RESUMEN
This paper centers on some whole-istic organizational and functional aspects of hepatic Schistosoma mansoni granuloma, which is an extremely complex system. First, it structurally develops a collagenic topology, originated bidirectionally from an inward and outward assembly of growth units. Inward growth appears to be originated from myofibroblasts derived from small portal vessel around intravascular entrapped eggs, while outward growth arises from hepatic stellate cells. The auto-assembly of the growth units defines the three-dimensional scaffold of the schistosome granulomas. The granuloma surface irregularity and its border presented fractal dimension equal to 1.58. Second, it is internally regulated by intricate networks of immuneneuroendocrine stimuli orchestrated by leptin and leptin receptors, substance P and Vasoactive intestinal peptide. Third, it can reach the population of ± 40,000 cells and presents an autopoietic component evidenced by internal proliferation (Ki-67+ Cells), and by expression of c-Kit+ Cells, leptin and leptin receptor (Ob-R), granulocyte-colony stimulating factor (G-CSF-R), and erythropoietin (Epo-R) receptors. Fourth, the granulomas cells are intimately connected by pan-cadherins, occludin and connexin-43, building a state of closing (granuloma closure). In conclusion, the granuloma is characterized by transitory stages in such a way that its organized structure emerges as a global property which is greater than the sum of actions of its individual cells and extracellular matrix components.
Asunto(s)
Animales , Ratones , Granuloma/patología , Parasitosis Hepáticas/patología , Schistosoma mansoni/fisiología , Esquistosomiasis mansoni/patología , Modelos Animales de Enfermedad , Técnica del Anticuerpo Fluorescente Indirecta , Fractales , Granuloma/metabolismo , Granuloma/parasitología , Parasitosis Hepáticas/metabolismo , Parasitosis Hepáticas/parasitología , Coloración y Etiquetado , Esquistosomiasis mansoni/metabolismo , Esquistosomiasis mansoni/parasitología , Factores de TiempoRESUMEN
We have previously showed that Schistosoma mansoni ATP-diphosphohydrolase and Solanum tuberosum potato apyrase share epitopes and the vegetable protein has immunostimulatory properties. Here, it was verified the in situ cross-immunoreactivity between mice NTPDases and anti-potato apyrase antibodies produced in rabbits, using confocal microscopy. Liver samples were taken from Swiss Webster mouse 8 weeks after infection with S. mansoni cercariae, and anti-potato apyrase and TRITC-conjugated anti-rabbit IgG antibody were tested on cryostat sections. The results showed that S. mansoni egg ATP diphosphohydrolase isoforms, developed by anti-potato apyrase, are expressed in miracidial and egg structures, and not in granulomatous cells and hepatic structures (hepatocytes, bile ducts, and blood vessels). Therefore, purified potato apyrase when inoculated in rabbit generates polyclonal sera containing anti-apyrase antibodies that are capable of recognizing specifically S. mansoni ATP diphosphohydrolase epitopes, but not proteins from mammalian tissues, suggesting that autoantibodies are not induced during potato apyrase immunization. A phylogenetic tree obtained for the NTPDase family showed that potato apyrase had lower homology with mammalian NTPDases 1-4, 7, and 8. Further analysis of potato apyrase epitopes could implement their potential use in schistosomiasis experimental models.
Asunto(s)
Animales , Masculino , Ratones , Conejos , Adenosina Trifosfatasas/inmunología , Apirasa/inmunología , Schistosoma mansoni/enzimología , Esquistosomiasis mansoni/inmunología , Solanum tuberosum/enzimología , Secuencia de Aminoácidos , Adenosina Trifosfatasas/metabolismo , Anticuerpos Antihelmínticos/inmunología , Apirasa/metabolismo , Reacciones Cruzadas , Modelos Animales de Enfermedad , Microscopía Confocal , Datos de Secuencia Molecular , Schistosoma mansoni/inmunología , Schistosoma mansoni/metabolismoRESUMEN
This paper centers on some whole-istic organizational and functional aspects of hepatic Schistosoma mansoni granuloma, which is an extremely complex system. First, it structurally develops a collagenic topology, originated bidirectionally from an inward and outward assembly of growth units. Inward growth appears to be originated from myofibroblasts derived from small portal vessel around intravascular entrapped eggs, while outward growth arises from hepatic stellate cells. The auto-assembly of the growth units defines the three-dimensional scaffold of the schistosome granulomas. The granuloma surface irregularity and its border presented fractal dimension equal to 1.58. Second, it is internally regulated by intricate networks of immuneneuroendocrine stimuli orchestrated by leptin and leptin receptors, substance P and Vasoactive intestinal peptide. Third, it can reach the population of +/- 40,000 cells and presents an autopoietic component evidenced by internal proliferation (Ki-67+ Cells), and by expression of c-Kit+ Cells, leptin and leptin receptor (Ob-R), granulocyte-colony stimulating factor (G-CSF-R), and erythropoietin (Epo-R) receptors. Fourth, the granulomas cells are intimately connected by pan-cadherins, occludin and connexin-43, building a state of closing (granuloma closure). In conclusion, the granuloma is characterized by transitory stages in such a way that its organized structure emerges as a global property which is greater than the sum of actions of its individual cells and extracellular matrix components.
