Differential Patterns of Liver Proteins in Experimental Murine Hepatosplenic Schistosomiasis

Schistosoma mansoni eggs produced by adult worms in the mesenteric vasculature become trapped in the liver, where they induce granulomatous lesions and strong immune responses. Infected individuals suffer from intestinal schistosomiasis (INT) in 90% of cases, whereas the remaining 10% present with severe hepatosplenic schistosomiasis (HS). The CBA/J mouse model mimics human disease, with 20% of infected mice developing hypersplenomegaly syndrome (HSS) that resembles HS and 80% developing moderate splenomegaly syndrome (MSS) similar to INT. We studied differential patterns of protein expression in livers of 20-week-infected CBA/J mice with MSS or HSS to understand the molecular changes that underlie these two disease forms. Using differential in-gel electrophoresis to identify differentially expressed protein spots, we found 80 protein spots significantly changed with infection and 35 changes specific to severe disease. In particular, the abundances of prohibitin 2, transferrin isoforms, and major urinary protein isoforms were significantly altered in HSS mice. Furthermore, annexin 5, glutathione S-transferase pi class, and S. mansoni phosphoenolpyruvate carboxykinase expression levels changed significantly with schistosome infection. Additionally, levels of major urinary protein decreased and levels of transferrin increased significantly in the sera of HSS mice compared to levels in sera of MSS or control mice, and these differences correlated to the degree of splenomegaly. These findings indicate that the liver protein abundances differ between MSS and HSS mice and may be used for the development of diagnostic markers for the early detection of hepatosplenic schistosomiasis.Schistosomiasis affects 200 million people worldwide (13), and Schistosoma mansoni-associated hepatomegaly is estimated to affect 8.5 million people (62). From animal studies, it is estimated that the acute phase of infection for schistosomiasis occurs roughly 5 to 6 weeks after infection, when the parasitic eggs are swept into the liver microvasculature and induce granulomatous lesions. The chronic phase begins more than 12 weeks postinfection, with periportal liver fibrosis and extreme splenomegaly. In 90% of infected individuals, the egg-associated inflammation recedes, resulting in intestinal schistosomiasis (INT). In contrast, 10% of infected individuals present with severe hepatic and periportal fibrosis, portal hypertension, and portal-systemic venous shunts due to hepatic granulomas; these contribute to life-threatening hepatosplenic schistosomiasis (HS) (27). Granulomas play an important role in the development of the severe pathology of schistosomiasis (56). Previous studies have shown that the development of schistosomiasis causes liver dysfunction by altering metabolic processes through the upregulation or downregulation of enzymes (25, 67). Despite successful control measures for schistosomiasis in many countries, population growth and movement (13) and risk of reinfection (38) have played major roles in the transmission and spread of schistosomiasis to new areas.Our research focuses on an understanding of the protein patterns associated with the two distinct pathological forms of schistosomiasis. We used the CBA/J mouse model, as it reproduces human disease forms, with 20% of CBA/J mice developing hypersplenomegaly syndrome (HSS), which resembles HS, and 80% of the mice developing moderate splenomegaly syndrome (MSS), which is similar to INT (27). We have used two-dimensional differential in-gel electrophoresis (2D-DIGE) wherein protein samples are prelabeled with CyDyes before two-dimensional electrophoresis (2DE) for differential analysis, enabling the accurate tracking of qualitative and quantitative differences between MSS and HSS samples. Matrix-assisted laser desorption ionization (MALDI)-time of flight (TOF) mass spectrometry was used to identify protein spots that changed during the development of MSS and HSS. We believe that identifying disease-specific proteins may contribute to the early detection of and treatment strategies for hepatosplenic schistosomiasis.     

