RESUMO
Coccidiosis, an intestinal disease caused by Eimeria parasites, is responsible for major losses in the poultry industry by impacting chicken health. The gut microbiota is associated with health factors, such as nutrient exchange and immune system modulation, requiring understanding on the effects of Eimeria infection on the gut microbiota. This study aimed to determine the effects of Eimeria acervulina infection on the luminal and mucosal microbiota of the cecum (CeL and CeM) and ileum (IlL and IlM) at multiple time points (days 3, 5, 7, 10, and 14) post-infection. E. acervulina infection decreased evenness in CeL microbiota at day 10, increased richness in CeM microbiota at day 3 before decreasing richness at day 14, and decreased richness in IlL microbiota from day 3 to 10. CeL, CeM, and IlL microbiota differed between infected and control birds based on beta diversity at varying time points. Infection reduced relative abundance of bacterial taxa and some predicted metabolic pathways known for short-chain fatty acid production in CeL, CeM, and IlL microbiota, but further understanding of metabolic function is required. Despite E. acervulina primarily targeting the duodenum, our findings demonstrate the infection can impact bacterial diversity and abundance in the cecal and ileal microbiota.
Assuntos
Ceco , Galinhas , Coccidiose , Eimeria , Microbioma Gastrointestinal , Íleo , Doenças das Aves Domésticas , Animais , Galinhas/microbiologia , Galinhas/parasitologia , Ceco/microbiologia , Ceco/parasitologia , Eimeria/fisiologia , Íleo/microbiologia , Íleo/parasitologia , Coccidiose/veterinária , Coccidiose/parasitologia , Doenças das Aves Domésticas/microbiologia , Doenças das Aves Domésticas/parasitologia , Mucosa Intestinal/microbiologia , Mucosa Intestinal/parasitologiaRESUMO
BACKGROUND: Cryptosporidium spp. cause watery diarrhea in humans and animals, especially in infants and neonates. They parasitize the apical surface of the epithelial cells in the intestinal lumen. However, the pathogenesis of Cryptosporidium-induced diarrhea is not fully understood yet. METHODOLOGY/PRINCIPAL FINDINGS: In this study, we infected C57BL/6j neonatal mice with C. parvum IIa and IId subtypes, and examined oocyst burden, pathological changes, and intestinal epithelial permeability during the infection. In addition, transcriptomic analyses were used to study the mechanism of diarrhea induced by the C. parvum IId subtype. The neonatal mice were sensitive to both C. parvum IIa and IId infection, but the IId subtype caused a wide oocyst shedding window and maintained the high oocyst burden in the mice compared with the IIa subtype. In addition, the mice infected with C. parvum IId resulted in severe intestinal damage at the peak of infection, leading to increased permeability of the epithelial barrier. The KEGG, GO and GSEA analyses revealed that the downregulation of adherens junction and cell junction molecules at 11 dpi. Meanwhile, E-cadherin, which is associated with adherens junction, was reduced at the protein level in mouse ileum at peak and late infection. CONCLUSIONS/SIGNIFICANCE: C. parvum IId infection causes more severe pathological damage than C. parvum IIa infection in neonatal mice. Furthermore, the impairment of the epithelial barrier during C. parvum IId infection results from the downregulation of intestinal junction proteins.
Assuntos
Animais Recém-Nascidos , Criptosporidiose , Cryptosporidium parvum , Regulação para Baixo , Mucosa Intestinal , Camundongos Endogâmicos C57BL , Animais , Cryptosporidium parvum/genética , Criptosporidiose/parasitologia , Criptosporidiose/patologia , Camundongos , Mucosa Intestinal/parasitologia , Mucosa Intestinal/patologia , Caderinas/metabolismo , Caderinas/genética , Diarreia/parasitologia , Células Epiteliais/parasitologia , Feminino , Oocistos , Íleo/parasitologia , Íleo/patologia , Modelos Animais de DoençasRESUMO
Leishmania (Viannia) braziliensis is one of the main causes of cutaneous leishmaniasis in the Americas. This species presents genetic polymorphism that can cause destructive lesions in oral, nasal, and oropharyngeal tracts. In a previous study, the parasite caused several histopathological changes to hamster ileums. Our study evaluates immune response components, morphological changes, and effects on neurons in the ileums of hamsters infected by three different strains of L. (V.) braziliensis in two infection periods. For the experiment, we separated hamsters into four groups: a control group and three infected groups. Infected hamsters were euthanized 90- or 120-days post infection. We used three strains of L. (V.) braziliensis: the reference MHOM/BR/1975/M2903 and two strains isolated from patients who had different responses to Glucantime® treatment (MHOM/BR/2003/2314 and MHOM/BR/2000/1655). After laparotomy, ileums were collected for histological processing, biochemical analysis, and evaluation of neurons in the myenteric and submucosal plexuses of the enteric nervous system (ENS). The results demonstrated the increase of blood leukocytes after the infection. Optical microscopy analysis showed histopathological changes with inflammatory infiltrates, edemas, ganglionitis, and Leishmania amastigotes in the ileums of infected hamsters. We observed changes in the organ histoarchitecture of infected hamsters when compared to control groups, such as thicker muscular and submucosa layers, deeper and wider crypts, and taller and broader villi. The number of intraepithelial lymphocytes and TGF-ß-immunoreactive cells increased in all infected groups when compared to the control groups. Mast cells increased with longer infection periods. The infection also caused remodeling of intestinal collagen and morphometry of myenteric and submucosal plexus neurons; but this effect was dependent on infection duration. Our results show that L. (V.) braziliensis infection caused time-dependent alterations in hamster ileums. This was demonstrated by the reduction of inflammatory cells and the increase of tissue regeneration factors at 120 days of infection. The infected groups demonstrated different profiles in organ histoarchitecture, migration of immune cells, and morphometry of ENS neurons. These findings suggest that the small intestine (or at least the ileum) is a target organ for L. (V.) braziliensis infection, as the infection caused changes that were dependent on duration and strain.
