francisella tularensis is the causative agent of the disease tularemia. Inhalation of infected dust leads to the respiratory form of the disease, while transmission by arthropods or direct contact with an infected animal causes the ulceroglandular form of the disease. Lethality in mouse model is much more dependent on the route of infection than on the genetic background of mice. LD50 after i.d. infection with F tularensis LVS at both — either susceptible (BALB/c) or resistant (C57Bl/ 10) mouse strain is higher than 1x107 microbes per mouse [1]. On the Other hand, LD50 after aerogenic infection is 3x101...
MICROBIAL PROTEINS: FROM STRUCTURE TO PATHOGENICITY — GROEL AS AN EXAMPLE
Aleš Macela, Lenka Hernychová, Jana Havlasová, Jiří Stulík
MMSL 2002, 71(S1):39-43
Genomic and proteomic analysis bring to biomedical labs qualitatively new technologies, which enable the complex look to genes and gene-related products — proteins. In contrary to genes, which number is definitive for given living system, the number of protein species varied substantially according the immediate living conditions of the system studied. Moreover, most of the proteins have the function associated with the tertiary structure of their molecule, e.g. with the space orientation so called folding and intracellular compartment where are they located. In in vitro systems the process of folding is inefficient and only minority of...