Expressing Location Of EWS-FLI1 In Ewing's Sarcoma Cells And Epitopes Prediction Of Fusion Region Of EWS-FLI1

ObjectiveInvestigate the expressing location of EWS-FLI1 fusion protein in Ewing's sarcoma cells , predict the T cell epitopes and B-cell epitopes of fusion region of EWS-FLI1, which provide experimental indiction for the next research.MethodsPerform immunocytochemical experiment with specific monoclonal antibody FLI1 after subculturing Ewing's sarcoma cell lines A673, analyze the expressing location of EWS-FLI1 in EWS cells.Apply bioinformatic softwares,such as BIMAS, SYFPEITHI, Predep, Immune Epitope Database and analysis Resource (IEDB) to predict most likely HLA-A * 0201-restricted CTL epitopes of fusion region.Analyze the secondary structure, the hydrophilicity, the accessibility, the antigen index of EWS-FLI1 fusion region,predict most likely B epitopes of fusion region with SOPM / SOPMA, Chou-Fasman ,Karplus-Schulz, Parker, Emini and Kolaskar-Tongaonkar softwares.ResultsImmunocytochemical staining proves that the expressing of EWS-FLI1 locating in the nucleus of EWS cells is about 84.78 percent in all the observed EWS cells, the expressing of EWS-FLI1 locating in cytoplasm of EWS cells is about 11.71 percent, the expressing of EWS-FLI1 locating in the membrane of EWS cells is about 3. 51 percent .Comprehensive analysis indicates that there are 9 most probable HLA-A * 0201-restricted CTL epitopes of fusion region after prediction by four kinds of T cell epitope predictive software.Analyze the secondary structure, the hydrophilicity, the accessibility, the antigen index of EWS-FLI1 fusion region,comprehensive analysis inducts that there exist most probable B epitope of fusion region after prediction by B cell epitope predictive softwareConclusionsEWS-FLI1 fusion protein mainly locates in the nucleus of EWS cells, and fewly locates in the cytoplasm and membrane of EWS cells.Apply predictive softwares can help to predict HLA-A * 0201-restricted CTL epitopes and B cell epitopes of fusion region successfully, which help for further research of EWS-FLI1 characteristics and for developing compound epitopes vaccine of EWS.