Assuntos
Íleo/parasitologia , Leishmania braziliensis , Leishmaniose/patologia , Animais , Cricetinae , HumanosRESUMO
Cryptosporidiosis is a gastrointestinal illness with profuse diarrhoea. Although there are no other Food and Drug Administration (FDA)-approved alternatives for the treatment of cryptosporidiosis, nitazoxanide (NTZ) can be qualified as partially effective. In immunosuppressed conditions, severe and/or disseminated cryptosporidiosis may occur and patients should be treated parenterally. To achieve the goal of developing parenteral treatment for cryptosporidiosis, the current study was undertaken to investigate the in vitro and in vivo anticryptosporidial activity of aminoxanide. This new l-tert-leucyl thiazolide is a soluble prodrug of tizoxanide (TIZ), the main metabolite of NTZ. Confirming the good efficacy of aminoxanide in Cryptosporidium parvum-infected HCT-8 cells with a 50% inhibitory concentration of 1.55 µm (±0.21), in immunosuppressed C. parvum-infected Mongolian gerbils (Meriones unguiculatus), a 5-day treatment with a daily intramuscular dose of 100 mg kg−1 aminoxanide resulted in a 72.5% oocyst excretion inhibition, statistically equivalent to 75.5% in gerbils treated with a 4-fold lower oral dose of NTZ. Cryptosporidium parvum-induced intestinal pathology and inflammation were improved. Aminoxanide provides an injectable form of TIZ that NTZ was unable to do and is a promising drug for which optimization of the formulation should be further explored. These results represent a first promising step towards the goal of developing a parenteral treatment for cryptosporidiosis.
Assuntos
Criptosporidiose/tratamento farmacológico , Cryptosporidium parvum/efeitos dos fármacos , Ésteres/uso terapêutico , Pró-Fármacos/uso terapêutico , Tiazóis/uso terapêutico , Animais , Bovinos , Linhagem Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Ésteres/química , Ésteres/farmacologia , Fezes/parasitologia , Feminino , Gerbillinae , Íleo/parasitologia , Íleo/patologia , Concentração Inibidora 50 , Masculino , Estrutura Molecular , Pró-Fármacos/química , Pró-Fármacos/farmacologia , Tiazóis/química , Tiazóis/farmacologiaRESUMO
Introduction. Cryptosporidium parvum causes intestinal parasitic infections affecting both immunosuppressed and immunocompetent individuals.Gap statement. Given the absence of effective treatments for cryptosporidiosis, especially in immunodeficient patients, the present study was designed to assess the therapeutic efficacy of secnidazole (SEC) and its combination with nitazoxanide (NTZ) in comparison to single NTZ treatment in relation to the immune status of a murine model of C. parvum infection.Methodology. The infected groups were administered NTZ, SEC or NTZ-SEC for three or five successive doses. At days 10 and 12 post-infection (p.i.), the mice were sacrificed, and the efficacy of the applied drugs was evaluated by comparing the histopathological alterations in ileum and measuring the T helper Th1 (interferon gamma; IFN-γ), Th2 [interleukin (IL)-4 and IL-10] and Th17 (IL-17) cytokine profiles in serum.Results. The NTZ-SEC combination recorded the maximal reduction of C. parvum oocyst shedding, endogenous stages count and intestinal histopathology, regardless of the immune status of the infected mice. The efficacy of NTZ-SEC was dependent on the period of administration, as the 5 day-based treatment protocol was also more effective than the 3 day-based one in terms of immunocompetence and immunosuppression. The present treatment schedule induced an immunomodulatory effect from SEC that developed a protective immune response against C. parvum infection with reduced production of serum IL-17, IFN-γ, IL-4 and IL-10.Conclusions. Application of NTZ-SEC combined therapy may be useful in treatment of C. parvum, especially in cases involving immunosuppression.
