RESUMEN
The microsporidia are a large group of intracellular parasites with a broad range of hosts, including humans. Encephalitozoon intestinalis is the second microsporidia species most frequently associated with gastrointestinal disease in humans, especially immunocompromised or immunosuppressed individuals, including children and the elderly. The prevalence reported worldwide in these groups ranges from 0 to 60%. Currently, albendazole is most commonly used to treat microsporidiosis caused by Encephalitozoon species. However, the results of treatment are variable, and relapse can occur. Consequently, efforts are being directed toward identifying more effective drugs for treating microsporidiosis, and the study of new molecular targets appears promising. These parasites lack mitochondria, and oxidative phosphorylation therefore does not occur, which suggests the enzymes involved in glycolysis as potential drug targets. Here, we have for the first time characterized the glycolytic enzyme triosephosphate isomerase of E. intestinalis at the functional and structural levels. Our results demonstrate the mechanisms of inactivation of this enzyme by thiol-reactive compounds. The most striking result of this study is the demonstration that established safe drugs such as omeprazole, rabeprazole and sulbutiamine can effectively inactivate this microsporidial enzyme and might be considered as potential drugs for treating this important disease.
Asunto(s)
Albendazol/uso terapéutico , Proteínas Fúngicas/antagonistas & inhibidores , Microsporidios/efectos de los fármacos , Microsporidiosis/tratamiento farmacológico , Triosa-Fosfato Isomerasa/antagonistas & inhibidores , Secuencia de Aminoácidos , Encephalitozoon/efectos de los fármacos , Encephalitozoon/enzimología , Encephalitozoon/genética , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Enfermedades Gastrointestinales/tratamiento farmacológico , Enfermedades Gastrointestinales/microbiología , Regulación Enzimológica de la Expresión Génica/efectos de los fármacos , Regulación Fúngica de la Expresión Génica/efectos de los fármacos , Humanos , Microsporidios/enzimología , Microsporidios/genética , Microsporidiosis/microbiología , Omeprazol/uso terapéutico , Rabeprazol/uso terapéutico , Homología de Secuencia de Aminoácido , Tiamina/análogos & derivados , Tiamina/uso terapéutico , Triosa-Fosfato Isomerasa/genética , Triosa-Fosfato Isomerasa/metabolismoRESUMEN
Microsporidia are a group of intracellular pathogens causing self-limited and severe diseases in immunocompetent and immunocompromised individuals, respectively. A cellular type 1 adaptive response, mediated by IL-12, IFNγ, CD4+, and CD8+ T cells has been shown to be essential for host resistance, and dendritic cells (DC) play a key role at eliciting anti-microsporidial immunity. We investigated the in vitro response of DC and DC precursors/progenitors to infection with Encephalitozoon intestinalis (Ei), a common agent of human microsporidosis. Ei-exposed DC cultures up-regulated the surface expression of MHC class II and the costimulatory molecules CD86 and CD40, only when high loads of spores were used. A vigorous secretion of IL-6 but not of IL-1ß or IL-12p70 was also observed in these cultures. Ei-exposed DC cultures consisted of immature infected and mature bystander DC, as assessed by MHC class II and costimulatory molecules expression, suggesting that intracellular Ei spores deliver inhibitory signals in DC. Moreover, Ei selectively inhibited the secretion of IL-12p70 in LPS-stimulated DC. Whereas Ei-exposed DC promoted allogeneic naïve T cell proliferation and IL-2 and IFNγ secretion in DC-CD4+ T cell co-cultures, separated co-cultures with bystander or infected DCs showed stimulation or inhibition of IFNγ secretion, respectively. When DC precursors/progenitors were exposed to Ei spores, a significant inhibition of DC differentiation was observed without shifting the development toward cells phenotypically or functionally compatible with myeloid-derived suppressor cells. Neutralization experiments demonstrated that this inhibitory effect is IL-6-dependent. Altogether this investigation reveals a novel potential mechanism of immune escape of microsporidian parasites through the modulation of DC differentiation and maturation.