Asunto(s)
Granuloma/patología , Parasitosis Hepáticas/patología , Schistosoma mansoni/fisiología , Esquistosomiasis mansoni/patología , Animales , Modelos Animales de Enfermedad , Técnica del Anticuerpo Fluorescente Indirecta , Fractales , Granuloma/metabolismo , Granuloma/parasitología , Parasitosis Hepáticas/metabolismo , Parasitosis Hepáticas/parasitología , Ratones , Esquistosomiasis mansoni/metabolismo , Esquistosomiasis mansoni/parasitología , Coloración y Etiquetado , Factores de TiempoRESUMEN
We have previously showed that Schistosoma mansoni ATP-diphosphohydrolase and Solanum tuberosum potato apyrase share epitopes and the vegetable protein has immunostimulatory properties. Here, it was verified the in situ cross-immunoreactivity between mice NTPDases and anti-potato apyrase antibodies produced in rabbits, using confocal microscopy. Liver samples were taken from Swiss Webster mouse 8 weeks after infection with S. mansoni cercariae, and anti-potato apyrase and TRITC-conjugated anti-rabbit IgG antibody were tested on cryostat sections. The results showed that S. mansoni egg ATP diphosphohydrolase isoforms, developed by anti-potato apyrase, are expressed in miracidial and egg structures, and not in granulomatous cells and hepatic structures (hepatocytes, bile ducts, and blood vessels). Therefore, purified potato apyrase when inoculated in rabbit generates polyclonal sera containing anti-apyrase antibodies that are capable of recognizing specifically S. mansoni ATP diphosphohydrolase epitopes, but not proteins from mammalian tissues, suggesting that autoantibodies are not induced during potato apyrase immunization. A phylogenetic tree obtained for the NTPDase family showed that potato apyrase had lower homology with mammalian NTPDases 1-4, 7, and 8. Further analysis of potato apyrase epitopes could implement their potential use in schistosomiasis experimental models.
Asunto(s)
Adenosina Trifosfatasas/inmunología , Apirasa/inmunología , Schistosoma mansoni/enzimología , Esquistosomiasis mansoni/inmunología , Solanum tuberosum/enzimología , Adenosina Trifosfatasas/metabolismo , Secuencia de Aminoácidos , Animales , Anticuerpos Antihelmínticos/inmunología , Apirasa/metabolismo , Reacciones Cruzadas , Modelos Animales de Enfermedad , Masculino , Ratones , Microscopía Confocal , Datos de Secuencia Molecular , Conejos , Schistosoma mansoni/inmunología , Schistosoma mansoni/metabolismoRESUMEN
The fact that the Schistosoma mansoni egg has two ATP diphosphohydrolase (EC 3.6.1.5) isoforms with different net charges and an identical molecular weight of 63,000, identified by non-denaturing polyacrylamide gel electrophoresis and immunological cross-reactivity with potato apyrase antibodies, is shown. In soluble egg antigen (SEA), only the isoform with the lower net negative charge was detected and seemed to be the predominant species in this preparation. By confocal fluorescence microscopy, using anti-potato apyrase antibodies, the S. mansoni egg ATP diphosphohydrolase was detected on the external surface of miracidium and in von Lichtenberg's envelope. Intense fluorescence was also seen in the outer side of the egg-shell, entrapped by the surface microspines, suggesting that a soluble isoform is secreted. ATP diphosphohydrolase antigenicity was tested using the vegetable protein as antigen. The purified potato apyrase was recognized in Western blots by antibodies present in sera from experimentally S. mansoni-infected mice. In addition, high levels of IgG anti-ATP diphosphohydrolase antibodies were detected by ELISA in the same sera. This work represents the first demonstration of antigenic properties of S. mansoni ATP diphosphohydrolase and immunological cross-reactivity between potato apyrase and sera from infected individuals.