Assuntos
Antiprotozoários/uso terapêutico , Criptosporidiose/tratamento farmacológico , Imunomodulação/efeitos dos fármacos , Metronidazol/análogos & derivados , Nitrocompostos/uso terapêutico , Tiazóis/uso terapêutico , Animais , Criptosporidiose/imunologia , Criptosporidiose/parasitologia , Criptosporidiose/patologia , Cryptosporidium parvum/efeitos dos fármacos , Citocinas/sangue , Modelos Animais de Doenças , Esquema de Medicação , Quimioterapia Combinada , Íleo/efeitos dos fármacos , Íleo/parasitologia , Íleo/patologia , Hospedeiro Imunocomprometido , Masculino , Metronidazol/uso terapêutico , Camundongos , Carga ParasitáriaRESUMO
AIMS. GIARDIA: is sometimes missed by the pathologist, and we sought to determine how often this occurs at our institution-a large tertiary care center with a subspecialty gastrointestinal pathology service and what certain clinical and histologic clues can be used to flag cases with a higher likelihood of infection, targeting them for greater scrutiny. METHODS AND RESULTS: We identified a set of patients who tested positive for Giardia with a stool-based test, and who also received a small bowel biopsy at a similar time-point. These biopsies were retrospectively reviewed for Giardia, finding 8 positive cases. The organism was prospectively detected in 4 cases (50%) but overlooked in the remaining 4 cases (50%). Three of the 4 cases missed cases showed only rare organisms. The detected cases tended to more frequently have prominent lymphoid aggregates (3 detected cases, 0 overlooked cases) and intraepithelial lymphocytosis (3 detected cases, 0 overlooked cases). Certain clinical and histologic clues can be used to flag cases with a higher likelihood of infection. Specifically, we found abnormalities of the mucosa (active inflammation, intraepithelial lymphocytosis, villous expansion, prominent lymphoid aggregates) in each case, and 4 of 8 cases were from immunocompromised patients. Finally, 2 of 8 cases were terminal ileum biopsies. CONCLUSIONS: Biopsies with a histologic abnormality or those from immunocompromised patients should receive greater attention. Routinely looking for Giardia at that terminal ileum is necessary.
Assuntos
Duodeno/parasitologia , Giardia/isolamento & purificação , Giardíase/diagnóstico , Íleo/parasitologia , Mucosa Intestinal/parasitologia , Adulto , Idoso , Biópsia , Pré-Escolar , Duodeno/imunologia , Duodeno/patologia , Fezes/parasitologia , Feminino , Giardia/imunologia , Giardíase/imunologia , Giardíase/parasitologia , Giardíase/patologia , Hospitais com Alto Volume de Atendimentos , Humanos , Íleo/imunologia , Íleo/patologia , Hospedeiro Imunocomprometido , Mucosa Intestinal/patologia , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Centros de Atenção Terciária , Adulto JovemRESUMO
Malaria strongly predisposes to bacteremia, which is associated with sequestration of parasitized red blood cells and increased gastrointestinal permeability. The mechanisms underlying this disruption are poorly understood. Here, we evaluated the expression of factors associated with mast cell activation and malaria-associated bacteremia in a rodent model. C57BL/6J mice were infected with Plasmodium yoeliiyoelli 17XNL, and blood and tissues were collected over time to assay for circulating levels of bacterial 16S DNA, IgE, mast cell protease 1 (Mcpt-1) and Mcpt-4, Th1 and Th2 cytokines, and patterns of ileal mastocytosis and intestinal permeability. The anti-inflammatory cytokines (interleukin-4 [IL-4], IL-6, and IL-10) and MCP-1/CCL2 were detected early after P. yoeliiyoelii 17XNL infection. This was followed by the appearance of IL-9 and IL-13, cytokines known for their roles in mast cell activation and growth-enhancing activity as well as IgE production. Later increases in circulating IgE, which can induce mast cell degranulation, as well as Mcpt-1 and Mcpt-4, were observed concurrently with bacteremia and increased intestinal permeability. These results suggest that P. yoeliiyoelii 17XNL infection induces the production of early cytokines that activate mast cells and drive IgE production, followed by elevated IgE, IL-9, and IL-13 that maintain and enhance mast cell activation while disrupting the protease/antiprotease balance in the intestine, contributing to epithelial damage and increased permeability.
