RESUMO
INTRODUCTION: Encephalitozoon cuniculi (E. cuniculi), a fungus that acts as an intracellular pathogen, causes a marked neurological syndrome in many host species and is a zoonotic concern. Although no well-established treatment for this syndrome is known, previous successful clinical experience using homeopathic phosphorus has been described in which symptom remission with no mortality occurred in 40/42 animals by means of unknown immunological mechanisms. The latter observation was the main motivation for this study. OBJECTIVE: To verify, in an in-vitro model, if macrophages infected with E. cuniculi can change in function after treatment with different potencies of phosphorus. MATERIALS AND METHODS: RAW 264.7 macrophages were infected with E. cuniculi in-vitro and treated with various homeopathic potencies of phosphorus. The vehicle was used as a control solution (0.06% succussed ethanol). After 1 and 24 hours, the following parameters were analyzed: parasite internalization (by the Calcofluor staining method), lysosome activity (by the acridine orange method), cytokine/chemokine production (by the MAGPIX system), and cell ultrastructure. Automatic image analysis was used when applicable, and the experiments were performed in triplicate. RESULTS: Treatment with vehicle alone increased interleukin (IL)-6, tumor necrosis factor alpha and monocyte chemotactic protein -1 production (p ≤ 0.05) and reduced the number of internalized parasites (p ≤ 0.001). A progressive and time-dependent increase in RANTES (regulated on activation, normal T-cell expressed and secreted) and lysosome activity (p ≤ 0.002) was observed only after treatment with the highest potency of phosphorus (Phos 200cH), together with decreased apoptosis rate, intense parasite digestion, and the presence of non-internalized spores. CONCLUSIONS: Phos 200 cH has a modulatory action on the activity of infected macrophages, especially a specific increase in RANTES, a key element in the prognosis of E. cuniculi-infected and of immunosuppressed patients bearing infections.
Assuntos
Encephalitozoon cuniculi/efeitos dos fármacos , Macrófagos/efeitos dos fármacos , Fósforo/uso terapêutico , Animais , Encephalitozoon cuniculi/patogenicidade , Encefalitozoonose/tratamento farmacológico , Homeopatia/métodos , Homeopatia/normas , Macrófagos/microbiologia , Fosfatos/uso terapêutico , CoelhosRESUMO
Microsporidia comprise a large group of obligate intracellular parasites. The microsporidian Encephalitozoon cuniculi causes disseminated infection in immunosuppressed patients with HIV, cancer, or transplants and in the elderly. In vivo and in vitro studies on the effectiveness of drugs are controversial. Currently, there is no effective treatment. We tested albendazole, albendazole sulfoxide, metronidazole, and cyclosporine in mice immunosuppressed with cyclophosphamide and inoculated by the intraperitoneal route with 10(7) E. cuniculi spores. One week after experimental inoculation, the mice were treated with albendazole, albendazole sulfoxide, metronidazole, and cyclosporine. Histological and morphometric analyses were performed to compare the treated groups. The state of immunosuppression was evaluated by phenotyping CD4(+) and CD8(+) T cells by flow cytometry. Nontreated mice showed acute disseminated and fatal encephalitozoonosis. The treatment with benzimidazoles significantly reduced infection until 30 days posttreatment (p.t.), but at 60 days p.t., the infection had recurred. Metronidazole decreased infection by a short time, and cyclosporine was not effective. All animals were immunosuppressed by all the experiments, as demonstrated by the low number of CD4(+) and CD8(+) T cells. We conclude that no drug was effective against E. cuniculi, but the benzimidazoles controlled the infection transiently.
Assuntos
Albendazol/análogos & derivados , Albendazol/uso terapêutico , Ciclosporina/uso terapêutico , Encefalitozoonose/tratamento farmacológico , Metronidazol/uso terapêutico , Albendazol/farmacologia , Animais , Linfócitos T CD4-Positivos/efeitos dos fármacos , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD8-Positivos/efeitos dos fármacos , Linfócitos T CD8-Positivos/imunologia , Ciclofosfamida/farmacologia , Ciclofosfamida/uso terapêutico , Ciclosporina/farmacologia , Encephalitozoon cuniculi/efeitos dos fármacos , Hospedeiro Imunocomprometido , Intestinos/microbiologia , Intestinos/patologia , Rim/microbiologia , Rim/patologia , Fígado/microbiologia , Fígado/patologia , Pulmão/microbiologia , Pulmão/patologia , Masculino , Metronidazol/farmacologia , Camundongos , Camundongos Endogâmicos BALB C , Baço/microbiologia , Baço/patologiaRESUMO
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.
Assuntos
Síndrome da Imunodeficiência Adquirida/complicações , Doenças do Gato/microbiologia , Encephalitozoon/classificação , Encefalitozoonose/veterinária , Albendazol/uso terapêutico , Animais , Anti-Helmínticos/uso terapêutico , Gatos , Encefalitozoonose/complicações , Encefalitozoonose/tratamento farmacológico , Encefalitozoonose/microbiologia , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
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.
Assuntos
Ciclosporina , Encephalitozoon , Encefalitozoonose/tratamento farmacológico , Imunossupressores , Animais , Ciclosporina/metabolismo , Ciclosporina/farmacologia , Ciclosporina/uso terapêutico , Encephalitozoon/efeitos dos fármacos , Encephalitozoon/imunologia , Fezes/microbiologia , Humanos , Imunoglobulina G/metabolismo , Imunossupressores/metabolismo , Imunossupressores/farmacologia , Imunossupressores/uso terapêutico , Intestinos/microbiologia , Intestinos/patologia , Camundongos , Camundongos Endogâmicos C57BL , Distribuição AleatóriaRESUMO
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.