Prediction And Identification Of Epitopes In The Emy162 And TSP3 Antigen Of Echinococcus Multilocularis

Objective To identify and analyze B-cell and T-cell(Th1, Th2, Th17)epitopes of the antigen protein Emy162 and TSP3, Echinococcus multilocularis. To provide theoretical basis for the development of anti-Em multi-epitope vaccines. Methods 1. The secondary structure of the protein was analyzed using SOPMA server. The T-cell and B-cell epitopes were predicted using IEDB, Syfpeithi, Bcepred and ABCpred online software. 2. To collect the spleen lymphocytes of BALB/c mouse, which were immunized by Emy162 protein, TSP3 protein and PBS, and co-cultured the spleen lymphocytes stimulated with a chemically synthesized peptide antigen. The spleen lymphocyte proliferation assay and AE serum ELISA test were used to identify B-cell epitopes. 3. To stimulate mouse spleen lymphocytes with Synthetic T-cell peptides and detect the expression of Th1, Th2, Th17 cytokines by ELISA ELISpot, flow cytometry, to identify the Th1, Th2, Th17 epitopes. Results 1. Emy162, TSP3 proteins are potential advantages of antigenic epitopes. The high? scoring T? cell epitopes of Emy162 protein were located at positions E7-13、E36-41、E80-89、E87-96、E97-106、E129-139, whilst the likely B-cell epitopes were located at positions E7-13、E19-27、E28-36、E37-48、E78-83、E101-109、E112-121、E129-139. The high? scoring T? cell epitopes of TSP3 protein were located at positions T33-42、T45-55、T53-63、T68-77、T80-90、T92-104、T110-122、T134-144, whilst the likely B-cell epitopes were located at positions T18-33、T45-55、T53-63、T64-75、T80-90、T92-104、T110-122.2. Each B cell epitope of Emy162, TSP3 protein is different in antigenic.Three advanced antigen epitopes of Emy162 were E19-27 、 E112-121 、E129-139. Three advanced antigen epitopes of TSP3 were T18-33、T45-55、T110-122. 3. T-cell epitope of Emy162 protein can induce different expression of cytokines. Four Th1 advanced antigen epitopes of Emy162 were E7-13 、E36-41、E80-89、E129-139.Three Th2 advanced antigen epitopes of Emy162 were E36-41 、 E87-96 、 E97-106.Three Th17 advanced antigen epitopes of Emy162 were E36-41、E87-96、E97-106. 4. T-cell epitope of TSP3 protein can induce different expression of cytokines. Four Th1 advanced antigen epitopes of TSP3 were T33-42、T45-55、T80-90、T110-122.Three Th2 advanced antigen epitopes of TSP3 were T45-55、T68-77、T92-104.Two Th17 advanced antigen epitopes of TSP3 were T53-60 、 T80-90. Conclusions 1.There are advanced antigenicity and potential epitopes in Emy162 and TSP3 protein. Eight T cell and seven B cell dominant epitopes of the TSP3 antigen, six T cell and eight B cell dominant epitopes of the Emy162 antigen were revealed by the bioinformatics methods, which may be of use in the development of a dominant epitope vaccine. 2. There are respectively three dominant B-cell epitopes, four dominant Th1 epitopes, three dominant Th2 epitopes, three dominant Th17 epitopes, in Emy162 antigen. 3. There are respectively three dominant B-cell epitopes, four dominant Th1 epitopes, three dominant Th2 epitopes, two dominant Th17 epitopes, in TSP3 antigen. 4. The identification of dominant epitopes laid the theoretical foundation for subsequently research of efficient multi-epitope vaccine, used in immunization vaccination,diagnosis, treatment of AE.