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Objective To establish an experimental model of acute cerebral schistosomiasis japonica and explore the MRI manifestations of acute cerebral schistosomiasis. Methods Rabbits were divided into 3 groups with 10 rabbits in each group. The rabbits in the experimental group were directly injected with suspension fluid of Schistosoma japonicum eggs(0.9 mg,1 ml) by the cranial drilling method,those in the negative control group were given saline(1 ml)by the same method above-men-tioned,and those in the blank control group were not given any treatment. Antibiotic was given to the first two groups after the op-eration. The clinical manifestations of the 3 groups were observed,and the magnetic resonance imaging(MRI)was performed in 30 days post-operation,and then the brain tissues were taken for pathological examinations. Results All the rabbits in the ex-perimental group exhibited inappetence,various neurological symptoms including hemiplegia,and weight loss after the opera-tion;while those in the negative control group showed inappetence in 3 days after the operation,and 1 week later,the symptom disappeared;there were no adverse reactions in the blank control group. MRI of the experimental group showed nodular or patchy enhancement on T1WI enhancement,brain edema,abnormal ventricular dilatation,and needle augmentation. SWI dis-played hypointense in the abnormal enhanced nodules and flaky hypointense on the operation brain. In the negative control group,2 rabbits showed abnormal enhancement of the needle canal,and 1 showed mild dilatation of the ventricle. The blank control group showed normal manifestations. The pathological examinations showed abnormal appearances in 10 rabbits of the ex-perimental group,including 6 with S. japonicum egg granuloma nodules,nonspecific granuloma nodules coexisted with perivas-cular inflammation;no granuloma nodules were found in the negative control group,but 2 rabbits showed vascular inflamma-tion;the blank control group showed the normal brain tissue. Conclusions An experimental model of acute cerebral schistoso-miasis is successfully established in rabbits by intracranial injection of schistosome eggs. The MRI examination combined with the clinical manifestations can improve the accuracy of early diagnosis of cerebral schistosomiasis.
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To determine immune responses and immunopathology in ICOSL knockout (ICOSL KO) mice infected with Schistosoma japonicum,ICOSL- KO mice and wild-type C57BL/6J mice were used as experimental models for Schistosoma japonicum infection.The splenic lymphocytes were isolated from the mice the day before infection (0 week) as well as 4,7,12,16 and 20 weeks post infection,and stimulated with SEA for 72 hours in culture.The concentrations of Th1 cytokines (IFN-γand IL- 12) and Th2 cytokines (IL- 4,IL-10 and IL-13) in the culture supernatants were measured by sandwich ELISA.The levels of SEA-specific IgG antibody and its subtypes (IgG1 and IgG2a) were measured in mouse sera by ELISA.Pathological changes of hepatic granuloma in mice were determined by hematoxylin and eosin (H&E) staining.After the infection,the levels of Th1 cytokines,IFN- γ and IL 12,in ICOSL- KO mice were higher than those in wild-type C57BL/6J mice.However,the levels of Th2 cytokines (IL- 4,IL- 10 and IL- 13) were significantly decreased in ICOSL-KO mice compared to those in wild-type C57BL/6J mice.The levels of SEA-specific IgG antibody and its subtypes (IgG1 and IgG2a) in the sera of ICOSL- KO mice were also significantly lower than those of wild -type C57BL/6J mice.Moreover,the Th2 differentiation index was lower in ICOSL- KO mice than in wild-type C57BL/6J mice at 4,7,12,16 and 20 weeks post-infection.Similarly,the ratio of IgG1/IgG2a in ICOSL-KO mice was significantly lower than that in wild- type C57BL/6J mice at 7,12 and 16 weeks post- infection.Furthermore,throughout the course of disease progression,the volume of hepatic egg granuloma in ICOSL- KO mice was significantly smaller than that in wild-type C57BL/6J mice.In conclusions,there is a substantially down-regulated Th2 immune response in ICOSL- KO mice infected with Schistosoma japonicum,thus results in an attenuated hepatic lesion caused by egg granulomas.The findings indicate that the ICOS ICOSL co-stimulatory pathway plays an important role in the hepatic egg granuloma formation of schistosomiasis.
