Study On The In Vitro Model Of Peripheral Blood Mononuclear Cells Of Ebv Infected Tree Shrews

Epstein-Barr virus(EBV)is currently known to be closely related to a variety of malignant tumors such as Hodgkin’s disease,nasopharyngeal carcinoma(NPC)and Burkitt’s lymphoma,but there is still a lack of related typical and ideal Experimental animal models,therefore,for the study of the characteristics of EBV,especially EBV in the process of tumorigenesis and development still need to be further explored,these are of great significance for the development of diseases and the development and application of drug treatment.At present,there have been studies showing that EBV can infect normal human B lymphocytes in vitro and can transform them into immortalized lymphoblastoid cell lines(LCLs).This study is based on this characteristic of EBV,by studying EBV in vitro Peripheral blood mononuclear cells(PBMCs)infected with tree shrews,rabbits,and rats were compared to investigate the potential of an EBV-infected tree shrew model in order to further study the characteristics and pathogenesis of EBV infection Lay the foundation.OBJECTIVE:In this study,EBV was used to infect the peripheral blood mononuclear cells of tree shrews in vitro to study their infection status and characteristics,and explore the feasibility of tree shrews as an EBV infection model in vitro research,which laid the foundation for further research on the characteristics and pathogenic mechanism of EBV infection.METHODS:Twenty adult tree shrews were selected and 2.5 ml of blood was collected from the femoral vein in awake state;peripheral blood mononuclear cell separation fluid was used to separate the peripheral blood mononuclear cells of tree shrews.Adjust the cell density to about 1×10~6/ml culture,add EBV virus suspension to the experimental group,and adjust the final concentration to 1×10~9 copies/ml;add the same amount of RPMI1640culture medium to the blank group.After adding EBV,regularly use a microscope to observe changes in PBMCs such as cell morphology,complete cell count,real-time fluorescence quantitative PCR technology to detect changes in EBV copy number in cells,RT-PCR to detect the expression of EBV-related genes,and immunofluorescence to detect the expression of EBV-related proteins In the case of transmission electron microscopy to observe whether the cells contain virus particles.RESULTS:After EBV infection,most tree shrew PBMCs can be successfully infected by EBV.The tree shrew PBMCs successfully infected by EBV showed agglomeration,morphology became larger and rounded,etc.;the number of tree shrew PBMCs cells after EBV infection increased,and the cells began to proliferate again,the growth rate reached the highest value within 2hours;EBV in cells after infection The viral load copy number rises,fluctuates,and finally maintains a certain concentration;the infected tree shrew PBMCs can detect the expression of EBV-related genes and related proteins.Transmission electron microscopy can observe the presence of herpes-like virus particles in infected cells.Conclusion:Epstein-Barr virus can infect the peripheral blood mononuclear cells of tree shrews in vitro.After infection,the cells have the potential to become EBV-related in vitro models;the survival time of the infected cells in vitro is prolonged and has some characteristics of LCLs.