Asunto(s)
Células Dendríticas/citología , Células Dendríticas/inmunología , Encephalitozoon/inmunología , Encefalitozoonosis/inmunología , Evasión Inmune/inmunología , Interleucina-6/inmunología , Animales , Antígeno B7-2/biosíntesis , Linfocitos T CD4-Positivos/inmunología , Antígenos CD40/biosíntesis , Linfocitos T CD8-positivos/inmunología , Diferenciación Celular/inmunología , Células Cultivadas , Encefalitozoonosis/microbiología , Interferón gamma/inmunología , Interferón gamma/metabolismo , Subunidad p35 de la Interleucina-12/inmunología , Activación de Linfocitos/inmunología , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Esporas Bacterianas/inmunologíaRESUMEN
Microsporidioses are considered emerging and opportunistic infections in immunocompromised individuals worldwide. The purpose of this study was to identify the species of intestinal microsporidia in patients with HIV-AIDS from the Servicio Autónomo Hospital Universitario de Maracaibo, Venezuela (SAHUM). Fecal samples were collected from 50 patients with confirmed diagnosis of HIV, during the years 2007 and 2008; the CD4 values were obtained from 42 patients. The samples were analyzed by separate PCRs to identify Encephalitozoon intestinalis and Enterocytozoon bieneusi. Microsporidia species showed a 36% prevalence: ten patients had Encephalitozoon intestinalis, four Enterocytozoon bieneusi and four both species. An inverse and statistically significant relationship between the CD4 count and the presence of microsporidia in the fecal sample was also found. It is remarkable the high prevalence of microsporidia species observed in the HIV patients studied, with a predominance of E. intestinalis.
Asunto(s)
Diarrea/epidemiología , Encephalitozoon/aislamiento & purificación , Encefalitozoonosis/epidemiología , Enterocytozoon/aislamiento & purificación , Heces/microbiología , Infecciones por VIH/epidemiología , Microsporidiosis/epidemiología , Infecciones Oportunistas Relacionadas con el SIDA/epidemiología , Infecciones Oportunistas Relacionadas con el SIDA/inmunología , Infecciones Oportunistas Relacionadas con el SIDA/microbiología , Adulto , Enfermedades Asintomáticas , Recuento de Linfocito CD4 , Coinfección , Comorbilidad , ADN de Hongos/análisis , Diarrea/microbiología , Encefalitozoonosis/microbiología , Femenino , Infecciones por VIH/inmunología , Síndrome de Emaciación por VIH/epidemiología , Humanos , Huésped Inmunocomprometido , Masculino , Microsporidiosis/microbiología , Persona de Mediana Edad , Prevalencia , Venezuela/epidemiología , Adulto JovenRESUMEN
Los microsporidios pueden provocar infecciones emergentes y oportunistas en individuos inmunocomprometidos de todo el mundo. Se realizó éste estudio para identificar las especies de microsporidios intestinales presentes en pacientes con VIH-SIDA del Servicio Autónomo Hospital Universitario de Maracaibo (SAHUM). Se recolectaron 50 muestras fecales de individuos con diagnóstico confirmado de VIH durante los años 2007-2008; se obtuvieron las cifras de CD4 de solo 42 pacientes. Las muestras se analizaron mediante PCR separadas para la identificación de Encephalitozoon intestinalis y Enterocytozoon bieneusi. Las especies de microsporidios presentaron un 36% de prevalencia, 10 pacientes presentaron Encephalitozoon intestinalis, 4 Enterocytozoon bieneusi y 4 ambas especies. Se determinó una relación inversamente proporcional y estadísticamente significativa entre el contaje de CD4 y la presencia de microsporidios en la muestra fecal. Es destacable la elevada prevalencia de especies de microsporidios observada en los pacientes VIH estudiados, donde predominó E. intestinalis.
Microsporidioses are considered emerging and opportunistic infections in immunocompromised individuals worldwide. The purpose of this study was to identify the species of intestinal microsporidia in patients with HIV-AIDS from the Servicio Autónomo Hospital Universitario de Maracaibo, Venezuela (SAHUM). Fecal samples were collected from 50 patients with confirmed diagnosis of HIV, during the years 2007 and 2008; the CD4 values were obtained from 42 patients. The samples were analyzed by separate PCRs to identify Encephalitozoon intestinalis and Enterocytozoon bieneusi. Microsporidia species showed a 36% prevalence: ten patients had Encephalitozoon intestinalis, four Enterocytozoon bieneusi and four both species. An inverse and statistically significant relationship between the CD4 count and the presence of microsporidia in the fecal sample was also found. It is remarkable the high prevalence of microsporidia species observed in the HIV patients studied, with a predominance of E. intestinalis.
