Establishment of Hantaan virus-specific T cell clone and its target cells in patients with hemorrhagic fever with renal syndrome
ObjectiveTo establish T cell clone which is specific for nucleocapsid protein (NP) of Hantaan virus (HTNV) in patients with hemorrhagic fever with renal syndrome (HFRS) and to lay a foundation for the study of T cell epitope and T cell immunology. MethodsPeripheral blood mononuclear cells (PBMCs) of patients with HFRS separated by Ficoll-Hypaque density gradient centrifugation were stimulated in vitro with inactivated HTNV and recombinant human IL-2. T cell clones were established with limiting dilution technique and clone phenotype was identified by flow cytometer. PBMCs infected by EB virus (EBV) were used to establish immortalized B lymphoblastoid cell lines (B-LCLs). B-LCLs infected by rVV were used as the target cells of HTNV-specific CTL clones. The antigenic specificity of T cell lines was identified by cytotoxicity assays. ResultsB-LCLs expressed NP were recognized by HTNV-specific T cell clones. Of the 5 individuals tested,three had high specific lysis rate. Five T cell clones were obtained from two patients and CD8 + T cell clone takes more than 60%. ConclusionHTNV NP- specific T cell clones and their target cells have been established. NP is potentially a major target of the CTL response during HTNV infection.