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The Journal of Eukaryotic Microbiology Nov 2016The virulence of various amoebic parasites has been correlated with the presence of electron-dense granules (EDGs) in the cytoplasm of trophozoites. Here, we report the...
The virulence of various amoebic parasites has been correlated with the presence of electron-dense granules (EDGs) in the cytoplasm of trophozoites. Here, we report the finding by transmission electron microscopy of a large number of EDGs in a recent culture of Acanthamoeba culbertsoni, isolated from a severe case of human keratitis. When this isolate was maintained in culture for 6 mo, the granules almost disappeared. However, after induction of mice brain lesions with the long-term cultured isolate, recovered amoebas had abundant EDGs. Trophozoites of the original isolate, or those recovered from experimental lesions, secreted EDGs into the medium when incubated with MDCK cells. To analyze a possible cytotoxic effect the conditioned medium was incubated with MDCK monolayers. After 5 h, the media containing EDGs produced opening of the tight junctions; at 24 h, cell viability was compromised, and at 48 h most of the cells were detached from the monolayer. In contrast, trophozoites in long-term cultures did not release EDGs to the medium during incubation with MDCK cells, and the corresponding conditioned medium did not have any effect on MDCK monolayers. Our observations further support the hypothesis that EDGs play a role in the cytopathogenic mechanisms of A. culbertsoni.
Topics: Acanthamoeba; Amebiasis; Animals; Humans; Keratitis; Male; Mice; Mice, Inbred BALB C; Microscopy, Electron, Transmission; Trophozoites; Virulence
PubMed: 27087641
DOI: 10.1111/jeu.12321 -
Experimental Parasitology Dec 2017Acanthamoeba culbertsoni trophozoites, previously isolated from a human keratitis case with severe intraocular damage, were maintained in axenic culture. Co-incubation...
Acanthamoeba culbertsoni isolated from a clinical case with intraocular dissemination: Structure and in vitro analysis of the interaction with hamster cornea and MDCK epithelial cell monolayers.
Acanthamoeba culbertsoni trophozoites, previously isolated from a human keratitis case with severe intraocular damage, were maintained in axenic culture. Co-incubation of amoebae with MDCK cell monolayers demonstrated an apparent preference of the amoebae to introduce themselves between the cells. The trophozoites appeared to cross the cell monolayer through the tight junctions, which resulted in decreased trans-epithelial resistance (TER) measurements. Unexpectedly, after co-incubation of amoebae with hamster corneas, we observed that the trophozoites were able to cross the different cell layers and reach the corneal stroma after only 12 h of interaction, in contrast to other Acanthamoeba species. These observations suggest that this A. culbertsoni isolate is particularly pathogenic. Further research with diverse methodologies needs to be performed to explain the unique behavior of this Acanthamoeba strain.
Topics: Acanthamoeba; Acanthamoeba Keratitis; Animals; Cornea; Cricetinae; Dogs; Epithelial Cells; Humans; Intercellular Junctions; Madin Darby Canine Kidney Cells; Microscopy, Electron, Scanning; Microscopy, Electron, Transmission; Microscopy, Fluorescence; Microscopy, Phase-Contrast; Trophozoites; Virulence
PubMed: 28974450
DOI: 10.1016/j.exppara.2017.09.018 -
Microorganisms Nov 2023Several species of genus are potential pathogens and etiological agents of several diseases. The pathogenic mechanisms carried out by these amoebae in different target...
Several species of genus are potential pathogens and etiological agents of several diseases. The pathogenic mechanisms carried out by these amoebae in different target tissues have been documented, evidencing the relevant role of contact-dependent mechanisms. With the purpose of describing the pathogenic processes carried out by these protozoans more precisely, we considered it important to determine the emission of extracellular vesicles (EVs) as part of the contact-independent pathogenicity mechanisms of , a highly pathogenic strain. Through transmission electronic microscopy (TEM) and nanoparticle tracking analysis (NTA), EVs were characterized. EVs showed lipid membrane and a size between 60 and 855 nm. The secretion of large vesicles was corroborated by confocal and TEM microscopy. The SDS-PAGE of EVs showed proteins of 45 to 200 kDa. Antigenic recognition was determined by Western Blot, and the internalization of EVs by trophozoites was observed through Dil-labeled EVs. In addition, some EVs biological characteristics were determined, such as proteolytic, hemolytic and COX activity. Furthermore, we highlighted the presence of leishmanolysin in trophozites and EVs. These results suggest that EVs are part of a contact-independent mechanism, which, together with contact-dependent ones, allow for a better understanding of the pathogenicity carried out by .
PubMed: 38004773
DOI: 10.3390/microorganisms11112762 -
Infection and Immunity Sep 2010Acanthamoeba culbertsoni is an opportunistic pathogen that causes granulomatous amoebic encephalitis (GAE), a chronic and often fatal disease of the central nervous...