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This study aims to evaluate the egg-granuloma system in hepatic tissues using lectin histochemistry in experimental Schistosomiasis. Eight Swiss mice were infected with a local strain of Schistosoma mansoni, being submitted forty days later to a perfusion after which slices of liver were prepared. The tissue samples were incubated with the following peroxidase conjugated lectins: Peanut agglutinin (PNA), Wheat Germ agglutinin (WGA), and Concanavalin A (Con A). All lectins recognized the glycoconjugates in the adult worm tegument. In the hepatic tissue, WGA presented the highest staining followed by PNA and Con A. The PNA presented the most intense staining of the egg-granuloma system while WGA stained the hepatic sinusoid cells and Con A bound preferentially the fibrosis rings of granuloma and the surrounding hepatic parenquima. WGA and PNA indicated the presence of residues of N-acetyl-glucosamine and galactose in the surface of Schistosoma mansoni eggs in the hepatic granulomas. In conclusion, using PNA, Con A and WGA our study presented different aspects of the egg-granuloma and Tegument of Schistosoma mansoni as well as indicated differences in the peri-ovular granulomas indicating alterations in the cellular mechanism of expression of surface carbohydrates during progression of the Schistosomiasis.
El objetivo del estudio fue evaluar el sistema de los huevos de los granulomas en los tejidos hepáticos, utilizando histoquímica de lectinas esquistosomiasis. Ocho ratones suizos experimentales fueron infectados con una cepa local de Schistosoma mansoni y luego a los cuarenta días fueron sometidos a la perfusión y se prepararon cortes de hígado. Las muestras de los tejidos fueron incubadas con las siguientes peroxidasas lectinas conjugadas: aglutinina de maní (PNA), aglutinina de germenn de trigo (WGA), Concanavalin A (Con A). Todas las lectinas reconocieron las glicoconjugadas en el tegumento del gusano adulto. El tejido hepático con WGA presentó mayor coloración seguido de PNA y Con A. El PNA presentó la más intensa tinción de los huevos mientras el granuloma del sistema WGA tiñó las células hepáticas sinusoides y las Con A estuvieron siempre presentes en los anillos de la fibrosis y alrededor de los granulomas hepáticos del parénquima. WGA y PNA indicaron la presencia de residuos de N - acetil - glucosamina y galactosa en la superficie de los huevos de Schistosoma mansoni en los granulomas hepáticos de esquistosomiasis.
Subject(s)
Rats , Animals , Carbohydrates/analysis , Schistosomiasis mansoni/metabolism , Liver Diseases/metabolism , Liver Diseases/parasitology , Lectins/metabolism , Schistosoma mansoni/physiology , Disease Models, Animal , Schistosomiasis mansoni/chemically induced , Granuloma/metabolism , Granuloma/parasitology , Histocytochemistry , Ovum/physiologyABSTRACT
Ascariasis is probably the most common helminthic infestation of man, but it seldom causes severe illness. Pathologic conditions of Ascaris may be caused by adult worms, eggs or larvae. We describe a case of Ascaris egg granulomas that were found incidentally on the surface of the liver in a 75-year-old woman who had undergone a segmentectomy for an intrahepatic stone. Grossly, there were several yellowish calcific nodules of 0.4 cm in diameter on the lateral surface of the left lobe of the liver. Microscopically, the lesions were located in the hepatic capsule and consisted of fibrocalific nodules with many eggs. The eggs were round to oval, thick-shelled and measured 50~75x30~50 um. Most of the morphologically preserved eggs were fertilized eggs, but they had smooth shells without external protein coats. This case is of interest for the unusual location of the lesion, the presence of eggs without mammillation, and the association with the intrahepatic stone.