Assuntos
Bacteriemia/imunologia , Citocinas/sangue , Malária/imunologia , Mastócitos/metabolismo , Plasmodium yoelii/imunologia , Animais , Bacteriemia/parasitologia , Quimiocina CCL2/sangue , Quimases/sangue , Feminino , Íleo/citologia , Íleo/metabolismo , Íleo/parasitologia , Imunoglobulina E/sangue , Inflamação/sangue , Interleucina-10/sangue , Interleucina-13/metabolismo , Interleucina-4/sangue , Interleucina-6/sangue , Interleucina-9/sangue , Leucócitos/citologia , Malária/sangue , Malária/parasitologia , Camundongos , Camundongos Endogâmicos C57BL , Permeabilidade , RNA Ribossômico 16S/sangue , RNA Ribossômico 16S/genéticaRESUMO
BACKGROUND: Echinostoma caproni (Trematoda: Echinostomatidae) is an intestinal trematode, which has been extensively used to investigate the factors that determine the rejection of intestinal helminths. In this sense, several studies have shown that IL-25 is critical for the development of resistance against E. caproni in mice. In fact, treatment of mice with recombinant IL-25 generates resistance against primary E. caproni infection. However, the mechanisms by which IL-25 induces resistance remain unknown. METHODS: To study the mechanisms responsible for resistance elicited by IL-25, we analyzed the ileal proteomic changes induced by IL-25 in mice and their potential role in resistance. To this purpose, we compared the protein expression profiles in the ileum of four experimental groups of mice: naïve controls; E. caproni-infected mice; rIL-25-treated mice; and rIL-25-treated mice exposed to E. caproni metacercariae. RESULTS: Quantitative comparison by 2D-DIGE showed significant changes in a total of 41 spots. Of these, 40 validated protein spots were identified by mass spectrometry corresponding to 24 proteins. CONCLUSIONS: Our results indicate that resistance to infection is associated with the maintenance of the intestinal epithelial homeostasis and the regulation of proliferation and cell death. These results provide new insights into the proteins involved in the regulation of tissue homeostasis after intestinal infection and its transcendence in resistance.
Assuntos
Echinostoma/patogenicidade , Equinostomíase , Íleo/metabolismo , Interleucinas/metabolismo , Proteínas/metabolismo , Animais , Anti-Helmínticos/farmacologia , Morte Celular , Proliferação de Células , Equinostomíase/parasitologia , Equinostomíase/prevenção & controle , Íleo/parasitologia , Mucosa Intestinal/metabolismo , Mucosa Intestinal/parasitologia , Espectrometria de Massas , Camundongos , Praziquantel/farmacologia , ProteômicaRESUMO
Pseudocorynosoma constrictum (Van Cleave, 1918) is a polymorphid acanthocephalan that attaches to the digestive tract of waterfowl to complete its life cycle, causing severe histological damage to its definitive avian hosts. In the present study, we present a histopathological analysis of the lesions that P. constrictum induced in the layers of the ileum of the blue-winged teal Anas discors. The results revealed that worms insert the attachment structures into the inner gut muscular layer, which causes substantial swelling, haemorrhaging and necrosis in the tissue near the parasite's proboscis. We also observed that the number of parasites attached to the tissue can obstruct the intestinal lumen; in the most serious case, we observed more than 30 parasites penetrating completely the walls of the bird intestine.
Assuntos
Acantocéfalos/patogenicidade , Aves/parasitologia , Íleo/patologia , Íleo/parasitologia , Acantocéfalos/anatomia & histologia , Animais , México , Mucosa/parasitologia , NecroseRESUMO
The gastrointestinal helminth parasites of 170 common wallaroos or euros, Osphranter robustus (Gould), collected from all mainland states in which the species occurs as well as the Northern Territory, are presented, including previously published data. A total of 65 species of helminths were encountered, including four species of anoplocephalid cestodes found in the bile ducts and small intestine, and 61 species of strongylid nematodes, all but two of which occurring in the stomach, and with the remainder occurring in the terminal ileum, caecum and colon. Among the mainland subspecies of O. robustus, 52 species of helminths were encountered in O. r. robustus, compared with 30 species in O. r. woodwardi and 35 species in O. r. erubescens. Of the parasite species encountered, only 17 were specific to O. robustus, the remaining being shared with sympatric host species. Host-specific species or species occurring in O. robustus at a high prevalence can be classified as follows: widely distributed; restricted to northern Australia; restricted to the northern wallaroo, O. r. woodwardi; found only in the euro, O. r. erubescens; found essentially along the eastern coast of Australia, primarily in O. r. robustus; and species with highly limited regional distributions. The data currently available suggest that the acquisition of a significant number of parasites is due to co-grazing with other macropodids, while subspeciation in wallaroos as well as climatic variables may have influenced the diversification of the parasite fauna.
