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Dive into the research topics where Zuzana Kročová is active.

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Featured researches published by Zuzana Kročová.


Infection and Immunity | 2003

An Attenuated Strain of the Facultative Intracellular Bacterium Francisella tularensis Can Escape the Phagosome of Monocytic Cells

Igor Golovliov; Vladimir Baranov; Zuzana Kročová; Hana Kovarova; Anders Sjöstedt

ABSTRACT The facultative intracellular bacterium Francisella tularensis is a highly virulent and contagious organism, and little is known about its intracellular survival mechanisms. We studied the intracellular localization of the attenuated human vaccine strain, F. tularensis LVS, in adherent mouse peritoneal cells, in mouse macrophage-like cell line J774A.1, and in human macrophage cell line THP-1. Confocal microscopy of infected J774A.1 cells indicated that during the first hour of infection the bacteria colocalized with the late endosomal-lysosomal glycoprotein LAMP-1, but within 3 h this colocalization decreased significantly from approximately 60% to 30%. Transmission electron microscopy revealed that >90% of bacteria were not enclosed by a phagosomal membrane after 2 h of infection, and some bacteria were in vacuoles that were only partially surrounded by a limiting membrane. Similar findings were obtained with all three host cell types. Immunoelectron microscopy performed with an F. tularensis LVS-specific polyclonal rabbit antiserum showed that the antiserum stained a thick, evenly distributed capsule-like material in bacteria grown in broth. In contrast, intracellular F. tularensis LVS cells were only marginally stained with this antiserum. Instead, most of the immunoreactive material was diffusely localized in the phagosomes or was associated with the phagosomal membrane. Our findings indicate that F. tularensis LVS is able to escape from the phagosomes of macrophages via a mechanism that may involve degradation of the phagosomal membrane.


Toxicology in Vitro | 2000

The immunomodulatory effect(s) of lead and cadmium on the cells of immune system in vitro

Zuzana Kročová; Aleš Macela; Michal Kroca; Lenka Hernychová

A number of studies documented that the heavy metals are not only toxic for the organisms but they may modulate immune responses. The immunomodulatory activity was proved in several in vivo and in vitro model systems. In the current study, immunomodulatory activities of lead and cadmium are presented. The viability of both lymphocytes and macrophages was affected by heavy metals in a dose- and time-dependent manner. In the case of lead, the depression of N-oxide production closely correlated with increased blast transformation of spleen cells induced by concanavalin A (ConA). On the contrary, cadmium suppressed the production of N-oxides but stimulated significantly the proliferation of spleen cells. The production of cytokines by lymphocytes and macrophages was dependent on the in vitro model used. Generally, the treatment of macrophages with lead results in disregulation of the production of proinflammatory cytokines [tumour necrosis factor alpha (TNF-alpha), interleukin 1alpha (IL-1alpha) and interleukin 6 (IL-6)] and preferential production of Th1 type of cytokines (IFN-gamma and IL-2). Cadmium seemed to trigger the Th2 cytokine regulatory pathway [interleukin 4 (IL-4), interleukin 10 (IL-10)]. The results suggest the metal-induced changes in immunoregulatory mechanism of host with potentially severe clinical consequences.


Pharmacology & Toxicology | 2001

Toxic effects of sarin in rats at three months following single or repeated low-level inhalation exposure

Jiří Kassa; Miroslav Pecka; Miloš Tichý; Jiří Bajgar; Marie Koupilová; Josef Herink; Zuzana Kročová

Male albino Wistar rats were once or repeatedly exposed to three various low concentrations of sarin for 60 min. in the inhalation chamber. The clinical status of control as well as sarin-poisoned rats was tested 3 months after exposure to sarin using biochemical, haematological, neurophysiological, behavioural and immunotoxicological methods. While biochemical and haematological parameters, including the activities of cholinesterases in erythrocytes, plasma and various organs (brain, diaphragm), did not differ from the control values regardless of the sarin concentration used, few signs of sarin-induced neurotoxicity and immunotoxicity in sarin-poisoned rats were demonstrated. This was especially true when the single exposure of rats to non-convulsive symptomatic concentration and repeated exposure of rats to clinically asymptomatic concentration of sarin was used. In rats repeatedly poisoned with clinically asymptomatic concentrations of sarin, the alteration of the gait characterized by ataxia, the increase in the stereotyped behaviour, the increase in the excitability of the central nervous system following the administration of the convulsive drug pentamethylenetetrazol were observed. In rats poisoned with non-convulsive symptomatic concentration of sarin, the subtle supression of spontaneous, as well as lipopolysaccharides-stimulated, proliferation of spleen lymphocytes and the bactericidal activity of peritoneal macrophages was primarily observed besides the signs of neurotoxicity. Our findings confirm that both non-convulsive symptomatic and clinically asymptomatic concentrations of sarin can only cause very few, subtle long-term signs of neurotoxicity and immunotoxicity in sarin-poisoned rats when the rats were exposed to asymptomatic sarin concentrations repeatedly.