Acanthamoeba culbertsoni is an opportunistic pathogen that causes granulomatous amoebic encephalitis (GAE), a chronic and often fatal disease of the central nervous system (CNS). A hallmark of GAE is the formation of granulomas around the amoebae. These cellular aggregates consist of microglia, macrophages, lymphocytes, and neutrophils, which produce a myriad of proinflammatory soluble factors. In the present study, it is demonstrated that A. culbertsoni secretes serine peptidases that degrade chemokines and cytokines produced by a mouse microglial cell line (BV-2 cells). Furthermore, soluble factors present in cocultures of A. culbertsoni and BV-2 cells, as well as in cocultures of A. culbertsoni and primary neonatal rat cerebral cortex microglia, induced apoptosis of these macrophage-like cells. Collectively, the results indicate that A. culbertsoni can apply a multiplicity of cell contact-independent modes to target macrophage-like cells that exert antiamoeba activities in the CNS.
Topics: Acanthamoeba; Animals; Apoptosis; Brain; Cell Line; Chemokines; Cytokines; Immune Evasion; Mice; Microglia; RNA, Messenger; Rats; Serine Proteases
PubMed: 20605979
DOI: 10.1128/IAI.00047-10 -
Antioxidants (Basel, Switzerland) Dec 2023is a ubiquitous genus of amoebae that can act as opportunistic parasites in both humans and animals, causing a variety of ocular, nervous and dermal pathologies....
is a ubiquitous genus of amoebae that can act as opportunistic parasites in both humans and animals, causing a variety of ocular, nervous and dermal pathologies. Despite advances in therapy, the management of patients with infections remains a challenge for health services. Therefore, there is a need to search for new active substances against Acanthamoebae. In the present study, we evaluated the amoebicidal activity of nitroxoline against the trophozoite and cyst stages of six different strains of . The strain showed the lowest IC value in the trophozoite stage (0.69 ± 0.01 µM), while the strain L-10 showed the lowest IC value in the cyst stage (0.11 ± 0.03 µM). In addition, nitroxoline induced in treated trophozoites of features compatibles with apoptosis and autophagy pathways, including chromatin condensation, mitochondrial malfunction, oxidative stress, changes in cell permeability and the formation of autophagic vacuoles. Furthermore, proteomic analysis of the effect of nitroxoline on trophozoites revealed that this antibiotic induced the overexpression and the downregulation of proteins involved in the apoptotic process and in metabolic and biosynthesis pathways.
PubMed: 38136200
DOI: 10.3390/antiox12122081 -
Experimental Parasitology Sep 2018Amoebae from the genus Acanthamoeba are facultative pathogens of humans and other animals. In humans they most frequently infect the eye causing a sight threatening...
Amoebae from the genus Acanthamoeba are facultative pathogens of humans and other animals. In humans they most frequently infect the eye causing a sight threatening infection known as Acanthamoeba keratitis (AK), and also cause an often fatal encephalitis (GAE). A mannose-binding protein (MBP) has been identified as being important for Acanthamoeba infection especially in AK. This lectin has previously been characterized from Acanthamoeba castellanii as consisting of multiple 130 kDa subunits. MBP expression correlates with pathogenic potential and is expressed in a number of Acanthamoeba species. Here we report the purification of a similar lectin from Acanthamoeba culbertsoni and the production of a monoclonal antibody to it. The A. culbertsoni MBP was isolated by affinity chromatography using α-D-mannose agarose and has an apparent molecular weight of 83 kDa. The monoclonal antibody is an IgM that is useful in both western blots and immunofluorescence. We expect that this antibody will be useful in the study of the pathology of A. culbertsoni and in its identification in clinical samples.
Topics: Acanthamoeba; Acanthamoeba Keratitis; Animals; Antibodies, Monoclonal; Antibodies, Protozoan; Antigens, Protozoan; Blotting, Western; Electrophoresis, Polyacrylamide Gel; Enzyme-Linked Immunosorbent Assay; Hybridomas; Immune Sera; Immunoglobulin Isotypes; Immunohistochemistry; Mannose-Binding Lectin; Mice; Mice, Inbred BALB C; Protozoan Proteins
PubMed: 30031120
DOI: 10.1016/j.exppara.2018.07.009 -
Experimental Parasitology Sep 2010Acanthamoeba are free-living amoebae found in most environments that can cause brain and corneal infections. To infect humans, these pathogens must interact with host...
Acanthamoeba are free-living amoebae found in most environments that can cause brain and corneal infections. To infect humans, these pathogens must interact with host cells and the extracellular matrix (ECM). In order to define the mode by which amoebae recognize ECM components and process this recognition, we analyzed Acanthamoeba culbertsoni attachment and invasion, respectively, on collagen I and laminin-1 and on tridimensional collagen I and matrigel matrices. We determined that amoebae surface proteins are involved in adhesion, that exogenous sugars can decrease adhesion and invasion, and that adhesion and invasion are dependent on microfilament reorganization. In addition, we determined the role of serine- and metallo-proteases on invasion and found that adhesion was blocked when amoebae were treated with a metallo-protease inhibitor. Collectively, these results suggest that adhesion and invasion are protease- and microfilament-dependent events in which amoebic surface proteins play a pivotal role.