Tumour Necrosis Factor in Hepatosplenic Schistosomiasis

Periportal fibroplasia is the dominating feature of hepatosplenic schistosomiasis. Since monokines play an important role in the regulation of fibroplasia, tumour necrosis factor (TNF) and interleukin 1 beta (IL-1 beta) were assessed in sera and cell culture supernatants from patients with intestinal and hepatosplenic schistosomiasis before and 3-6 months after treatment with praziquantel. Uninfected controls were from the study area in Alagoas, Brazil. TNF was measured using an L-M mouse fibroblast bioassay and radioimmunoassays specific for TNF-alpha. Whereas TNF-alpha was elevated threefold in the patients' sera, three- to five-fold reductions of TNF were observed by radioimmunoassay and bioassay, respectively, in cell culture supernatants of hepatosplenic schistosomiasis patients. Significant deviations, in opposite directions, from TNF levels in control sera and supernatants are most plausible in the event of a sequestration of TNF-alpha-producing cells from the circulation. This process may be disease stage-specific since a dichotomy between incipient and advanced cases of hepatosplenic schistosomiasis became apparent in the amplitude and kinetics of changes during the follow-up after treatment.     

Hepatosplenic and other gammadelta T-cell lymphomas

The 2005 Society for Hematopathology/European Association for Haematopathology Workshop session 11 was dedicated to hepatosplenic T-cell lymphoma (HSTCL). HSTCL is a rare aggressive type of extranodal lymphoma characterized by hepatosplenomegaly, bone marrow involvement, and peripheral blood cytopenias. HSTCL exhibits a distinctive pattern of infiltration; tumor cells preferentially infiltrate the sinusoids of the splenic red pulp, liver, and bone marrow. The tumor cells have a nonactivated cytotoxic T-cell immunophenotype and frequently carry a recurrent cytogenetic abnormality, isochromosome 7q. Most cases express the gammadelta T-cell receptor, but cases can have an alphabeta phenotype and are considered to be a variant of the disease. Although HSTCL is the prototype peripheral T-cell lymphoma expressing the gammadelta T-cell receptor, non-HSTCL proliferations of gammadelta T cells can involve other extranodal sites, mainly skin and mucosa. These gammadelta T-cell lymphomas display marked heterogeneity in clinical and histologic features. In contrast with HSTCL, non-HSTCL gammadelta T-cell lymphomas frequently have an activated cytotoxic phenotype and most likely are not a single disease entity.     

肝脾γδT和αβT细胞淋巴瘤

从 1990年Ｆａｒｃｅｔ等描述了 2例肝脾γδＴ细胞淋巴瘤以来 ,人们渐渐认识到肝脾γδＴ细胞淋巴瘤是一个独立的淋巴瘤亚类 ,1994年它被ＲＥＡＬ分类确认为一个临时的淋巴瘤亚类[1] ;而在新的ＷＨＯ血液和淋巴组织肿瘤分类中它被确认为一个独立的疾病[1] 。它有典型的临床病理特征 ,包括多为青年男子发病 ,有Ｂ症状 (发热、夜汗、体重减轻 ) ,肝脾肿大 ,没有淋巴结病 ,重度贫血 ,明显的血小板减少症。患者多在诊断后 1年内死亡。组织学上 ,主要为小到中等大小的肿瘤细胞浸润脾脏的脾索和脾窦 ,以及肝窦 ,骨髓血窦等。特征性的免疫表型为Ｃ…     

黄芪注射液治疗糖尿病肾病的实验研究

目的:探讨黄芪注射液对糖尿病肾病(DN)的作用。方法:链尿佐菌素(STZ)腹腔注射诱导建立DN大鼠模型,检测体重、血糖、内皮素(ET)和一氧化氮(NO)的变化。结果:DN大鼠NO含量降低,体重、血糖、ET含量明显升高;黄芪注射液能减少DN大鼠体重、血糖和ET含量,提高NO水平。结论:黄芪注射液具有保护肾脏的作用。     

Emmunodiagnosis of Schistosomiasis

The most efficacious and practical means of diagnosing human schistosomiasis is based on the detection of infection-specific antibodies. Because of their high sensitivity and specificity, antibody assays remain the most practical assays for epidemiologic studies and patient management. Antibody assays are particularly useful in the diagnosis of schistosomiasis in visitors from developed countries to endemic areas. These patients are often lightly infected, and tests that depend on detection of ova or circulating antigens are not reliable for these type of light and acute infections. Initial screening may be performed in the field or laboratory with the FAST-ELISA, using adult microsomal antigens. Species-specific confirmation is obtained by immunoblots with the same antigens.     