Asunto(s)
Antígenos Helmínticos/química , Apirasa/química , Schistosoma mansoni/enzimología , Animales , Antígenos Helmínticos/aislamiento & purificación , Antígenos Helmínticos/metabolismo , Apirasa/inmunología , Apirasa/aislamiento & purificación , Apirasa/metabolismo , Western Blotting , Electroforesis en Gel de Poliacrilamida , Inmunohistoquímica , Isoenzimas , Hígado/parasitología , Ratones , Microscopía Fluorescente , Peso Molecular , Schistosoma mansoni/inmunología , Schistosoma mansoni/metabolismoRESUMEN
Human abdominal angiostrongyliasis is a severe eosinophilic disease caused by Angiostrongylus costaricensis. Previous studies have demonstrated that wild rodents are critically involved as definitive hosts to this nematode in nature. In this study, we have evaluated the susceptibility of Wistar rats (Rattus norvegicus) to A. costaricensis infection. Kinetics of parasitological and pathological changes, including the number of adult worms recovered from mesenteric arteries, and of IgE, mast cell and eosinophil levels in several compartments have been assessed. The oral inoculation of third-stage larvae (L3) into adult Wistar rats led to a marked accumulation of worms in the branches of the mesenteric arteries 25 and 50 days post-inoculation. Intense bone marrow eosinophilia ranging from 7 to 50 days was accompanied by marked accumulation of eosinophils in the blood, peritoneal and bronchoalveolar spaces. Eosinophilic periarteritis, oedema and granuloma in the intestinal and lung tissues were also histologically evident. Total serum IgE and specific anti-parasite IgE peaked at 25 days post-infection, as measured by ELISA and by the passive cutaneous anaphylaxis test, respectively. At that time point, there was a drastic reduction in the number of intact mast cells in the peritoneal effluent. These findings indicate that Wistar rats are permissive to A. costaricensis infection. IgE-mast cell activation and massive tissue eosinophil infiltration are marked features in the process and are likely to play a crucial role in the immune-response evoked by this parasite.
Asunto(s)
Angiostrongylus/inmunología , Eosinófilos/patología , Inmunoglobulina E/inmunología , Mastocitos/patología , Infecciones por Strongylida/inmunología , Animales , Ensayo de Inmunoadsorción Enzimática , Cinética , Cavidad Peritoneal , Arteria Pulmonar/parasitología , Ratas , Ratas Wistar , Infecciones por Strongylida/patologíaRESUMEN
We previously reported that portal veins from mice infected with male Schistosoma mansoni exhibited an increased reactivity to 5-hydroxytryptamine (5-HT). Here, we extended our observations to mice infected by both male and female worms and we further investigated another constrictor agent and the mechanism(s) responsible for the enhanced maximal contraction ( E(max)). Bisexual infection increased the E(max) of 5-HT (from 0.66+/-0.06 mN.s to 1.56+/-0.38 mN.s), in a similar way to the unisexual (male) infection. Infection with male worms increased portal vein reactivity to acetylcholine, as revealed by a higher E(max) (1.03+/-0.2 mN.s) in relation to non-infected control animals ( E(max)= 0.54+/-0.08 mN.s). Sarcoplasmic/endoplasmic reticulum Ca(2+)-ATPase (SERCA) inhibition with 100 nM thapsigargin reduced the E(max) of 5-HT by 35% in both tissues, discharging a deficiency of SERCA pump in infected animals. In contrast, the number of voltage-dependent Ca(2+) channels (L-type) was higher in portal veins from infected than non-infected control mice. Inhibition of Ca(2+)-activated chloride channels (Cl(Ca)) with 10 micro M niflumic acid reduced the E(max) of 5-HT in portal veins more from infected than non-infected animals (remaining tension = 60.9+/-2.2% and 70.4+/-2.3%, respectively). Histopathological analysis revealed an increased content of collagen and elastin in portal veins from male S. mansoni-infected mice, compatible with an increased intraluminal pressure. In conclusion, male S. mansoni altered portal vein physiology, increasing the E(max) of two vasoconstrictors, possibly by increasing membrane depolarisation through a more effective opening of Cl(Ca) channels, with calcium entering through L-type Ca(2+) channels.
