RESUMO
Amoebiasis is a disease caused by Entamoeba histolytica, affecting the large intestine of humans and occasionally leading to extra-intestinal lesions. Entamoeba dispar is another amoeba species considered commensal, although it has been identified in patients presenting with dysenteric and nondysenteric colitis, as well as amoebic liver abscess. Amoebic virulence factors are essential for the invasion and development of lesions. There is evidence showing that the association of enterobacteria with trophozoites contributes to increased gene expression of amoebic virulence factors. Enteropathogenic Escherichia coli is an important bacterium causing diarrhea, with high incidence rates in the world population, allowing it to interact with Entamoeba sp. in the same host. In this context, this study aims to evaluate the influence of enteropathogenic Escherichia coli on ACFN and ADO Entamoeba dispar strains by quantifying the gene expression of virulence factors, including galactose/N-acetyl-D-galactosamine-binding lectin, cysteine proteinase 2, and amoebapores A and C. Additionally, the study assesses the progression and morphological aspect of amoebic liver abscess and the profile of inflammatory cells. Our results demonstrated that the interaction between EPEC and ACFN Entamoeba dispar strains was able to increase the gene expression of virulence factors, as well as the lesion area and the activity of the inflammatory infiltrate. However, the association with the ADO strain did not influence the gene expression of virulence factors. Together, our findings indicate that the interaction between EPEC, ACFN, and ADO Entamoeba dispar strains resulted in differences in vitro and in vivo gene expression of Gal/GalNAc-binding lectin and CP2, in enzymatic activities of MPO, NAG, and EPO, and consequently, in the ability to cause lesions.
Assuntos
Entamoeba , Escherichia coli Enteropatogênica , Fatores de Virulência , Escherichia coli Enteropatogênica/patogenicidade , Escherichia coli Enteropatogênica/genética , Entamoeba/patogenicidade , Entamoeba/genética , Entamoeba/fisiologia , Fatores de Virulência/genética , Virulência , Animais , Camundongos , Abscesso Hepático Amebiano/parasitologia , Entamebíase/parasitologia , Humanos , Expressão GênicaRESUMO
Entamoeba histolytica is a protozoan-pathogen-causing amoebic liver abscess (ALA). After amoeba establishment in the liver, it causes abundant infiltrate of neutrophils. Liver tissue damage by neutrophils results in part from anti-amoebic oxidative intermediates, including reactive oxygen species (ROS), reactive nitrogen species (RNS), and hypochlorous acid (HOCl), derived from the myeloperoxidase (MPO) enzyme. Ascorbic acid (ASC) is an antioxidant that acts as a scavenger for ROS and NOS-derived free radicals. No previous information regarding the effect of ASC concerning the participation of MPO in an experimental model of ALA in hamsters has been reported. Thus, the aim of the present work was to analyze the effect of ASC on acute ALA development and to measure the activity and gene expression of the MPO enzyme. Hamsters were treated with ASC (800 mg/kg) and then intrahepatically inoculated with E. histolytica trophozoites. Animals were sacrificed at 3, 6, and 12 h post-inoculation (p.i.), and liver samples were collected. The percentage of lesions, amoeba in situ count, MPO activity, and mpo gene expression were ascertained. Compared to ALA hamsters without ASC treatment as the control group (CT), the ALA group treated with ASC had a significant decrease in liver lesions (all p.i. hours) and viable amoeba count (12 h p.i.) and an increase in MPO activity (12 h p.i.) and mpo gene expression (6 h/12 h p.i.). These data suggest that ASC ameliorated liver damage caused by oxidizing products via modulation of mpo expression and activity.
Assuntos
Ácido Ascórbico , Abscesso Hepático Amebiano , Peroxidase , Animais , Ácido Ascórbico/farmacologia , Cricetinae , Entamoeba histolytica/patogenicidade , Abscesso Hepático Amebiano/tratamento farmacológico , Oxirredução , Estresse Oxidativo , Peroxidase/metabolismo , Espécies Reativas de OxigênioRESUMO
Entamoeba histolytica causes amoebic liver abscess (ALA) in humans. The injury of target cells by E. histolytica includes processes controlled by the ubiquitin Ehub. Previously, we found immunodominance of Ehub glycan moieties using immunized rabbits. In this work, we analysed dominance of antibodies to the glycoprotein Ehub in the sera from 52 patients with ALA. Controls were sera from 20 healthy people living in endemic areas with a high seroprevalence of antibodies to amoebas, and 20 patients with alcoholic hepatitis (AH) to rule out the cross-reaction of Ehub with autoantibodies induced by liver damage. Antigens were trophozoite extract, glycoprotein Ehub and the recombinant protein E. histolytica recombinant ubiquitin (rEhub). The sera from healthy volunteers and patients with AH do not have antibodies to glycoprotein Ehub. Surprisingly, only the antibodies from patients with ALA recognized the glycoprotein Ehub, and some sera gave a faint reaction with the recombinant protein, especially because evolutionarily, the ubiquitin is conserved between species. This is the first report demonstrating that antibodies to ubiquitin Ehub are induced exclusively in patients with invasive amoebiasis, and the antibody response is mainly to the glycoprotein, indicating glycans are immunodominant. Inhibitors of the Ehub glycans could be potential treatment for amoebiasis by selectively damaging trophozoites.