Assuntos
Helmintíase , Helmintos/isolamento & purificação , Intestinos/parasitologia , Macropodidae/parasitologia , Infecções por Strongylida/veterinária , Distribuição Animal , Animais , Austrália/epidemiologia , Ductos Biliares/parasitologia , Biodiversidade , Cestoides/isolamento & purificação , Cestoides/parasitologia , Colo/parasitologia , Helmintíase/parasitologia , Helmintíase/transmissão , Helmintos/parasitologia , Especificidade de Hospedeiro , Íleo/parasitologia , Enteropatias Parasitárias/veterinária , Nematoides/isolamento & purificação , Nematoides/parasitologia , Estômago/parasitologia , Estrongilídios/isolamento & purificação , Estrongilídios/parasitologia , Infecções por Strongylida/parasitologia , Infecções por Strongylida/transmissãoRESUMO
Tuft cells are an epithelial cell type critical for initiating type 2 immune responses to parasites and protozoa in the small intestine. To respond to these stimuli, intestinal tuft cells use taste chemosensory signaling pathways, but the role of taste receptors in type 2 immunity is poorly understood. In this study, we show that the taste receptor TAS1R3, which detects sweet and umami in the tongue, also regulates tuft cell responses in the distal small intestine. BALB/c mice, which have an inactive form of TAS1R3, as well as Tas1r3-deficient C57BL6/J mice both have severely impaired responses to tuft cell-inducing signals in the ileum, including the protozoa Tritrichomonas muris and succinate. In contrast, TAS1R3 is not required to mount an immune response to the helminth Heligmosomoides polygyrus, which infects the proximal small intestine. Examination of uninfected Tas1r3-/- mice revealed a modest reduction in the number of tuft cells in the proximal small intestine but a severe decrease in the distal small intestine at homeostasis. Together, these results suggest that TAS1R3 influences intestinal immunity by shaping the epithelial cell landscape at steady-state.
Assuntos
Células Epiteliais/imunologia , Mucosa Intestinal/imunologia , Intestino Delgado/imunologia , Receptores Acoplados a Proteínas G/imunologia , Receptores Acoplados a Proteínas G/metabolismo , Animais , Células Epiteliais/metabolismo , Microbioma Gastrointestinal , Homeostase , Íleo/imunologia , Íleo/parasitologia , Mucosa Intestinal/metabolismo , Mucosa Intestinal/parasitologia , Intestino Delgado/parasitologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Nematospiroides dubius/imunologia , Receptores Acoplados a Proteínas G/deficiência , Infecções por Strongylida/imunologia , Infecções por Strongylida/parasitologia , Paladar/fisiologia , Tritrichomonas/imunologiaAssuntos
Anisaquíase , Anisakis/isolamento & purificação , Colonoscopia/métodos , Produtos Pesqueiros/efeitos adversos , Íleo , Alimentos Crus/efeitos adversos , Idoso , Animais , Anisaquíase/diagnóstico , Anisaquíase/etiologia , Anisaquíase/fisiopatologia , Anisaquíase/terapia , Humanos , Íleo/diagnóstico por imagem , Íleo/parasitologia , Achados Incidentais , Masculino , Resultado do Tratamento , Zoonoses/diagnóstico , Zoonoses/etiologia , Zoonoses/fisiopatologia , Zoonoses/prevenção & controleRESUMO
Cryptosporidiosis threatens life of young children in developing countries and newborn calves around the world. No vaccine or therapy can prevent or cure this diarrhea-inducing enteric disease caused by Cryptosporidium spp. protozoan parasites. There is an essential need to discover new therapeutic drugs efficient in reducing parasite burden in infected individuals. Research therefore relies on reliable small animal models of cryptosporidiosis. Here, we present excellent mouse models which can efficiently mimic pathogenesis of human and bovine cryptosporidiosis. We also describe methods to purify C. parvum oocysts from stool and intestine of infected mice to facilitate oocyst quantification. Moreover, we present protocols using flow cytometry, quantitative polymerase chain reaction, and histopathology to accurately quantify parasite burden in stool or intestine samples.