Subject(s)
Adult , Male , Female , HumansABSTRACT
Objective To further observe the morphologic changes of in vitro model of granulomatous responses around Schistosoma japonica eggs and determine the relationship of reaction inten-suy to infection stage of those schistosome-infected mice from which the splenocytes were obtained for in vitro experiments.Methods The spleen cells isolated from schistosome-infected mice were incubated and dry schistosome eggs were added into those cultures. These splenocytes responses surrounding eggs were observed and counted under the microscope.Results The splenocytes from infected mice showed obvious adherent responses to the dry eggs. Adhering cells were,at first,granulocytes,then replaced by the macrophages. Around the eggs proliferating lymphocytes,later fibroblasts, were found. The reaction index (RI) of the spleen cells from infected mice were significantly higher (P
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Objective To study the molecular mechanism of apoptosis of cells in egg granuloma induced by anti-idiotypic antibody NP30 of Schistosoma japonicum.Methods BALB/c mice were randomly divided into two groups. The mice of the experimental group were immunized by injecting NP30 intraperitoneally for three times, while the mice of control group were injected normal saline intraperitoneally. The mice were sacrificed respectively on the 39th, 49th, 64th, 108th, 112nd day after challenge with schistosome cercariae. The expressions of apoptosis-related gene Bax, Bcl-2, death receptor Fas, FasL (Fas ligand) and c-Fos were examined by the S-P method of immunohistochemistry,and Bax, mRNA and Fas mRNA investigated by the in-situ hybridization. Results The expressions of Bax, Fas, FasL and c-Fos were positive in granuloma cells of both groups. The expressions of Bax and FasL in experimental group were higher than those in control group (P
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Objective To study the expression of inducible nitric oxide synthase (iNOS) in livers of mice infected with Schistosoma japonicum. Methods The livers of NMRI mice infected with S. japonicum were collected on day 21, 28, 38, 45 post infection(p.i.), total RNA of livers were extracted and kinetics of the mRNA expression of iNOS were detected by RT-PCR, the protein expression of iNOS was then confirmed by Western blotting and the distribution of iNOS in the infected liver was determined by immunohistochemical methods. Results The mRNA expression of iNOS was not detectable in the uninfected liver, iNOS mRNA expression was detected on day 21 p.i, the expression increased on day 28 p.i and peaked on day 38 p.i, then decreased slightly on day 45 p.i. Western blotting showed an iNOS expression in the livers only on day 38, 45 p.i. IFA test showed that the expression of iNOS was maily distributed in the granuloma of the livers. Conclusion S. japonicum infection can induce the expression of iNOS in a time-dependent manner in the liver of the host,and eggs may be the main factor in inducing the expression.
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Objective To study the effects of the monoclonal anti idiotypic antibody NP30 active immunization on egg granuloma formation and hepatic fibrosis in Schistosoma japonicum infection. Methods ICR mice were actively immunized with NP30 100 ?g ?3 ip. every 10 days while the mice in control group were injected with SP2/0 ascites ip. simultaneously. After cercariae challenging,the mice were killed at the 4th, 8th,12th, 16th, 20th and 24th week, respectively.Mouse livers were removed and stained histochemically with VG and subjected to immunohistochemical assay of collagen type Ⅰ,Ⅲ and fibronectin(FN).The volume of egg granulomas and the content of collagen type Ⅰ,Ⅲ and FN were determined quantitatively by NYD 1000 Image Analysis System. Results The volume of egg granulomas in NP30 immunized group was much smaller than that of control group from the 12th week after cercariae challenge. The cellular components of egg granulomas in NP30 immunized group were significantly different from those of the control group,exhibiting two types of atypical egg granulomas were found.VG stain revealed that the average optical density of collagen in hepatic granulomas of experimental group was lower than that of control group.Immunohistochemical assay revealed that the contents of collagen type Ⅰ,Ⅲ and fibronectin in egg granulomas of experimental group were lower than those of control group. Conclusion NP30 vaccination may induce both cellular and humoral protective immunity to modulate egg granulomas and suppress liver fibrosis of schistosomiasis japonica.
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Fourteen-34 days after Schistosoma japonzcum-infected mice were injected intraperitoneally with immunoenhancing reagent,Chinese angelica root extract ("425"),the specific antibody (IgG) levels were significantly higher in Group Ⅲ (362.67?162.21-480.00?289.45) than the control group (66.67?39.68-245.33?101.56) in experiment one,and in Gronp Ⅱ(488.00?320.38-768.00?267.38) than the control group (256.00?189.07-394.67?141.06) in experiment two.The egg granulomatous formation were markedly diminished in the injected mice.The ratios of granulomas vs.eggs' mean diameters of Group in of mice were 5.24?1.04-3.95?0.77 and those of the control group were 6.59?1.19-5.29?0.94 in experiment one,those of the group Ⅱ were 3.75?0.71-4.15?0.73 and those of the control group were 4.94?0.81-5.36?0.97 in experiment two.Meantime the egg antigen levels in the injected mice might be lower.This study shows that the control of immunomodulation of granulomatous formation in the hosts injected with immunoenhance reagent "425" can be induced.