Asunto(s)
Vena Porta/fisiopatología , Schistosoma mansoni/patogenicidad , Esquistosomiasis mansoni/fisiopatología , Vasoconstricción , Acetilcolina/farmacología , Animales , Calcio/metabolismo , Relación Dosis-Respuesta a Droga , Femenino , Histocitoquímica , Masculino , Ratones , Vena Porta/citología , Vena Porta/patología , Retículo Sarcoplasmático/efectos de los fármacos , Retículo Sarcoplasmático/metabolismo , Esquistosomiasis mansoni/metabolismo , Esquistosomiasis mansoni/parasitología , Esquistosomiasis mansoni/patología , Serotonina/farmacología , Factores Sexuales , CaracolesRESUMEN
The objective of the present study was to investigate whether the injection of a tolerated protein (indirect effects) affects the formation of granulomas around Schistosoma mansoni eggs trapped in the lungs after intravenous (iv) injection into normal (noninfected) C57BL/6 mice (6 animals per group). To induce oral tolerance to chicken egg ovalbumin a 1/5 dilution of egg white in water was offered ad libitum in a drinking bottle for 3 days. Control mice received water. After 7 days, control and experimental animals were injected iv with 2,000 S. mansoni eggs through a tail vein. In some mice of both groups the iv injection of eggs was immediately followed by intraperitoneal (ip) immunization with 10 micro g of dinitrophenylated conjugates of ovalbumin (DNP-Ova) emulsified in complete Freund's adjuvant (CFA) or only CFA; 18 days later, mice were bled and killed by ether inhalation. The lungs were fixed in formalin and embedded in paraffin. Serial sections of 5 m were stained with Giemsa, Gomori's silver reticulin and Sirius red (pH 10.2). Granuloma diameters were measured in histological sections previously stained with Gomori's reticulin. Anti-DNP and anti-soluble egg antigen (SEA) antibodies were analyzed by ELISA. In mice orally tolerant to ovalbumin the concomitant ip injection of DNP-Ova resulted in significantly lower anti-SEA antibodies (ELISA*: 1395 +/- 352 in non-tolerant and 462 +/- 146 in tolerant mice) and affected granuloma formation around eggs, significantly decreasing granuloma size (area: 22,260 +/- 2478 to 12,993 +/- 3242 m ). Active mechanisms triggered by injection of tolerated antigen (ovalbumin) reduce granuloma formation.
Asunto(s)
Antígenos Helmínticos/inmunología , Dinitrofenoles/inmunología , Granuloma/parasitología , Haptenos/inmunología , Enfermedades Pulmonares Parasitarias/inmunología , Ovalbúmina/administración & dosificación , Ovalbúmina/inmunología , Schistosoma mansoni/inmunología , Administración Oral , Animales , Dinitrofenoles/administración & dosificación , Ensayo de Inmunoadsorción Enzimática , Granuloma/inmunología , Granuloma/patología , Haptenos/administración & dosificación , Tolerancia Inmunológica , Enfermedades Pulmonares Parasitarias/patología , Ratones , Ratones Endogámicos BALB CRESUMEN
The objective of the present study was to investigate whether the injection of a tolerated protein (indirect effects) affects the formation of granulomas around Schistosoma mansoni eggs trapped in the lungs after intravenous (iv) injection into normal (noninfected) C57BL/6 mice (6 animals per group). To induce oral tolerance to chicken egg ovalbumin a 1/5 dilution of egg white in water was offered ad libitum in a drinking bottle for 3 days. Control mice received water. After 7 days, control and experimental animals were injected iv with 2,000 S. mansoni eggs through a tail vein. In some mice of both groups the iv injection of eggs was immediately followed by intraperitoneal (ip) immunization with 10 æg of dinitrophenylated conjugates of ovalbumin (DNP-Ova) emulsified in complete Freund's adjuvant (CFA) or only CFA; 18 days later, mice were bled and killed by ether inhalation. The lungs were fixed in formalin and embedded in paraffin. Serial sections of 5 æm were stained with Giemsa, Gomori's silver reticulin and Sirius red (pH 10.2). Granuloma diameters were measured in histological sections previously stained with Gomori's reticulin. Anti-DNP and anti-soluble egg antigen (SEA) antibodies were analyzed by ELISA. In mice orally tolerant to ovalbumin the concomitant ip injection of DNP-Ova resulted in significantly lower anti-SEA antibodies (ELISA*: 1395 ± 352 in non-tolerant and 462 ± 146 in tolerant mice) and affected granuloma formation around eggs, significantly decreasing granuloma size (area: 22,260 ± 2478 to 12,993 ± 3242 æmý). Active mechanisms triggered by injection of tolerated antigen (ovalbumin) reduce granuloma formation
Asunto(s)
Animales , Ratones , Antígenos Helmínticos , Enfermedades Pulmonares Parasitarias , Ovalbúmina , Schistosoma mansoni , Administración Oral , Ensayo de Inmunoadsorción Enzimática , Tolerancia Inmunológica , Enfermedades Pulmonares Parasitarias , Ratones Endogámicos BALB C , OvalbúminaRESUMEN
Malnutrition hampers the course of schistosomiasis mansoni infection just as normal growth of adult worms. A comparative morphometric study on adult specimens (male and female) recovered from undernourished (fed with a low protein diet - regional basic diet) and nourished (rodent commercial laboratory food, NUVILAB) white mice was performed. Tomographic images and morphometric analysis of the oral and ventral suckers, reproductive system and tegument were obtained by means of confocal laser scanning microscopy. Undernourished male specimens presented smaller morphometric values (length and width) of the reproductive system (first, third and last testicular lobes) and thickness of the tegument than controls. Besides that, it was demonstrated that the dorsal surface of the male worms bears large tubercles unevenly distributed, but kept grouped and flat. At the subtegumental region, vacuolated areas were detected. It was concluded that the inadequate nutritional status of the vertebrate host has a negative influence mainly in the reproductive system and topographical somatic development of male adult Schistosoma mansoni, inducing some alterations on the structure of the parasite.