Assuntos
Criptosporidiose/parasitologia , Cryptosporidium parvum/isolamento & purificação , Oocistos/isolamento & purificação , Animais , Criptosporidiose/patologia , Cryptosporidium , Cryptosporidium parvum/crescimento & desenvolvimento , Modelos Animais de Doenças , Fezes/parasitologia , Citometria de Fluxo/métodos , Íleo/citologia , Íleo/parasitologia , Íleo/patologia , Interleucina-12/genética , Camundongos , Camundongos Knockout , Camundongos SCID , Oocistos/crescimento & desenvolvimento , Reação em Cadeia da Polimerase em Tempo Real/métodos , Receptores de Interferon/genética , Fluxo de Trabalho , Receptor de Interferon gamaRESUMO
Cryptosporidium is an intestinal pathogen that causes severe but self-limiting diarrhea in healthy humans, yet it can turn into a life-threatening, unrelenting infection in immunocompromised patients and young children. Severe diarrhea is recognized as the leading cause of mortality for children below 5 years of age in developing countries. The only approved treatment against cryptosporidiosis, nitazoxanide, has limited efficacy in the most vulnerable patient populations, including malnourished children, and is ineffective in immunocompromised individuals. Here, we investigate inhibition of the parasitic cleavage and polyadenylation specificity factor 3 (CPSF3) as a strategy to control Cryptosporidium infection. We show that the oxaborole AN3661 selectively blocked Cryptosporidium growth in human HCT-8 cells, and oral treatment with AN3661 reduced intestinal parasite burden in both immunocompromised and neonatal mouse models of infection with greater efficacy than nitazoxanide. Furthermore, we present crystal structures of recombinantly produced Cryptosporidium CPSF3, revealing a mechanism of action whereby the mRNA processing activity of this enzyme is efficiently blocked by the binding of the oxaborole group at the metal-dependent catalytic center. Our data provide insights that may help accelerate the development of next-generation anti-Cryptosporidium therapeutics.
Assuntos
Fator de Especificidade de Clivagem e Poliadenilação/metabolismo , Criptosporidiose/genética , Criptosporidiose/parasitologia , Cryptosporidium/genética , Metais/química , Precursores de RNA/genética , Processamento Pós-Transcricional do RNA , Animais , Antiparasitários/química , Antiparasitários/farmacologia , Linhagem Celular Tumoral , Fator de Especificidade de Clivagem e Poliadenilação/química , Cristalização , Humanos , Íleo/parasitologia , Íleo/ultraestrutura , Camundongos Endogâmicos C57BL , Modelos Moleculares , Proteínas Recombinantes/metabolismoRESUMO
The protozoan parasite Cryptosporidium parvum (CP) causes cryptosporidiosis, a diarrheal disease worldwide. Infection in immunocompetent hosts typically results in acute, self-limiting, or recurrent diarrhea. However, in immunocompromised individuals infection can cause fulminant diarrhea, extraintestinal manifestations, and death. To date, the mechanisms underlying CP-induced diarrheal pathogenesis are poorly understood. Diarrheal diseases most commonly involve increased secretion and/or decreased absorption of fluid and electrolytes. We and others have previously shown impaired chloride absorption in infectious diarrhea due to dysregulation of SLC26A3 [downregulated in adenoma (DRA)], the human intestinal apical membrane Cl-/HCO3- exchanger protein. However, there are no studies on the effects of CP infection on DRA activity. Therefore, we examined the expression and function of DRA in intestinal epithelial cells in response to CP infection in vitro and in vivo. CP infection (0.5 × 106 oocysts/well in 24-well plates, 24 h) of Caco-2 cell monolayers significantly decreased Cl-/HCO3- exchange activity (measured as DIDS-sensitive 125I uptake) as well as DRA mRNA and protein levels. Substantial downregulation of DRA mRNA and protein was also observed following CP infection ex vivo in mouse enteroid-derived monolayers and in vivo in the ileal and jejunal mucosa of C57BL/6 mice for 24 h. However, at 48 h after infection in vivo, the effects on DRA mRNA and protein were attenuated and at 5 days after infection DRA returned to normal levels. Our results suggest that impaired chloride absorption due to downregulation of DRA could be one of the contributing factors to CP-induced acute, self-limiting diarrhea in immunocompetent hosts.
