| Schistosomiasis is a serious threat to the public health worldwide. Egg granuloma formation and subsequently hepatic fibrosis are the main pathological basis of schistosomiasis Japonica. When it's impossible to restrain schistosoma infection completely,a new way of controlling schistosomiasis is anti-pathology immunity,by mean of inoculating anti-pathology vaccine of schistosomiasis,could lessen the pathological damage of host as much as possible and improve the condition of organism. Guan Xiaohong,et al. (1991) established a strain of monoclonal anti-idiotypic antibody NP30 of Schistosoma japonicum,whice is an internal image antigen of gut associated antigen (GAA),its antibody isotype is IgM. NP30 has a partial cross-reaction with soluble egg antigen (SBA) and membrane associated antigen (MAA). The mouse model of hepatic egg granuloma of Schistosoma japonicum confirmed that NP30 has sensitizing effect on formation of hepatic egg granuloma,active immunity in sheeps and mice with NP30 can induce good protection against infection with Schistosoma cercariae and reduces immunopathologic damage to host,NP30 plays a significant down-modulatory role to hepatic granuloma and fibrosis. NP30 is regard as a candidate molecules of anti-pathology vaccine against schistosomiasis,however,anti-pathology immunity mechanism ofNP30 is still unclear. Therefor,in this article,we studied the effect of NP30 on apoptosis of egg granuloma cells in mice infected with Schistosoma japonicum, and explore the molecular regulating mechanism of NP30. Methodsl.Both sexes of 6-8 week old (weight 18-22 g) BALB/c mice were randomly divided into three groups,Experimental group:'Hie experimental group had twenty mice. Mice were immunized three times,separated by 2-week intervals,by intraperitoneal injection of NP30 (10 |ii g/dose/mouse). Four week after final boosting,vaccinated and control mice were challenged percutaneously with 12 S. japonicimi cercariae (Chinese mainland strain) by the cover glass method. Control group:The control group had twenty mice and was injected normal saline intraperitoneally. The times and intervals of immunization were the same as the experimental group. Mice of the experimental group and control group were respectively killed at the 39th,49lh 64th,108th,112th day after challenged with cercariae. Normal control group:Normal control group had four mice and was without any treatment.2. The morphology of apoptotic cells in hepatic egg granuloma of both experimental group and control group were studied by transmission electron microscopy.3. We employed TUNEL method to analyze apoptotic egg granuloma cells in experimental group and control group by incorporating digoxigenin-conjugated dUTP into the DNA fragment with the TdT.4. Apoptotic rate of egg granulomous cells in three groups wasdetected by Flow Cytometry (FCM),the histogramic analysis of DNA contents revealed the appearance of hypodiploid DNA peak immerged befors G! phase,regarded as Apo peak.5. The expressions of apoptosis-related gene bax,bcl-2,c-fos,death receptor fas and fasL(fas Ligand) were examined by immunohistochemistry (UltraSensitiveTM S-P). bax and fas gene were investigated by the DIG-labelled oligodeoxynucleotide probe of in-situ hybridization.6. By Computer Image Analysis System NYD-1000 analysing the results of TUNEL and immunohistochemistry,accounting the size and positive cell of egg granulomas,apoptotic and positive index of granuloma cell was determined .Results1. At the 39th and 49th day after infection ,the typical apoptotic cells of egg granulomas could be observed in control group. The morphologic feature of apoptotic cell included shrinkage,karyopyknosis and condensation of chromatin along with the membrane,progressive formation of apoptotic bodies. In experimental group,Many apoptotic cells could observed in egg granulomas of each time point after infection.2. The result examined by TUNEL comfirmed that egg granuloma cells occurred apoptosis. At...
|
PDF Full Text Request