Asunto(s)
Adulto , Femenino , Humanos , Masculino , Persona de Mediana Edad , Adulto Joven , Diarrea/epidemiología , Encephalitozoon/aislamiento & purificación , Encefalitozoonosis/epidemiología , Enterocytozoon/aislamiento & purificación , Heces/microbiología , Infecciones por VIH/epidemiología , Microsporidiosis/epidemiología , Infecciones Oportunistas Relacionadas con el SIDA/epidemiología , Infecciones Oportunistas Relacionadas con el SIDA/inmunología , Infecciones Oportunistas Relacionadas con el SIDA/microbiología , Enfermedades Asintomáticas , Coinfección , Comorbilidad , ADN de Hongos/análisis , Diarrea/microbiología , Encefalitozoonosis/microbiología , Infecciones por VIH/inmunología , Síndrome de Emaciación por VIH/epidemiología , Huésped Inmunocomprometido , Microsporidiosis/microbiología , Prevalencia , Venezuela/epidemiologíaRESUMEN
BACKGROUND: Microsporidiosis is a life threatening opportunistic infection of AIDS patients. The infection is usually restricted to specific anatomical areas, but could become systemic depending on the involved species. Genital microsporidiosis in female patients is rare. OBJECTIVE: To report genital microsporidiosis in female AIDS patients. METHODS: Tissues samples from the genital tract (ovary, fallopian tubes and uterus) of eight deceased women who died of wasting syndrome associated to AIDS and disseminated microsporidiosis at the Institute of Tropical Medicine Pedro Kourí were collected between 1997 and 2005. Using an indirect immunohistochemistry assay the microsporidia species involved in those cases were identified. RESULTS: We report several cases of microsporidial infection of the female genital tract. Six out of eight women with the disseminated form of the disease showed the presence of microsporidia in the genital tract. Encephalitozoon cuniculi and Encephalitozoon hellem were identified in the internal lining epithelium of the fallopian tubes and endometrium. CONCLUSIONS: Microsporidia species could disseminate to other organs and become systemic in severe immunocompromised cases. To our knowledge this is the greatest number of female genital tract microsporidiosis cases so far reported in humans.
Asunto(s)
Infecciones Oportunistas Relacionadas con el SIDA/patología , Encefalitozoonosis/patología , Enfermedades de los Genitales Femeninos/patología , Infecciones Oportunistas Relacionadas con el SIDA/microbiología , Autopsia , Vasos Sanguíneos/microbiología , Cuello del Útero/microbiología , Progresión de la Enfermedad , Encephalitozoon/aislamiento & purificación , Encephalitozoon cuniculi/aislamiento & purificación , Encefalitozoonosis/microbiología , Endometrio/microbiología , Células Epiteliales/microbiología , Trompas Uterinas/microbiología , Femenino , Enfermedades de los Genitales Femeninos/microbiología , Síndrome de Emaciación por VIH/patología , Humanos , Macrófagos/microbiología , Especificidad de Órganos , Esporas Fúngicas/aislamiento & purificaciónRESUMEN
Microsporidia are eukaryotic, intracellular obligate parasites that infect invertebrate and vertebrate animals, and have emerged as important opportunistic parasites in AIDS patients. We used light microscopy to detect microsporidial spores in stool samples of a domestic cat confirmed as Encephalitozoon intestinalis by PCR, owned by an AIDS patient with chronic diarrhea and E. intestinalis infection. Cats can be considered hosts of E. intestinalis.
Asunto(s)
Síndrome de Inmunodeficiencia Adquirida/complicaciones , Enfermedades de los Gatos/microbiología , Encephalitozoon/clasificación , Encefalitozoonosis/veterinaria , Albendazol/uso terapéutico , Animales , Antihelmínticos/uso terapéutico , Gatos , Encefalitozoonosis/complicaciones , Encefalitozoonosis/tratamiento farmacológico , Encefalitozoonosis/microbiología , Humanos , Masculino , Persona de Mediana EdadRESUMEN
INTRODUCTION: Microsporidia are intracellular micro-organisms, characterized by mature spores with chitin walls and by one extrusive polar tube through which they pour their sporoplasm to the host cells. In immunocompromised patients, Enterocytozoon bieneusi and Encephalitozoon intestinalis produce diarrhea and systemic dissemination. In Mexico there is not information about microsporidia in children with cancer. OBJECTIVE: The aim of this pilot study was to investigate the presence of microsporidia species in pediatric patients with leukemia or lymphoma. MATERIAL AND METHODS: We obtained fecal samples from thirteen patients. The samples were processed to detect microsporidia by both modified Ziehl-Neelsen and clacofluor white stains, DNA was isolated to amplify rRNA specific sequences, to identify E. bieneusi, E. intestinalis, E. cuniculi and E. hellem by DNA polymerase chain reaction (PCR). Other parasites and pathogenic bacteria were also tested. RESULTS: Based on morphologic traits 7/13 samples were found positives to microsporidia and 6/10 by PCR. Was identified E. bieneusi in three patients with leukemia and one with lymphoma, another two children with leukemia were infected with E. intestinalis. Almost all children were high-risk patients and in phase of re-induction, consolidation or with many chemotherapy treatments. All the patients with microspiridia did not present diarrhea at the moment of the sampling; however, in two children with diarrhea it was found Cyclospora cayetanensis. Also we obtained feces from five patients' mothers and microsporidia spores were identified by stain in all of them and by PCR it was diagnosed the species in three of them. CONCLUSION: It was demonstrated that the feces of patients with leukemia or lymphoma had microsporidia, therefore is necessary to know the prevalence of these microorganisms and to analyze their impact in evolution of cancer patients.
