Identification Of T Cell Epitopes Specific For Nucleocapsid Protein Of Hantaan Virus

Objectives: to identify the CD4~+T cell and CD8~+T cell epitopes specific for nucleocapsid protein of Hantaan virus.Methods: (1) PBMCs from HFRS patients were stimulated with 70 overlapping 15-mer peptides, and the positive response to peptides or peptide pools stimulation was tested using ELISPOT. (2) EBV-transformed B lymphoblast cell lines (B-LCL) were generated from PBMC of HFRS patients. CD4~+ T cells or CD8~+ T cells isolated from PBMC using Dynabeads were incubated with 15-mer peptide-loaded autologous 3-LCL overnight to determine if the stimulated CD4~+ T cells or CD8~+ T cells can secrete IFN-γ. The peptides which can elicit CD4~+ T cell responses were CD4~+ T cell epitopes, while the peptides which can elicit CD8~+ T cell responses contained one or more CD8~+ T cell epitopes. (3) The CTL epitopes encompassed within the 15-mer peptides which can elicit CTL responses in HFRS patients were predicted using three algorithms BIMAS, SYFPEITHI and ANN model. The...