| Objective: Epstein-Barr virus is associated with a variety of benign and malignant diseases in humans,such as mononucleosis,Burkitt’s lymphoma,Hodgkin’s lymphoma,nasopharyngeal carcinoma,and post-transplant lymphoproliferative disorders.However,research on EBV and related diseases has been slow due to the lack of suitable animal models.Tree shrews are small emerging experimental animals closely related to primates and have great potential for studying human diseases.Our team has initially established an animal model of EB virus-infected tree shrews in the previous research.Based on the previous research,this study further explored the EB virus model of tree shrew infection.We carried out immunosuppression by inoculating Epstein-Barr virus on tree shrews,and detected the expression and histopathological changes of EB gene and protein,and explored whether the Epstein-Barr virus has similar biological characteristics to humans in tree shrews.Whether tree shrews can be used as an animal model of EB virus-related diseases.Methods: Twenty healthy adult tree shrews were selected,16 in the experimental group and 4 in the control group.In the experimental group,1 mlof 1.2×108 copies/ml EB virus solution was administered through the nose and mouth inoculum.The control group was inoculated with 1 ml of normal saline in the same manner.The experimental group and the control group were treated with cyclosporine A 20 mg/kg/day for 6 weeks.Injection for 6 weeks.Tree shrews regularly collect blood from the femoral vein,separate peripheral blood mononuclear cells and serum;dissect tree shrews and collect important organ tissues.We performed peripheral blood leukocyte counts on tree shrews;real-time quantitative PCR was used to detect Epstein-Barr virus copy number in PBMCs;RT-PCR was used to detect the expression of Epstein-Barr virus-related genes in PBMCs;ELISA was used to detect changes in anti-EBV-related antibodies in serum.The histopathological changes were detected by HE staining.The expression of Epstein-related proteins in tissues was detected by immunohistochemistry.The expression of EBERs in tissues was detected by in situ hybridization.The expression of EBNA1 protein in spleen was detected by Western blot.Result:Peripheral blood white blood cell count: Tree shrew showed a significant downward trend after immunosuppression,and slowly rose after stopping immunosuppression;Real-time quantitative PCR results:After the immunosuppression of the experimental group,all the animals in the experimental group had an obvious increase in the number of Epstein-Barr virus copies.When the immunosuppression was stopped,the copy number showed a downward trend;RT-PCR results: The expression of EBNA1,EBNA2,EA and BZLF1 in the experimental group increased after immunosuppression;HE staining results: Some experimental groups showed EB virus-associated hyperplasia in tissues such as spleen and lymph nodes,and some tree shrews showed EB virus-associated lymphoproliferative diseases;The results ofimmunohistochemistry: LMP1 expression was detected in the spleen,lymph nodes,liver,lung,bone marrow and other tissues of the experimental group;Results of EBERs: EBERs were detected in the spleen,lymph nodes,liver,lung,bone marrow and other lesions of the experimental group.Western-blot results: In the experimental group,except for Ts16,the expression of EBNA1 was detected in other spleen tissues.Conclusion:Tree shrews can be infected with Epstein-Barr virus by oral and nasal inoculation.After injection of CsA for immunosuppression,Epstein-Barr virus can be reactivated.After immunosuppression,tree shrews produce a similar EB virus-associated lymphoproliferative disease with potential as an animal model of EB virus infection and related diseases. |
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