Topics: Acanthamoeba; Bridged Bicyclo Compounds, Heterocyclic; Carbohydrates; Cell Adhesion; Collagen; Colorimetry; Extracellular Matrix; Laminin; Peptide Hydrolases; Periodic Acid; Thiazolidines; Trypsin
PubMed: 19698710
DOI: 10.1016/j.exppara.2009.08.004 -
Kisaengch'unghak Chapchi. the Korean... Dec 1990Studies on pathogenic free-living amoebae performed in Korea were briefly reviewed. One strain of Naegleria fowleri was isolated from the sewage, and 3 strains of... (Review)
Review
Studies on pathogenic free-living amoebae performed in Korea were briefly reviewed. One strain of Naegleria fowleri was isolated from the sewage, and 3 strains of Acanthamoeba culbertsoni from a reservoir and the gill of fish. They were identified by morphological characteristics. Three strains among the 4 were experimentally proved pathogenic to cause primary amoebic meningoencephalitis in mice. The virulence of N. fowleri depended upon various factors such as strain, weight and sex of mice, and number of the inoculated amoebae. A. culbertsoni was found to retain cytolytic activities which were related to the pathogenicity. Also these amoebae were demonstrated to harbour the activities of acid phosphatase, peroxidase and ATPase. There is one case record of systemic Acanthamoeba infection in Korea, proved at autopsy. Cellular or humoral responses to these amoebae have been studied in vitro or in vivo. Immunization could reduce the mortality of experimentally infected mice.
Topics: Acanthamoeba; Amebiasis; Animals; Naegleria fowleri
PubMed: 2133420
DOI: 10.3347/kjp.1990.28.suppl.29 -
The Korean Journal of Parasitology Mar 2006Free-living amoebae of the genus Acanthamoeba are causative agents of granulomatous amebic encephalitis and amebic keratitis. Because the virulence of Acanthamoeba... (Comparative Study)
Comparative Study
Free-living amoebae of the genus Acanthamoeba are causative agents of granulomatous amebic encephalitis and amebic keratitis. Because the virulence of Acanthamoeba culbertsoni cultured in the laboratory is restored by consecutive brain passages, we examined the genes induced in mouse brain-passaged A. culbertsoni by differential display reverse transcriptase polymerase chain reaction (DDRT-PCR). Enhanced A. culbertsoni virulence was observed during the second mouse brain passage, i.e., infected mouse mortality increased from 5% to 70%. Ten cDNAs induced during mouse brain passage were identified by DDRT-PCR and this was confirmed by northern blot analysis. BlastX searches of these cDNAs indicated the upregulations of genes encoding predictive NADH-dehydrogenase, proteasomal ATPase, and GDP-mannose pyrophosphorylase B, which have previously been reported to be associated with A. culbertsoni virulence factors.
Topics: Acanthamoeba; Amebiasis; Animals; Blotting, Northern; Brain; Cloning, Molecular; DNA, Complementary; DNA, Protozoan; Gene Expression Profiling; Gene Expression Regulation; Genes, Protozoan; Mice; Mice, Inbred ICR; Molecular Sequence Data; Reverse Transcriptase Polymerase Chain Reaction; Serial Passage; Up-Regulation; Virulence
PubMed: 16514277
DOI: 10.3347/kjp.2006.44.1.15 -
Pathogens (Basel, Switzerland) Nov 2023Few studies have been conducted in the cooling systems of power plants; they have focused on , leaving a gap in the knowledge of other pathogenic free-living amoebae in...
Few studies have been conducted in the cooling systems of power plants; they have focused on , leaving a gap in the knowledge of other pathogenic free-living amoebae in this environment. The objective of this study was to determine the occurrence of saline-tolerant pathogenic in a geothermal power plant. The identification of isolated amoebae at genus level was carried out, observing their morphological characteristics; the determination of genotype and species of was performed via molecular biology (PCR). Water temperature ranged from 18 to 43 °C and conductivity from 4.0 × 10 to 8.7 × 10 μS/cm; this last value was greater than the seawater value. Only five amoeba genera were found. was in all the sampling sites, showing high saline tolerance. The high temperature, but mainly high conductivity, were the environmental conditions that determined the presence of pathogenic free-living amoebae in the hot water. All the strains of killed the mice, having a mortality of 40 to 100%. genotypes T10 and T5 were identified, T10 is rarely isolated from the environment, while T5 is more frequent. This is the first time that genotypes T5 and T10 have been reported in the environment in Mexico.
PubMed: 38003827
DOI: 10.3390/pathogens12111363