Assuntos
Amebíase , Disenteria Amebiana , Entamoeba histolytica , Amebíase/tratamento farmacológico , Animais , Anticorpos Antiprotozoários , Formação de Anticorpos , Humanos , Coelhos , Proteínas Recombinantes , Estudos Soroepidemiológicos , Trofozoítos , UbiquitinaRESUMO
BACKGROUND: Amoebiasis is a parasitic disease caused by Entamoeba histolytica, which affects people living in low- and middle-income countries and has intestinal and extraintestinal manifestations. To date, knowledge on coronavirus disease 2019 (COVID-19) coinfection with enteric parasites is limited, and E. histolytica coinfection has not been previously described. Here we present the case of a patient with COVID-19 who, during hospitalisation, presented a clinical picture consistent with an amoebic liver abscess (ALA). CASE PRESENTATION: A 54-year-old man, admitted as a suspected case of COVID-19, presented to our hospital with dyspnoea, malaise, fever and hypoxaemia. A nasopharyngeal swab was positive for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) by reverse-transcription polymerase chain reaction. After 7 days, he developed diarrhoea, choluria and dysentery. An abdominal ultrasound showed a lesion compatible with a liver abscess; stool examination revealed E. histolytica trophozoites, and additional serology for E. histolytica was positive. After 12 days of treatment with metronidazole, ceftazidime and nitazoxanide, the patient reported acute abdominal pain, and an ultrasound examination revealed free liquid in the abdominal cavity. An emergency exploratory laparotomy was performed, finding 3000 mL of a thick fluid described as "anchovy paste". Computed tomography scan revealed a second abscess. He ended up receiving 21 days of antibiotic treatment and was discharged with satisfactory improvement. CONCLUSION: Here we present, to the best of our knowledge, the first report of ALA and COVID-19 co-presenting. Based on their pathophysiological similarities, coinfection with SARS-CoV-2 and E. histolytica could change the patient's clinical course; however, larger studies are needed to fully understand the interaction between these pathogens.
Assuntos
COVID-19 , Entamoeba histolytica , Abscesso Hepático Amebiano , Humanos , Abscesso Hepático Amebiano/diagnóstico , Abscesso Hepático Amebiano/tratamento farmacológico , Masculino , Metronidazol/uso terapêutico , Pessoa de Meia-Idade , SARS-CoV-2RESUMO
Amoebiasis is a protozoan disease caused by Entamoeba histolytica, and presents a geographic distribution of worldwide amplitude, high incidence, sometimes accompanied by severe clinical manifestations such as amoebic colitis and Amoebic Liver Abscess (ALA), remaining as a public health problem in developing countries. Entamoeba dispar is another species of amoeba that infects approximately 12% of the world's population, and it has previously been classified as noninvasive. However, E. dispar has already been isolated from patients with symptomatic non-dysenteric colitis, as well as its DNA sequences were detected and genotyped in samples from patients with dysenteric colitis, and patients with ALA, suggesting that this species could also be involved in the development of lesions in the large intestine and liver of human beings. In this context, this study aims to evaluate the ability of isolated strains of Entamoeba dispar in South America to cause liver damage, and to better characterize histopathological findings in 3, 8, 12 and 16 days after infection (DAI). Firstly, we assessed whether trophozoites from MCR, ACFN, ICS, ADO and VEJ E. dispar strains, and EGG Entamoeba histolytica strain differed in their in vitro phagocytosis ability, being related to greater ability to phagocyte with greater virulence. Then, we investigate and characterize histopathological changes present in the liver of mice induced by different strains of E. dispar. Our results demonstrated that trophozoites from E. dispar strains are capable of phagocyting human erythrocytes, but in lower amounts than Entamoeba histolytica. In addition, we described and characterized the lesions in different periods after infection by different E. dispar strains, and identified ACFN as the most pathogenic strain, followed by MCR. The large areas of necrosis produced by the ACFN strain as the eighth DAI, which also show high parasitism, led to 100% mortality. On the other hand, the ICS, ADO and VEJ strains did not produce mortality, and this was correlated with the presence of well-developed chronic granulomatous inflammation, necrosis absorption throughout the infection, and regeneration of the liver parenchyma. The greater pathogenicity of the ACFN strain strongly suggests that this strain could be producing higher levels of virulence factors. As the experimental infection, the heterogeneity of biological behavior of different Entamoeba dispar strains could be involved in the development of undiagnosed human clinical conditions.