Levamisole Restored the Compromized State of Immunity after Specific Chemotherapy in Experimental Schistosomiasis Mansoni

Mice infected for 45 days with 120 Schistosoma mansoni cercariae and treated with levamisole (25 mg/kg subcutaneously) have more efficient acquired immunity when challenged with 240 Schistosoma mansoni cercariae the same day of treatment (97.7% # 87.7% in infected challenged controls). In praziquantel-treated mice (500 mg/kg for 2 days orally), the reduction in the percent resistance (45.5%) was accompanied by a significant diminution in the size of granuloma, delayed foot pad swelling and granuloma proportionate T-helper cells number. Levamisole when given two weeks post praziquantel treatment and with the challenge infection increased the percent resistance to 79.2%. The increase in percent resistance recorded in mice receiving both praziquantel and levamisole was accompanied by restoration of granuloma size, delayed foot pad swelling and granuloma proportionate T-helper cells number to infected challenged untreated control values. Results reveal-beside efficacy of levamisole as immunoregulant in schistosome immunity - a possible role for the granuloma as a T-cell mediated response in maintenance of immunity.     

Schistosomiasis of the bladder

Two cases of the urinary bladder schistosomiasis are described. The role of the urinary bladder biopsy in the diagnosis of the disease is believed to be great.     

Hepatosplenic gammadelta T-cell lymphoma following seven malaria infections

Hepatosplenic gammadelta T-cell lymphoma (HSTL) is a clinicopathological entity associated with an immunocompromised status in approximately 25% of patients. Herein is described a case of HSTL in a 53-year-old Brazilian man with seven previous malaria infections, initially misdiagnosed as a hyperreactive splenomegaly due to chronic malaria. A characteristic lymphoid infiltrate was observed in spleen, liver and bone marrow sinusoids/sinuses. Neoplastic cells had a CD45RO+, CD2+, CD7+, CD3+, CD5-, CD8+, CD56+, perforin+, FasL-negative, T-cell receptor (TCR)alphabeta-negative, TCRgammadelta+ profile. Analyses of gamma and delta TCR rearrangements confirmed diagnosis of gammadelta T-cell lymphoma by detecting VgammaI/Vdelta1-Jdelta1 clonal rearrangements. Sensitive polymerase chain reaction (PCR) for Plasmodium falciparum, Epstein-Barr virus and herpesvirus-8 failed to demonstrate infection. The disease progressed to a fatal outcome following cutaneous infiltration and leukemic proliferation. The authors also comment on the association of lymphoma and infection, focusing on PCR diagnosis of TCRgamma and delta clonal rearrangements and the presumed pathogenic events leading to HSTL in the context of chronic malaria infection. Initial lymphomagenic stages might not be direct consequences of antigenic stimulation of Vdelta1 T-cells, but might depend on interactions between gammadelta T and B cells during cooperative or regulatory responses to Plasmodium sp.     

血吸虫病肝内微血管阻塞部位的研究

本研究采用血管墨汁灌注方法、血管塑料铸型方法,结合扫描电镜和透射电镜。观察了血吸虫病鼠(n=10)和兔(n=10)肝内微血管的变化。发现血吸虫病动物肝内门脉分支中断,肝窦数目减少,肝静脉分支扭曲、变形,肝窦毛细血管化,肝窦内皮细胞损伤,肝窦内有白细胞嵌塞。表明血吸虫病肝不仅有窦前梗阻,肝窦内及窦后亦有梗阻因素存在。