Asunto(s)
Estado Nutricional , Schistosoma mansoni/ultraestructura , Animales , Femenino , Interacciones Huésped-Parásitos , Masculino , Ratones , Microscopía Confocal , Trastornos Nutricionales/parasitología , Schistosoma mansoni/crecimiento & desarrollo , Schistosoma mansoni/aislamiento & purificación , Esquistosomiasis mansoni/parasitologíaRESUMEN
Mast cells and eosinophils actively participate in tissue repair and are prominent components of Schistosoma mansoni granulomas. Since pentoxifillyne (PTX) is an immunomodulatory and antifibrotic substance, we aimed to characterize, by morphological techniques, the effect of this drug on fibrosis developed inside murine hepatic schistosomal granulomatous reaction, beyond the quantification of eosinophil and mast cell populations. The drug (1 mg/100 g animal weight) was administrated from 35 to 90 days post-infection, when the animals were killed. The intragranulomatous interstitial collagen network was analyzed by confocal laser scanning microscopy, the number of eosinophils and mast cells was quantified and the results were validated by t-student test. Treatment did not interfere on the granuloma evolution but caused a significant decrease in the total and involutive number of hepatic granulomas (p = 0.01 and 0.001, respectively), and in the intragranulomatous accumulation of eosinophils (p = 0.0001). Otherwise, the number of mast cells was not significantly altered (p = 0.9); however, it was positively correlated with the number of granulomatous structures (r = 0.955). In conclusion, PTX does not affect development and collagen deposition in S. mansoni murine granuloma, but decreases the intragranulomatous eosinophil accumulation possibly due to its immunomodulatory capability, interfering in cellular recruitment and/or differentiation.
Asunto(s)
Eosinófilos/efectos de los fármacos , Matriz Extracelular/efectos de los fármacos , Granuloma/patología , Parasitosis Hepáticas/patología , Pentoxifilina/farmacología , Inhibidores de Fosfodiesterasa/farmacología , Esquistosomiasis mansoni/complicaciones , Animales , Colágeno/efectos de los fármacos , Granuloma/tratamiento farmacológico , Cirrosis Hepática/patología , Parasitosis Hepáticas/tratamiento farmacológico , Masculino , Mastocitos/efectos de los fármacos , Ratones , Pentoxifilina/uso terapéutico , Inhibidores de Fosfodiesterasa/uso terapéuticoRESUMEN
This study examined the dynamics of colonization of Trypanosoma cruzi in the scent glands of the opossum Didelphis marsupialis following direct inoculation with 10(5) epimastigotes of isolate G-49 (an opossum-derived strain). One, three, and five days, 1 month, and 1 year after inoculation, scent glands were fixed for analysis using brightfield and electron microscopies. One day after inoculation the parasites, mainly as epimastigotes, were randomly distributed into the lumen. From the third day on, the parasites still in the form of epimastigotes tended to concentrate closer to the epithelium. The flagellates reached the definitive distribution pattern on the fifth day, when they formed huge clusters deep into the foveae. In samples collected 1 month and 1 year after inoculation, the ratio of epimastigotes:trypomastigotes was 1:1, with epimastigotes predominating near the epithelium and trypomastigotes far from it. Our observations suggest that T. cruzi grows continuously in the scent glands and does not depend on adhesion to promote metacyclogenesis. Metacyclogenesis far from the epithelium seems to be an important selective advantage to both host and parasite, since it assures the elimination of the infective forms of the parasite when the host expels the glands' contents, which occurs in frightening situations or at times of stress. The morphological characteristics of infected and noninfected scent glands using transmission and scanning electron microscopies were also described.