Asunto(s)
Leucemia/epidemiología , Linfoma no Hodgkin/epidemiología , Microsporidiosis/epidemiología , Adolescente , Adulto , Niño , Preescolar , Comorbilidad , Cyclospora/aislamiento & purificación , Ciclosporiasis/epidemiología , Ciclosporiasis/parasitología , Encephalitozoon/aislamiento & purificación , Encefalitozoonosis/epidemiología , Enterocytozoon/aislamiento & purificación , Heces/microbiología , Heces/parasitología , Femenino , Humanos , Lactante , Masculino , México/epidemiología , Microsporidiosis/microbiología , Madres , Proyectos Piloto , Leucemia-Linfoma Linfoblástico de Células Precursoras B/epidemiologíaRESUMEN
Relata-se um caso de ceratoconjuntivite causada por Encephalitozoon hellem em agapornis (Agapornis spp.) adultos, provenientes de um criatório comercial. Cinco animais apresentaram sinais clínicos de ceratoconjuntivite, blefaroespasmo e blefaroedema bilateral, com presença de secreção seropurulenta. Amostras fecais foram colhidas e foi realizado exame coproparasitológico, com resultado negativo. Dois animais foram necropsiados, sendo detectados, em impressões de raspado de conjuntiva ocular, esporos e outros estádios evolutivos de Microsporidium. A confirmação do diagnóstico foi feita pela reação em cadeia de polimerase e sequenciamento de fragmentos amplificados, com utilização de primers específicos para o gene da subunidade 18S do rRNA de E. hellem. A análise dos fragmentos amplificados demonstrou 100 por cento de similaridade com outras sequências de E. hellem publicadas no GenBank. Este é primeiro relato de infecção por E. hellem em aves no Brasil.(AU)
A clinical case of keratoconjunctivitis by Encephalitozoon hellem in adult lovebirds (Agapornis spp.) from a commercial flock is reported. Five animals presented clinical symptoms of keratoconjunctivitis, blepharospasm, and bilateral blepharoedema, with seropurulent secretion. Coproparasitological diagnosis was carried out in fecal samples, with negative results. Two animals were necropsied, with detection of spores and other developmental stages of Microsporidium in conjunctival smears. The confirmation of the diagnosis was accomplished by the polimerase chain reaction with specific primers for 18S subunit of the rRNA of E. hellem, followed by sequencing of amplified fragments, which revealed 100 percent of genetic similarity to E. hellem. This study is the first report of E. hellem infection in birds in Brazil.(AU)
Asunto(s)
Animales , Queratoconjuntivitis/diagnóstico , Queratoconjuntivitis/veterinaria , Encephalitozoon/aislamiento & purificación , Periquitos , ZoonosisRESUMEN
Relata-se um caso de ceratoconjuntivite causada por Encephalitozoon hellem em agapornis (Agapornis spp.) adultos, provenientes de um criatório comercial. Cinco animais apresentaram sinais clínicos de ceratoconjuntivite, blefaroespasmo e blefaroedema bilateral, com presença de secreção seropurulenta. Amostras fecais foram colhidas e foi realizado exame coproparasitológico, com resultado negativo. Dois animais foram necropsiados, sendo detectados, em impressões de raspado de conjuntiva ocular, esporos e outros estádios evolutivos de Microsporidium. A confirmação do diagnóstico foi feita pela reação em cadeia de polimerase e sequenciamento de fragmentos amplificados, com utilização de primers específicos para o gene da subunidade 18S do rRNA de E. hellem. A análise dos fragmentos amplificados demonstrou 100 por cento de similaridade com outras sequências de E. hellem publicadas no GenBank. Este é primeiro relato de infecção por E. hellem em aves no Brasil.