Assuntos
Antiporters/genética , Antiportadores de Cloreto-Bicarbonato/genética , Criptosporidiose/genética , Cryptosporidium parvum/patogenicidade , Regulação da Expressão Gênica/genética , Mucosa Intestinal/metabolismo , Transportadores de Sulfato/genética , Animais , Anticorpos Neutralizantes/farmacologia , Antiporters/antagonistas & inibidores , Antiporters/metabolismo , Células CACO-2 , Antiportadores de Cloreto-Bicarbonato/antagonistas & inibidores , Antiportadores de Cloreto-Bicarbonato/metabolismo , Cloretos/metabolismo , Criptosporidiose/metabolismo , Criptosporidiose/parasitologia , Cryptosporidium parvum/fisiologia , Interações Hospedeiro-Parasita/genética , Humanos , Íleo/metabolismo , Íleo/parasitologia , Mucosa Intestinal/parasitologia , Transporte de Íons , Janus Quinases/genética , Janus Quinases/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Organoides/metabolismo , Organoides/parasitologia , Fatores de Transcrição STAT/genética , Fatores de Transcrição STAT/metabolismo , Transportadores de Sulfato/antagonistas & inibidores , Transportadores de Sulfato/metabolismoRESUMO
Feeder organelles of Cryptosporidium are the convoluted structures located at the host-parasite interface that uptake of nutrients from host cells. Although the ultrastructure of feeder organelles has been summarized as being highly invaginated structure, the three-dimensional form remains uncertain. Osmium-maceration scanning electron microscopy (OS-SEM) allows visualization of the three-dimensional ultrastructure after removing soluble proteins. Here, we assessed C. parvum attached to mouse ileal epithelial cells using transmission electron microscopy (TEM) and OS-SEM. Feeder organelles visualized by TEM as aggregated structures of concentrically-, vertically- and randomly-lined bars comprised a complex reticulated network of stacked flat bursiform, ring-shaped bursiform and reticulated tubular membranes on OS-SEM. These findings suggested that the feeder organelles are more complex than was previously thought.
Assuntos
Cryptosporidium parvum/ultraestrutura , Células Epiteliais/parasitologia , Íleo/parasitologia , Animais , Camundongos , Camundongos SCID , Microscopia Eletrônica de Varredura , Microscopia Eletrônica de Transmissão , Organelas/ultraestrutura , OsmioRESUMO
Oral T. gondii infection (30 cysts of 76K strain) induces acute lethal ileitis in sensitive C57BL/6 (B6) mice with increased expression of IL-33 and its receptor ST2 in the ileum. Here we show that IL-33 is involved in ileitis, since absence of IL-33R/ST2 attenuated neutrophilic inflammation and Th1 cytokines upon T. gondii infection with enhanced survival. Blockade of ST2 by neutralizing ST2 antibody in B6 mice conferred partial protection, while rmIL-33 aggravated ileitis. Since IL-22 expression further increased in absence of ST2, we blocked IL-22 by neutralizing antibody, which abrogated protection from acute ileitis in ST2 deficient mice. In conclusion, severe lethal ileitis induced by oral T. gondii infection is attenuated by blockade of ST2 signaling and may be mediated in part by endogenous IL-22.
Assuntos
Proteína 1 Semelhante a Receptor de Interleucina-1/metabolismo , Interleucinas/metabolismo , Toxoplasma/metabolismo , Toxoplasmose Animal/metabolismo , Animais , Citocinas/metabolismo , Microbioma Gastrointestinal/fisiologia , Ileíte/metabolismo , Ileíte/parasitologia , Íleo/metabolismo , Íleo/parasitologia , Inflamação/metabolismo , Inflamação/parasitologia , Interferon gama/metabolismo , Mucosa Intestinal/metabolismo , Mucosa Intestinal/parasitologia , Camundongos , Camundongos Endogâmicos C57BL , Transdução de Sinais/fisiologia , Interleucina 22RESUMO
We have recently shown that following peroral low-dose Toxoplasma gondii infection susceptible mice develop subacute ileitis within 10 days. Data regarding long-term intestinal and extra-intestinal sequelae of infection are scarce, however. We therefore challenged conventional C57BL/6 mice with one cyst of T. gondii ME49 strain by gavage and performed a comprehensive immunopathological survey 10, 36, and 57 days later. As early as 10 days post-infection, mice were suffering from subacute ileitis as indicated by mild-to-moderate histopathological changes of the ileal mucosa. Furthermore, numbers of apoptotic and proliferating/regenerating epithelial cells as well as of T and B lymphocytes in the mucosa and lamina propria of the ileum were highest at day 10 post-infection, but declined thereafter, and were accompanied by enhanced pro-inflammatory mediator secretion in ileum, colon and mesenteric lymph nodes that was most pronounced during the early phase of infection. In addition, subacute ileitis was accompanied by distinct shifts in the commensal gut microbiota composition in the small intestines. Remarkably, immunopathological sequelae of T. gondii infection were not restricted to the intestines, but could also be observed in extra-intestinal tissues including the liver, kidneys, lungs, heart and strikingly, in systemic compartments that were most prominent at day 10 post-infection. We conclude that the here provided long-term kinetic survey of immunopathological sequalae following peroral low-dose T. gondii infection provides valuable corner stones for a better understanding of the complex interactions within the triangle relationship of (parasitic) pathogens, the host immunity and the commensal gut microbiota during intestinal inflammation. The low-dose T. gondii infection model may be applied as valuable gut inflammation model in future pre-clinical studies in order to test potential treatment options for intestinal inflammatory conditions in humans.