Asunto(s)
Enfermedad de Chagas/veterinaria , Zarigüeyas/parasitología , Glándulas Odoríferas/parasitología , Trypanosoma cruzi/crecimiento & desarrollo , Animales , Enfermedad de Chagas/parasitología , Enfermedad de Chagas/transmisión , Reservorios de Enfermedades/veterinaria , Vectores de Enfermedades , Femenino , Microscopía Electrónica/veterinaria , Microscopía Electrónica de Rastreo/veterinaria , Glándulas Odoríferas/ultraestructura , Trypanosoma cruzi/ultraestructuraRESUMEN
This work is an investigation on the microvasculature of the cutaneous infiltrates of leprosy with the immunohistochemical staining of endothelial cells in cutaneous biopsies. Anti-Factor VIII-related antigen antibody (anti-FVIII-ra) and Ulex Europaeus-1 lectin (UEA-1) binding were utilized as endothelial cell markers. Thirty-nine patients grouped according to the Ridley-Jopling classification (14 borderline tuberculoid, 18 borderline lepromatous, 6 lepromatous, and 1 indeterminate leprosy) were selected for this study. Two microvascular architectural patterns could be clearly distinguished: lepromatous lesions presented a dense and tortuous mesh of microvessels among the Mycobacterium leprae-glutted macrophages; whereas the microvessels in the tuberculoid lesions were restricted to the periphery of the granulomas and were not seen among the central epithelioid cells. We were able to distinguish three basic morphological kinds of infiltrate distribution related to the microvessels: micronodules, cords and macronodules. Intensifications of the FVIII-ra immunoreactivity and UEA-1 binding capacity were observed in the endothelial cells of microvessels involved by the inflammatory infiltrate. A distinct cytokine expression profile at the leprosy poles and the role of mast cells in angiogenesis were speculated as factors contributing to these distinct patterns. Growth of the lesion and systemic dissemination of M. leprae in the bipolar spectrum of leprosy may hypothetically be influenced by the vascular-infiltrate relationship. The detection of angiogenesis in the cutaneous lesions of leprosy may bring about alternate and/or additional strategies for leprosy treatment.
Asunto(s)
Lepra Lepromatosa/patología , Lepra Tuberculoide/patología , Piel/irrigación sanguínea , Biopsia , Endotelio Vascular/patología , Granuloma , Humanos , Inmunohistoquímica , Piel/patología , Factor de von Willebrand/aislamiento & purificaciónRESUMEN
Plasmodium berghei ANKA infection in CBA/J mice leads to the development of cerebral malaria (CM) that kills 80-90% of the animals in 6-9 days. This model has been used to study the pathogenesis of CM, which is a major cause of morbidity and mortality in Plasmodium falciparum-infected individuals. The role of cytokines in the induction of CM in the murine model has been well documented, but most studies have been restricted to the peak of neurological manifestations. Here we used a sequential approach to compare mice that developed CM with those that developed no cerebral pathology. Animals were examined for systemic histopathological changes and plasma Tumor Necrosis Factor-alpha (TNF) levels. The objectives were (a) to further determine the importance of factors commonly associated with murine CM-such as elevated levels of TNF and the presence of hemorrhage and vascular plugging-by comparing mice at different stages of infection and/or with different outcomes following infection and (b) to examine the importance of systemic changes-course of parasitemia and histopathological alterations in brain, liver, and lungs-in the development of CM. The data suggest that (a) the clinical manifestation of CM appears to be associated with a wave of merozoite release on days 6-7, (b) murine CM does not present reliable histopathological indicators, (c) there is no topographic association between the occurrence of intravascular plugging and the hemorrhagic foci, (d) monocyte-monocyte and monocyte-endothelial cell adherence were the most expressive histopathological events and were not restricted to brain vessels, (e) blood levels of TNF are not indicative of the local tissue reaction, (f) adhesiveness of monocyte/endothelial cells fluctuate during infection and is dissociated from the lymphocyte homing to the liver, and (g) pulmonary megakaryocytosis (megakaryopoiesis?) is a late event in the lungs.
Asunto(s)
Malaria Cerebral/inmunología , Plasmodium berghei , Factor de Necrosis Tumoral alfa/análisis , Animales , Encéfalo/patología , Modelos Animales de Enfermedad , Femenino , Hígado/patología , Pulmón/patología , Malaria Cerebral/patología , Ratones , Ratones Endogámicos CBARESUMEN
In noninfected rats, challenge with allergen following local IgE sensitization induced a pleurisy marked by intense protein exudation that plateaued from 30 min to 4 h after challenge, reducing thereafter. Infection of rats with Angiostrongylus costaricensis induced a 5-fold increase in blood eosinophil numbers by 25 days postinfection, whereas the numbers of eosinophils in the pleural cavity ranged from normal to a weak increase. In infected rats, identically sensitized, challenge with Ag induced a much shorter duration of pleural edema with complete resolution by 4 h, but no change in the early edema response. In parallel, infection increased the number of eosinophils recovered from the pleural cavity at 4 h, but not at 30 min, following allergen challenge. Pretreatment with IL-5 (100 IU/kg, i.v.) also increased eosinophil numbers in blood and, after allergen challenge, shortened the duration of the pleural edema and increased pleural eosinophil numbers. There were increases in the levels of both PGE2 and lipoxin A4 (LXA4) in pleural exudate. Selective cyclooxygenase (COX)-2 inhibitors, NS-398, meloxicam, and SC-236, did not alter pleural eosinophilia, but reversed the curtailment of the edema in either infected or IL-5-pretreated rats. Pretreatment of noninfected animals with the PGE analogue, misoprostol, or two stable LXA4 analogues did not alter the magnitude of pleural exudation response, but clearly shortened its duration. These results indicate that the early resolution of allergic pleural edema observed during A. costaricensis infection coincided with a selective local eosinophilia and seemed to be mediated by COX-2-derived PGE2 and LXA4.