A clinical case of keratoconjunctivitis by Encephalitozoon hellem in adult lovebirds (Agapornis spp.) from a commercial flock is reported. Five animals presented clinical symptoms of keratoconjunctivitis, blepharospasm, and bilateral blepharoedema, with seropurulent secretion. Coproparasitological diagnosis was carried out in fecal samples, with negative results. Two animals were necropsied, with detection of spores and other developmental stages of Microsporidium in conjunctival smears. The confirmation of the diagnosis was accomplished by the polimerase chain reaction with specific primers for 18S subunit of the rRNA of E. hellem, followed by sequencing of amplified fragments, which revealed 100 percent of genetic similarity to E. hellem. This study is the first report of E. hellem infection in birds in Brazil.
Asunto(s)
Animales , Queratoconjuntivitis/diagnóstico , Queratoconjuntivitis/veterinaria , Encephalitozoon/aislamiento & purificación , Periquitos , ZoonosisRESUMEN
We describe a new microsporidian species, Encephalitozoon romaleae n. sp., isolated from an invertebrate host, the grasshopper Romalea microptera, collected near Weeks Island, Louisiana, and Jacksonville, Florida. This microsporidian is characterized by specificity to the gastric caecae and midgut tissues of the host and a life cycle that is nearly identical to that of Encephalitozoon hellem and Encephalitozoon cuniculi. Mature spores are larger (3.97 x 1.95 microm) than those of other Encephalitozoon species. Polar filament coils number 7 to 8 in a single row. Analysis of the small subunit (SSU) rDNA shows that E. romaleae fits well into the Encephalitozoon group and is a sister taxon to E. hellem. This is the first Encephalitozoon species that has been shown to complete its life cycle in an invertebrate host.
Asunto(s)
Encephalitozoon/clasificación , Saltamontes/parasitología , Animales , ADN de Hongos/química , ADN Ribosómico/química , Encephalitozoon/genética , Encephalitozoon/ultraestructura , Microscopía Electrónica de Transmisión , Microscopía de Contraste de Fase , FilogeniaRESUMEN
INTRODUCTION: Microsporidia are obligate intracellular parasites that are recognized as important opportunistic pathogens of immunocompromised and transplanted patients. Enterocytozoon bieneusi and, less frequently, Encephalitozoon intestinalis are the most prevalent species in humans; both of them are associated with enteric infections. Cell cultures have been useful in the study of microsporidia biology. In Colombia, however, no isolates of microsporidia from patients with AIDS have been obtained. OBJECTIVE: A cell culture of intestinal microsporidia was established from stools of positive patients in order to isolate a native strain. MATERIALS AND METHODS: Stool from a single AIDS patient was concentrated with the water-ether technique, and the sediment was treated with a mixture of antibiotics and antifungal agents for 18 hours at 37 degrees C. Vero cells were cultivated in 24-well plates with Gibco RPMI medium supplemented with 10% bovine fetal serum and antibiotics. The culture was subsequently inoculated with previously concentrated spores. The medium was changed every second day and the presence of spores was evaluated with the Quick Hot Gram chromotrope stain. RESULTS: Two weeks post-infection, microsporidial spores were identified with characteristic morphology and staining properties. PCR results showed that Encephalitozoon intestinalis was the isolated species. CONCLUSIONS: A cell culture of microsporidia was established from a stool sample. This protocol is important to isolate and maintain additional native Colombian strains and it will contribute to biochemical, immunological and epidemiological studies of the currently established strain.