Toxicology | 2003

Low-level sarin-induced alteration of immune system reaction in inbred BALB/c mice

Jiří Kassa; Zuzana Kročová; Lucie Ševelová; Irena Kasalová; Věra Neubauerová

To study the influence of low-level sarin inhalation exposure on immune functions, inbred BALB/c mice were exposed to low concentrations of sarin for 60 min in the inhalation chamber. Two concentrations of sarin were chosen-asymptomatic concentration (LEVEL 1) and non-convulsive symptomatic concentration (LEVEL 2). The evaluation of immune functions was carried out using phenotyping of CD3 (T-lymphocytes), CD4 (helper T-lymphocytes), CD8 (cytotoxic T-lymphocytes) and CD19 cells (B-lymphocytes) in the lungs, blood and spleen, lymphoproliferation of spleen cells stimulated in vitro by various mitogens (concanavalin A, lipopolysaccharides), phagocyte activity of peritoneal and alveolar macrophages, production of N-oxides by peritoneal macrophages and the measurement of the natural killer cell activity at 1 week following sarin exposure. The results were compared to the values obtained from control mice exposed to pure air instead of sarin. The results indicate that not only symptomatic but also asymptomatic dose of sarin is able to alter the reaction of immune system at 1 week following exposure to sarin. While the number of CD3 cells in the lungs was slightly decreased, an increase in CD19 cells was observed especially in the lungs and blood. The reduced proportion of T-lymphocytes is caused by decay of CD4 positive T-cells. Lymphoproliferation was significantly decreased regardless of the mitogen and sarin concentration used. The production of N-oxides by peritoneal macrophages was stimulated after exposure to LEVEL 2 of sarin whereas their ability to phagocyte the microbes was increased after exposure to LEVEL 1. The natural killer cell activity was significantly higher in the case of inhalation exposure of mice to LEVEL 2 of sarin. Thus, not only organophosphorus insecticides but also nerve agents such as sarin are able to alter immune functions even at a dose that does not cause clinically manifested intoxication following the inhalation exposure. Nevertheless, the alteration of immune functions following the inhalation exposure to a symptomatic concentration of sarin seems to be more pronounced.


Proteomics | 2002

Proteome study of Francisella tularensis live vaccine strain-containing phagosome in Bcg/Nramp1 congenic macrophages: resistant allele contributes to permissive environment and susceptibility to infection.

Hana Kovářová; Petr Halada; Petr Man; Igor Golovliov; Zuzana Kročová; Josef Špaček; Stanislava Porkertová; Radka Nečasová

The phagocytosis of pathogens by macrophages classically initiates maturation of the phagosome that involves a dynamic exchange of phagosomal components with intracellular compartments of the endocytic pathway. The intracellular microorganisms have developed sophisticated mechanisms to sense environmental conditions and respond to them by phenotypic alterations that ensure their adaptation, survival and proliferation inside the cell. They have learned also to utilise host cellular components to ensure own survival. Recent results suggest that the Bcg locus/Nramp1 gene (natural resistance‐associated macrophage protein 1) controls natural resistance to infection by Francisella tularensis LVS (live vaccine strain) and its effect is opposite to that observed for other Bcg/Nramp1‐controlled pathogens such as several mycobacterial species, Leischmania donovani, and Salmonella typhimurium. In the case of F. tularensis LVS infection, the mutant allele of the Bcg locus (Bcgs/Nramp1s) is associated with natural resistance and, inversely, the wild type allele (Bcgr/Nramp1r) confers susceptibility. To determine whether differential allelic expression of the Bcg locus/Nramp1 gene modifies the composition of F. tularensis LVS‐containing phagosomes (FCP), we have utilised an approach where we isolated FCP from infected Bcg congenic B10R (Bcgr/Nramp1r) and B10S (Bcgs/Nramp1s) macrophages of susceptible and resistant phenotype, respectively. Comparative proteomic analysis of the two phagosomal compartments with subsequent mass spectrometric analysis allowed identification of several proteins typical for FCP from B10R macrophages. They include a bacterial hypothetical 23 kDa protein, 60 kDa chaperonin GroEL, and host putative proteins that appeared to be mitochondrial ATP synthase β‐chain and NADH‐ubiquinone oxidoreductase based on high cross‐species homology. High abundance of the hypothetical 23 kDa protein correlates with the susceptible phenotype and, possibly, pathogenicity of F. tularensis LVS. The results demonstrate that F. tularensis LVS can exploit ion transport function of Bcg/Nramp1 to its own advantage.


Folia Microbiologica | 1999

Production of stress-inducible form of heat-shock protein 70 in mouse peritoneal adherent cells after in vivo infection by Francisella tularensis.