Asunto(s)
Angiostrongylus/inmunología , Dinoprostona/fisiología , Edema/terapia , Eosinofilia/enzimología , Ácidos Hidroxieicosatetraenoicos/fisiología , Hipersensibilidad/terapia , Isoenzimas/metabolismo , Lipoxinas , Prostaglandina-Endoperóxido Sintasas/metabolismo , Infecciones por Strongylida/enzimología , Administración Oral , Animales , Antiinflamatorios no Esteroideos/metabolismo , Antiinflamatorios no Esteroideos/farmacología , Antígenos Helmínticos/administración & dosificación , Corticosterona/sangre , Ciclooxigenasa 2 , Inhibidores de la Ciclooxigenasa 2 , Inhibidores de la Ciclooxigenasa/administración & dosificación , Dinoprostona/metabolismo , Edema/enzimología , Edema/patología , Edema/fisiopatología , Eosinofilia/patología , Eosinofilia/fisiopatología , Exudados y Transudados/efectos de los fármacos , Exudados y Transudados/enzimología , Femenino , Ácidos Hidroxieicosatetraenoicos/metabolismo , Hipersensibilidad/enzimología , Hipersensibilidad/patología , Hipersensibilidad/fisiopatología , Inyecciones Intraperitoneales , Inyecciones Intravenosas , Interleucina-5/administración & dosificación , Isoenzimas/farmacología , Cinética , Leucotrieno C4/metabolismo , Masculino , Misoprostol/administración & dosificación , Derrame Pleural/enzimología , Derrame Pleural/metabolismo , Derrame Pleural/patología , Derrame Pleural/prevención & control , Pleuresia/enzimología , Pleuresia/patología , Pleuresia/fisiopatología , Prostaglandina-Endoperóxido Sintasas/farmacología , Ratas , Ratas Wistar , Infecciones por Strongylida/patología , Infecciones por Strongylida/fisiopatologíaRESUMEN
Most of our knowledge concerning the virulence determinants of pathogenic fungi comes from the infected host, mainly from animal models and more recently from in vitro studies with cell cultures. The fungi usually present intra- and/or extracellular host-parasite interfaces, with the parasitism phenomenon dependent on complementary surface molecules. Among living organisms, this has been characterized as a cohabitation event, where the fungus is able to recognize specific host tissues acting as an attractant, creating stable conditions for its survival. Several fungi pathogenic for humans and animals have evolved special strategies to deliver elements to their cellular targets that may be relevant to their pathogenicity. Most of these pathogens express surface factors that mediate binding to host cells either directly or indirectly, in the latter case binding to host adhesion components such as extracellular matrix (ECM) proteins, which act as 'interlinking' molecules. The entry of the pathogen into the host cell is initiated by fungal adherence to the cell surface, which generates an uptake signal that may induce its cytoplasmic internalization. Once this is accomplished, some fungi are able to alter the host cytoskeletal architecture, as manifested by a rearrangement of microtubule and microfilament proteins, and this can also induce epithelial host cells to become apoptotic. It is possible that fungal pathogens induce modulation of different host cell pathways in order to evade host defences and to foster their own proliferation. For a number of pathogens, the ability to bind ECM glycoproteins, the capability of internalization and the induction of apoptosis are considered important factors in virulence. Furthermore, specific recognition between fungal parasites and their host cell targets may be mediated by the interaction of carbohydrate-binding proteins, e.g., lectins on the surface of one type of cell, probably a parasite, that combine with complementary sugars on the surface of host-cell. These interactions supply precise models to study putative adhesins and receptor-containing molecules in the context of the fungus-host interface. The recognition of the host molecules by fungi such as Aspergillus fumigatus, Paracoccidioides brasiliensis and Histoplasma capsulatum, and their molecular mechanisms of adhesion and invasion, are reviewed in this paper.