Asunto(s)
Síndrome de Inmunodeficiencia Adquirida/parasitología , Encephalitozoon/aislamiento & purificación , Heces/parasitología , Microsporidios/aislamiento & purificación , Animales , Bovinos , Línea Celular , Colombia , Humanos , Persona de Mediana EdadRESUMEN
Introducción. Los microsporidios son agentes de infecciones oportunistas en pacientes con sida y con trasplantes, principalmente. Enterocytozoon bieneusi y Encephalitozoon intestinalis son los más frecuentes, asociados con infecciones entéricas. Los cultivos celulares han contribuido al conocimiento de los microsporidios. En Colombia no se han obtenido aislamientos provenientes de pacientes con microsporidiosis y, por consiguiente, no existen cepas autóctonas de los mismos. Objetivo. Establecer el cultivo celular de microsporidios intestinales a partir de materia fecal de pacientes parasitados. Materiales y métodos. Se realizó concentración agua-éter de la materia fecal positiva para microsporidios y el sedimento resultante se trató con una mezcla de antibióticos y antimicóticos durante 18 horas a 37 oC. Se inocularon células Vero previamente cultivadas en placas de 24 pozos y en medio RPMI con suplemento de suero bovino fetal al 10 por ciento y antibióticos, con las esporas concentradas. Los cultivos se mantuvieron a 37 oC al 5 por ciento de CO2. Se cambió de medio cada dos días y se evaluó la presencia de esporas en los sobrenadantes mediante Gram-cromótropo rápido en caliente. Resultados. En la segunda semana después de la infección, se encontraron esporas de microsporidios con morfología y coloración características. Mediante PCR se determinó que el microsporidio encontrado correspondía a la especie E. intestinalis. Conclusión. Se estableció el cultivo in vitro de microsporidios de materia fecal. Este protocolo es importante para la obtención y el mantenimiento de cepas autóctonas en Colombia, y contribuirá a las investigaciones de aspectos bioquímicos, inmunológicos y epidemiológicos de dichas cepas.
Asunto(s)
Síndrome de Inmunodeficiencia Adquirida , Encephalitozoon , Encephalitozoon/aislamiento & purificación , Técnicas In Vitro , Microsporidiosis , Células Cultivadas , HecesRESUMEN
The present paper reports the occurrence of natural co-infection of Babesia caballi and Encephalitozoon-like microsporidia in the tick Anocentor nitens. Engorged females of ticks, collected from a naturally B. caballi-infected horse, were incubated at 27 degrees C and relative humidity over 83%. After a 6-day incubation period, Giemsa-stained smears prepared from hemolymph were examined microscopically under oil immersion. B. caballi infected ticks were dissected and samples of midgut tissue were examined by transmission electron microscopy, through which free sporokinetes were seen in the cytoplasm of gut epithelial cells. In addition, Encephalitozoon-like microsporidia were observed inside the parasitophorous vacuoles in the same cell in which sporokinetes of B. caballi were found and also in some neighbour cells. They presented different morphological stages, suggesting a sequential phases of development.
Asunto(s)
Babesia/fisiología , Encephalitozoon/fisiología , Ixodidae/microbiología , Ixodidae/parasitología , Animales , Babesia/aislamiento & purificación , Babesia/ultraestructura , Babesiosis/parasitología , Babesiosis/transmisión , Babesiosis/veterinaria , Sistema Digestivo/microbiología , Sistema Digestivo/parasitología , Sistema Digestivo/ultraestructura , Encephalitozoon/aislamiento & purificación , Encephalitozoon/ultraestructura , Femenino , Enfermedades de los Caballos/parasitología , Enfermedades de los Caballos/transmisión , Caballos , Ixodidae/ultraestructura , Estadios del Ciclo de Vida , Microscopía Electrónica de TransmisiónRESUMEN
INTRODUCTION: Encephalitozoon intestinalis, a parasite belonging to the phylum Microsporidia, is causes gastrointestinal infections in the immunocompromised host. A suitable pharmacologically immunosuppressed animal model for the study of natural E. intestinalis infection, which can establish the immune components that respond to this parasite, is lacking. OBJECTIVE: To evaluate the effect of immunosuuppression with Cyclosporine A (CsA) in C57BL/ 6 mice on experimental infection with E. intestinalis infection. MATERIALS AND METHODS: Eighty C57BL/6 mice were distributed in four treatment groups: Control, CsA-immunosuppressed mice without infection, immunocompetent and immunossuppressed mice infected with E. intestinalis. Mice were immunosuppressed with a weekly dose of 50 mg/Kg body weight of CsA, during the course of the study. Five mice from each group were sacrificed 2, 3, 4 and 6 weeks post-infection, to obtain blood for antibody testing and stool samples were analyzed to assess excretion of spores. RESULTS: Production of specific IgG antibodies was significantly higher in the immunocompetent group as compared to the immunosuppressed group of experimentally infected mice. In the infected mice, parasites were not observed in any tissues different from the small intestine. However, spore excretion through the stool and duodenal liquid was higher in the group of immunosuppresed infected mice. CONCLUSION: Immunosuppression induced with CsA in the murine model did not allow parasite dissemination and illness progression, but raised kinetics of spore excretion and decreased the production of IgG antibodies.