Assuntos
Amoeba , Entamoeba histolytica , Entamoeba , Entamebíase , Abscesso Hepático Amebiano , Animais , Entamoeba/genética , Entamoeba histolytica/genética , Entamebíase/epidemiologia , Humanos , Cinética , Camundongos , VirulênciaRESUMO
Infection with the enteric protozoan Entamoeba histolytica is still a serious public health problem, especially in developing countries. Amoebic liver abscess (ALA) is the most common extraintestinal manifestation of the amoebiasis, and it can lead to serious and potentially life-threatening complications in some people. ALA can be cured by metronidazole (MTZ); however, because it has poor activity against luminal trophozoites, 40-60% of treated patients get repeated episodes of invasive disease and require repeated treatments that can induce resistance to MTZ, this may emerge as an important public health problem. Anti-virulence strategies that impair the virulence of pathogens are one of the novel approaches to solving the problem. In this study, we found that low doses of curcumin (10 and 50 µM) attenuate the virulence of E. histolytica without affecting trophozoites growth or triggering liver injury. Curcumin (CUR) decreases the expression of genes associated with E. histolytica virulence (gal/galnac lectin, ehcp1, ehcp5, and amoebapore), and is correlated with significantly lower amoebic invasion. In addition, oxidative stress is critically involved in the etiopathology of amoebic liver abscess; our results show no changes in mRNA expression levels of superoxide dismutase (SOD) and catalase (CAT) after E. histolytica infection, with or without CUR. This study provides clear evidence that curcumin could be an anti-virulence agent against E. histolytica, and makes it an attractive potential starting point for effective treatments that reduce downstream amoebic liver abscess.
RESUMO
Resumen El propósito de esta revisión es resaltar la importancia de la investigación en el área de la inmunología y su aplicación en el ámbito clínico. En una primera parte se presentan los descubrimientos más importantes que ayudaron a dilucidar los principales procesos fisiológicos involucrados en las enfermedades y de esta manera ayudaron a redireccionar la investigación en el área de la inmunología. Seguido, se describe un ejemplo de investigación básica relacionada con el papel de las citocinas en el absceso hepático amebiano, mostrando el trabajo de varios grupos de investigación en el mundo, con el objetivo de entender la respuesta inmune contra el parásito. Lo anterior nos permite argumentar la relevancia que tiene la investigación inmunológica dentro del contexto clínico.
Abstract The purpose of this review is to highlight the importance of research in immunology and its application in the clinical setting. The first part presents the most important discoveries that helped to elucidate the main physiological processes involved in the diseases and in this way helped to redirect research in immunology. Then, we describe an example of basic research related to the role of cytokines in the amoebic liver abscess, showing the work of several research groups in the world, with the aim of understanding the immune response against the parasite. This allows us to argue the relevance of immunological research within the clinical context.
Assuntos
Citocinas , Parasitos , Imunidade , Abscesso Hepático AmebianoRESUMO
Invasive amoebiasis is a life-threatening infection requiring immediate detection and treatment. However, diagnosis is challenging because conventional methods such as light microscopy and serology are unreliable. Molecular techniques are therefore considered the new diagnostic reference standard, but most of the developed assays are research tools and not widely available. Recently commercial multiplex PCR panels have been introduced which permit the simultaneous detection of multiple enteric pathogens including Entamoeba histolytica in stool samples. Our report demonstrates for the first time that these new assays might also serve as a rapid tool to diagnose amoebic liver abscess in patients with cystic focal liver lesions.
RESUMO
El óxido nítrico participa en funciones fisiológicas y fisiopatológicas, así como en el mecanismo de defensa del sistema inmunológico de mamíferos contra parásitos, virus y bacterias. La Entamoeba histolytica es un parásito protozoario causante de la amebiasis, la cual se caracteriza por el daño intestinal y la formación del absceso hepático amebiano (AHA). El desarrollo del absceso hepático amebiano en el hámster es similar al que desarrolla el humano, mientras que el ratón es resistente a la formación de este absceso, debido a un incremento en la producción de óxido nítrico. A diferencia del ratón, el desarrollo del absceso hepático amebiano en el hámster es debido a un exceso en la producción de óxido nítrico o posiblemente a una mayor susceptibilidad del hámster al daño producido por el óxido nítrico. Por lo tanto, sería importante realizar más estudios para determinar si en el humano, un exceso en la producción de óxido nítrico favorece la formación del absceso hepático amebiano.
Nitric oxide participates in both physiological and pathophysiological functions, and it plays an important role in the mammalian immune system in killing or inhibiting the growth of many pathogens, including parasites, viruses and bacteria. Entamoeba histolytica is a protozoan parasite that causes amoebiasis, which is characterized by intestinal damage and amoebic liver abscess development. The development of amoebic liver abscess in hamsters is similar to that in humans, whereas mice are resistant to amoebic liver abscess development due to an increase in nitric oxide production. Unlike in mice, amoebic liver abscess development in hamsters is due to an excess in nitric oxide production or possibly to a greater susceptibility of the hamster to damage caused by nitric oxide. Therefore, it could be important to elucidate if, in humans, an excess in nitric oxide production favors amoebic liver abscess development.