Assuntos
Ileíte/patologia , Toxoplasma/crescimento & desenvolvimento , Toxoplasmose Animal/patologia , Estruturas Animais/parasitologia , Estruturas Animais/patologia , Animais , Modelos Animais de Doenças , Disbiose , Microbioma Gastrointestinal , Histocitoquímica , Ileíte/complicações , Ileíte/parasitologia , Íleo/parasitologia , Íleo/patologia , Mucosa Intestinal/parasitologia , Mucosa Intestinal/patologia , Estudos Longitudinais , Camundongos Endogâmicos C57BL , Parasitemia/parasitologia , Parasitemia/patologia , Toxoplasmose Animal/parasitologiaRESUMO
The parasitic helminth Trichinella spiralis, which poses a serious health risk to animals and humans, can be found worldwide. Recent findings indicate that a rare type of gut epithelial cell, tuft cells, can detect the helminth, triggering type 2 immune responses. However, the underlying molecular mechanisms remain to be fully understood. Here we show that both excretory-secretory products (E-S) and extract of T. spiralis can stimulate the release of the cytokine interleukin 25 (IL-25) from the mouse small intestinal villi and evoke calcium responses from tuft cells in the intestinal organoids, which can be blocked by a bitter-taste receptor inhibitor, allyl isothiocyanate. Heterologously expressed mouse Tas2r bitter-taste receptors, the expression of which is augmented during tuft-cell hyperplasia, can respond to the E-S and extract as well as to the bitter compound salicin whereas salicin in turn can induce IL-25 release from tuft cells. Furthermore, abolishment of the G-protein γ13 subunit, application of the inhibitors for G-protein αo/i, Gßγ subunits, and phospholipase Cß2 dramatically reduces the IL-25 release. Finally, tuft cells are found to utilize the inositol triphosphate receptor type 2 (Ip3r2) to regulate cytosolic calcium and thus Trpm5 activity, while potentiation of Trpm5 by a sweet-tasting compound, stevioside, enhances tuft cell IL-25 release and hyperplasia in vivo. Taken together, T. spiralis infection activates a signaling pathway in intestinal tuft cells similar to that of taste-bud cells, but with some key differences, to initiate type 2 immunity.
Assuntos
Intestino Delgado/parasitologia , Transdução de Sinais , Trichinella spiralis , Triquinelose/metabolismo , Animais , Duodeno/citologia , Duodeno/metabolismo , Duodeno/parasitologia , Antígenos de Histocompatibilidade Classe II , Íleo/citologia , Íleo/metabolismo , Íleo/parasitologia , Interleucina-17/metabolismo , Intestino Delgado/citologia , Intestino Delgado/metabolismo , Jejuno/citologia , Jejuno/metabolismo , Jejuno/parasitologia , Camundongos , Triquinelose/parasitologiaRESUMO
Toxoplasma gondii is a common perorally transmitted parasite; however, its immunopathogenesis in gut-associated tissues remains unclear. Here, we compared disease manifestation in C57BL/6 immunocompetent wild type (WT) mice and immunocompromised interferon (IFN)-γ-deficient (GKO) mice after peroral infection (PI) with T. gondii cysts (Fukaya strain). Strong PI-induced Th1 cytokine expression was detected in WT mice. Moreover, bradyzoite-specific T.g.HSP30/bag1 mRNA was detected in the ileum parenchyma and Peyer's patches (PP), but not in the mesenteric lymph nodes, at 7â¯days post-infection in WT mice, and was significantly higher than that in GKO mice. Nested PCR showed that parasites existed in ileum parenchyma at days 1 and 1.5 post-PI in GKO and WT mice, respectively. In addition, quantitative competitive-PCR indicated that T. gondii first colonized the PP (day 3 post-PI), followed by the ileum parenchyma and mesenteric lymph nodes, spleen, and portal and aortic blood (day 7 post-PI). Although parasites were consistently more abundant in GKO mice, similar invasion and dissemination patterns were observed in the two hosts. Collectively, these data suggest that some zoites differentiate from tachyzoites to bradyzoites in the ileum and that T. gondii initially invades the ileum parenchyma, and then accumulates and proliferates in the PP before disseminating through the lymphatic systems of both GKO and WT hosts.