Jiří Stulík; Lenka Hernychová; Aleš Macela; Zuzana Kročová; Michal Kroca

Heat-shock proteins (hsp) are ubiquitously produced molecules which participate in the protection of cells from environmental perturbation. Moreover, the members of the heat-shock protein 60 (hsp60) and 70 (hsp70) families play an important role in pathogen-host interactions. We studiedin vivo production of the 70-kDa heat-shock proteins in the extract of peritoneal exudate cells (PEC) from mice injected intraperitoneally with an attenuated vaccine strain (LVS) ofFrancisella tularensis. We found a differential production of a highly stress-inducible member of the hsp70 family, designated hsp72, in three inbred strains of mice exhibiting either resistance or susceptibility toF. tularensis LVS infection. Whereas in tularemia-resistant mice hsp72 was even expressed in PEC without injection of bacteria and its production further increased on day 3 and slowly declined on days 5 and 7 after injection, in susceptible mice hsp72 production was highly inducble and restricted only to day 3 afterin vivo infection. Further analysis of hsp72 expression revealed intracellular hsp72 accumulation and its preferential production by peritoneal adherent cells.


Inhalation Toxicology | 2004

The Alteration of Immune Reactions in Inbred BALB/c Mice Following Low-Level Sarin Inhalation Exposure

Jiří Kassa; Zuzana Kročová; Lucie Ševelová; Valerie Sheshko; Irena Kasalová; Věra Neubauerová

To study the influence of low-level sarin inhalation exposure on immune functions, inbred BALB/c mice were exposed to low concentrations of sarin for 60 min in the inhalation chamber. The evaluation of immune functions was carried out using phenotyping of CD3 (T lymphocytes), CD4 (helper T lymphocytes), CD8 (cytotoxic T lymphocytes), and CD19 cells (B lymphocytes) in the lungs, blood, and spleen, lymphoproliferation of spleen cells stimulated in vitro by various mitogens (concanavalin A, lipopolysaccharides), phagocyte activity of peritoneal and alveolar macrophages, production of N-oxides by peritoneal macrophages, and the measurement of the natural killer cell activity at 1 wk following sarin exposure. The results were compared to the values obtained from control mice exposed to pure air instead of sarin. The results indicate that low doses of sarin are able to alter the reaction of immune system at one week following exposure to sarin. While the numbers of CD3 cells in the lungs, blood, and spleen were slightly decreased, an increase in CD19 cells was observed, especially in the lungs and blood. The reduced proportion of T lymphocytes is caused by decay of CD4-positive T cells. Lymphoproliferation was significantly decreased regardless of the mitogen and sarin concentration used. The production of N-oxides by peritoneal macrophages was stimulated after exposure to the highest dose of sarin, whereas their ability to phagocytize the microbes was increased after exposure to the lowest dose of sarin. The natural killer cell activity was significantly higher in the case of inhalation exposure of mice to the highest level of sarin. Thus, not only organophosphorus insecticides but also nerve agents such as sarin are able to alter immune functions even at a dose that does not cause clinically manifested disruption of cholinergic nervous system in the case of inhalation exposure. Nevertheless, the alteration of immune functions following the inhalation exposure to a symptomatic concentration of sarin seems to be more pronounced.


Archive | 2000

Measurement of Biological Aerosols in an Aerosol Chamber

Zuzana Kročová; J. Krejci; Michal Kroca; Aleš Macela

A standard system for the aerosol exposure of mice with viable Francisella tularensis live vaccine strain (LVS) in an aerosol chamber was developed. The pneumatic nebulizer and the dynamic “nose only” aerosol chamber for exposure of 12 mice were used. Pressure of the orifice and flow rate through the atomizer, flow rate of the air, temperature, and humidity in the chamber were measured. Density, viscosity, consumption of the aerosolized suspension, and the number of cultivable bacteria in the suspension were measured. The correlation between the technical parameters and the number of the cultivable bacteria in the initial suspension, aerosol, and the murine lungs were determined.


Fems Immunology and Medical Microbiology | 1996

The immune response against Francisella tularensis live vaccine strain in Lpsn and Lpsd mice

Aleš Macela; Jiri Stulik; Lenka Hernychová; Michal Kroca; Zuzana Kročová; Hana Kovarova


Basic & Clinical Pharmacology & Toxicology | 2004

The influence of single or repeated low-level sarin exposure on immune functions of inbred BALB/c mice.

Jiří Kassa; Zuzana Kročová; Lucie Ševelová; Irena Kasalová; Věra Neubauerová

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Aleš Macela

Military Medical Academy

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Michal Kroca

Military Medical Academy

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Hana Kovarova

Military Medical Academy

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Jiří Kassa

Military Medical Academy

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Jiří Stulík

Military Medical Academy

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L. Hemychova

Military Medical Academy

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