Asunto(s)
Ciclosporina , Encephalitozoon , Encefalitozoonosis/tratamiento farmacológico , Inmunosupresores , Animales , Ciclosporina/metabolismo , Ciclosporina/farmacología , Ciclosporina/uso terapéutico , Encephalitozoon/efectos de los fármacos , Encephalitozoon/inmunología , Heces/microbiología , Humanos , Inmunoglobulina G/metabolismo , Inmunosupresores/metabolismo , Inmunosupresores/farmacología , Inmunosupresores/uso terapéutico , Intestinos/microbiología , Intestinos/patología , Ratones , Ratones Endogámicos C57BL , Distribución AleatoriaRESUMEN
Introducción. Encephalitozoon intestinalis es un microsporidio parásito del intestino, que puedediseminarse en pacientes inmunocomprometidos. Existen referencias de modelos animales inmunosuprimidos para el estudio de la microsporidiosis utilizando fármacos que producen supresión total de la respuesta inmune; sin embargo, no se han estudiado los efectos deinmunosupresores con acción selectiva sobre los componentes de esta respuesta. Objetivo. Evaluar el efecto de la inmunosupresión con ciclosporina A (CsA) en ratones C57BL/ 6 infectados con E. intestinalis.Materiales y métodos. Se utilizaron 80 ratones C57BL/6 distribuidos en cuatro grupos: infectados, inmunosuprimidos e infectados, inmunosuprimidos no infectados y controles. La inmunosupresión con CsA (50 mg/kg) se realizó vía intraperitoneal durante todo el estudio. En la semanas 2, 3, 4 y 6 posteriores a la infección se obtuvo sangre para determinar los anticuerpos, y materia fecal para evaluar la cinética de excreción de esporas. Además, se extrajeron varios órganos para estudiar la histopatología y observar la posible diseminación del parásito. Resultados. La producción de anticuerpos IgG fue mayor en los ratones inmunocompetentes infectados que en los inmunosuprimidos infectados con E. intestinalis. No se encontró elparásito en órganos diferentes al intestino delgado en los dos grupos infectados. Sin embargo, la excreción de esporas, tanto en heces como en líquido duodenal, fue mayor en el grupo inmunosuprimido infectado. Conclusión. La CsA en el modelo en ratón no indujo la diseminación de E. intestinalis ni la exacerbación de la enfermedad, pero contribuyó al aumento en la cinética de excreción deesporas y la disminución de la producción de anticuerpos IgG en los ratones inmunosuprimidos infectados.
Asunto(s)
Ratas , Ciclosporina , Encephalitozoon , Microsporidiosis , Microsporidia no Clasificados/inmunología , Formación de Anticuerpos , Terapia de InmunosupresiónRESUMEN
Spores of four species of microsporidia isolated from humans were analyzed by matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF MS) and specific biomarkers were found for each. The microsporidia analyzed included three species, Encephalitozoon cuniculi, Encephalitozoon hellem, and Encephalitozoon intestinalis and the fourth organism is the recently described Brachiola algerae. Whole spores, spore shells, and soluble fractions were applied directly to the MALDI target without further purification steps. MALDI-TOF MS analysis of both whole spores and soluble fractions of the four isolates revealed a group of unique, characteristic, and reproducible spectral markers in the mass range of 2,000-8,000 Da. Statistical analysis of the averaged centroided masses uncovered two distinct sets of unique peptides or biomarkers, one originated from whole spores and the other from soluble fractions, that can differentiate the four microsporidian species studied. MALDI-TOF MS analysis of whole organisms is a rapid, sensitive, and specific option to characterize microsporidian isolates and has the potential for several applications in parasitology.