Asunto(s)
Aspergillus fumigatus/patogenicidad , Histoplasma/patogenicidad , Paracoccidioides/patogenicidad , Animales , Aspergillus fumigatus/fisiología , Adhesión Celular , Línea Celular , Histoplasma/fisiología , Humanos , Micosis/microbiología , Paracoccidioides/fisiología , VirulenciaRESUMEN
The intermediate hosts of Angiostrongylus costaricensis are terrestrian molluscs, mostly of the family Veronicellidae. The present work aimed at clarifying more accurately the sites of penetration and the migratory routes of A. costaricensis in the tissue slugs and at verifying the pattern of the perilarval reaction at different times of infection. Slugs were individually infected with 5,000 L1, and killed from 30 min to 30 days after infection. From 30 min up to 2 hr after infection, L1 were found within the lumen of different segments of the digestive tube having their number diminished in more advanced times after exposition until complete disappearance. After 30 min of exposition, percutaneous infection occurred, simultaneously to oral infection. Perilarval reaction was observed from 2 hr of infection around larvae in fibromuscular layer, appearing later (after 6 hr) around larvae located in the viscera. A pre-granulomatous reaction was characterized by gradative concentration of amebocytes around larvae, evolving two well-organized granulomas. In this work we confirmed the simultaneous occurrence of oral and percutaneous infections. Perilarval reaction, when very well developed, defined typical granulomatous structure, including epithelioid cell transformation. The infection also caused a systemic mobilization of amebocytes and provoked amebocyte-endothelium interactions.
Asunto(s)
Angiostrongylus/fisiología , Interacciones Huésped-Parásitos , Moluscos/parasitología , Infecciones por Strongylida/parasitología , Angiostrongylus/química , AnimalesRESUMEN
The collagen structure of isolated and in situ liver granuloma from Swiss Webster mice infected with Schistosoma mansoni was sequentially and three-dimensionally analyzed during different times of infection (early acute, acute, transitional acute-chronic, and chronic phases) by laser scanning confocal microscopy and electron scanning variable vacuum microscopy. The initial granuloma structure is characterized by vascular collagen residues and by anchorage points (or fiber radiation centers), from where collagenous fibers are angularly shed and self-assembled. During the exudative-productive stage, the self-assembly of these fibers minimizes energy and mass through continuous tension and focal compression. The curvature or angles between collagen fibers probably depends on the fibroblastic or myofibroblastic organization of stress fibers. Gradually, the loose unstable lattice of the exudative-productive stage transforms into a highly packed and stable architecture as a result of progressive compactness. The three-dimensional architecture of granulomas provides increased tissue integrity, efficient distribution of soluble compounds and a haptotactic background to the cells.
Asunto(s)
Colágeno/análisis , Granuloma/patología , Parasitosis Hepáticas/patología , Esquistosomiasis mansoni/patología , Animales , Colágeno/ultraestructura , Matriz Extracelular/química , Matriz Extracelular/ultraestructura , Fibroblastos , Ratones , Microscopía ConfocalRESUMEN
The collagen structure of isolated and in situ liver granuloma from Swiss Webster mice infected with Schistosoma mansoni was sequentially and three-dimensionally analyzed during different times of infection (early acute, acute, transitional acute-chronic, and chronic phases) by laser scanning confocal microscopy and electron scanning variable vacuum microscopy. The initial granuloma structure is characterized by vascular collagen residues and by anchorage points (or fiber radiation centers), from where collagenous fibers are angularly shed and self-assembled. During the exudative-productive stage, the self-assembly of these fibers minimizes energy and mass through continuous tension and focal compression. The curvature or angles between collagen fibers probably depends on the fibroblastic or myofibroblastic organization of stress fibers. Gradually, the loose unstable lattice of the exudative-productive stage transforms into a highly packed and stable architecture as a result of progressive compactness. The three-dimensional architecture of granulomas provides increased tissue integrity, efficient distribution of soluble compounds and a haptotactic background to the cells
Asunto(s)
Animales , Ratones , Colágeno/análisis , Granuloma/patología , Hepatopatías/patología , Esquistosomiasis mansoni/patología , Colágeno/ultraestructura , Matriz Extracelular/química , Matriz Extracelular/ultraestructura , Fibroblastos , Microscopía ConfocalRESUMEN
The development of genetically modified vectors refractory to parasites is seen as a promising strategy in the future control of endemic diseases such as malaria. Nevertheless, knowledge of mosquito embryogenesis, a pre-requisite to the establishment of transgenic individuals, has been presently neglected. We have here studied the eggs from two neotropical malaria vectors. Eggs from Anopheles (Nyssorhynchus) albitarsis and Anopheles (Nyssorhynchus) aquasalis were analyzed by laser scanning microscopy and scanning electron microscopy and compared to those of Drosophila melanogaster. We verified basic conflicting data such as mosquito egg polarity and ultrastructure of eggshell layers. A 180 degrees rotation movement of the mosquito embryo along its longitudinal axis, a phenomenon not conserved among all Diptera, was confirmed. This early event is not taken into account by several present groups, leading to a non-consensual assignment of eggshell dorsal and ventral poles. Since embryo and egg polarities, defined during oogenesis, are the same, we propose to consider the flattened egg side as the dorsal one. The structure of Anopheles eggshell was also examined. Embryos are covered by a smooth endochorion or inner chorion layer. Outside this coat lies the compound exochorion or outer chorion layer, assembled by a thin basal lamellar layer and external tubercles. The terminology related to eggshell layers is discussed.