Asunto(s)
Apansporoblastina/química , Encephalitozoon/química , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción , Animales , Apansporoblastina/clasificación , Apansporoblastina/aislamiento & purificación , Encephalitozoon/aislamiento & purificación , Encephalitozoon cuniculi/aislamiento & purificación , Encefalitozoonosis/diagnóstico , Encefalitozoonosis/epidemiología , Encefalitozoonosis/veterinaria , Humanos , Microsporidios/clasificación , Microsporidios/aislamiento & purificación , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción/métodos , Esporas Protozoarias/clasificación , Esporas Protozoarias/aislamiento & purificación , Coloración y EtiquetadoRESUMEN
Human enteropathogenic microsporidia (HEM), Cryptosporidium parvum, Cyclospora cayetanesis, and Giardia lamblia are associated with gastrointestinal disease in humans. To date, the mode of transmission and environmental occurrence of HEM (Encephalitozoon intestinalis and Enterocytozoon bieneusi) and Cyclospora cayetanesis have not been fully elucidated due to lack of sensitive and specific environmental screening methods. The present study was undertaken with recently developed methods, to screen various water sources used for public consumption in rural areas around the city of Guatemala. Water concentrates collected in these areas were subjected to community DNA extraction followed by PCR amplification, PCR sequencing and computer database homology comparison (CDHC). All water samples screened in this study had been previously confirmed positive for Giardia spp. by immunofluorescent assay (IFA). Of the 12 water concentrates screened, 6 showed amplification of microsporidial SSU-rDNA and were subsequently confirmed to be Encephalitozoon intestinalis. Five of the samples allowed for amplification of Cyclospora 18S-rDNA; three of these were confirmed to be Cyclospora cayetanesis while two could not be identified because of inadequate sequence information. Thus, this study represents the first confirmed identification of Cyclospora cayetanesis and Encephalitozoon intestinalis in source water used for consumption. The fact that the waters tested may be used for human consumption indicates that these emerging protozoa may be transmitted by ingestion of contaminated water.
Asunto(s)
Cryptosporidium parvum/aislamiento & purificación , Cyclospora/aislamiento & purificación , ADN Protozoario/análisis , Encephalitozoon/aislamiento & purificación , Abastecimiento de Agua/normas , Animales , Cryptosporidium parvum/genética , Cyclospora/genética , Recolección de Datos , Encephalitozoon/genética , Guatemala , Reacción en Cadena de la Polimerasa , Salud Pública , Población RuralRESUMEN
La microsporidiosis intestinal es la infección del tracto digestivo alto, Enterocytozoon bieneusi y Encephalolytozoon intestinalis, pertenecientes al phylum Microspora, que en el hospedero inmunocomprometido especialmente con el síndrome de inmunodeficiencia adquirida (SIDA), produce cuadros de diarrea prolongada y malabsorción. Se revisarán sus aspectos históricos, biológicos, fisiopatológicos, inmunológicos, epidemiológicos, clínicos y de tratamiento. Método: para la elaboración de esta revisión se emplearon las bases de datos Medline y Pubmed a partir de 1981 hasta la fecha
Asunto(s)
Técnicas de Laboratorio Clínico , Encephalitozoon , Enterocytozoon , Microsporidiosis , Infecciones Oportunistas Relacionadas con el SIDA , Guías como AsuntoRESUMEN
We present the case of a four year old boy with a history of repeated upper respiratory tract infections and pyoderma. He presented fever, seizures, inhability to talk, loss of swallowing, fine tremor in the upper extremities; positive bilateral Babinski reflex and quadriparesis. The diagnosis of Brutonïs disease and generalized microporidiosis was based on immunologic analysis, smear tests with chromotrope R2 stain and indirect immunofluorescense with monoclonal 3B6 antibody for Encephalitozoon species in samples of spinal fluid, bronchial and paranasal sinus aspirates and stool, which were all positive. The patient was treated with albendazol during 72 days; he left the hospital in a good condition, walking, talking and able to swallow. His laboratory test controls were negative; he is followed up in the outpatient department
Asunto(s)
Humanos , Masculino , Niño , Agammaglobulinemia , Encephalitozoon , Encefalitozoonosis , Animales Domésticos/parasitología , Quimioterapia Combinada , Encephalitozoon , Encefalitozoonosis , Eritromicina , México , Piodermia , SulfisoxazolRESUMEN
We present the case of a four-year-old boy with a history of repeated upper respiratory tract infections and pyoderma. He presented fever, seizures, inability to talk, loss of swallowing, fine tremor in the upper extremities; positive bilateral Babinski reflex and quadriparesis. The diagnosis of Bruton's disease and generalized microporidiosis was based on immunologic analysis, smear tests with chromotrope R2 stain and indirect immunofluorescense with monoclonal 3B6 antibody for Encephalitozoon species in samples of spinal fluid, bronchial and paranasal sinus aspirates and stool, which were all positive. The patient was treated with albendazol during 72 days; he left the hospital in a good condition, walking, talking and able to swallow. His laboratory test controls were negative; he is followed up in the outpatient department.