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Virologica Sinica, 19 (5) : 439-443, 2004
Research Article
Significance of BLCL Constructed from PBMC in HFRS Patients
Tangdu Hospital, The Fourth Military Medical University, Xi’an 710038,China
In order to provide some data for the study on T cell epitopes of Hantaan virus nucleocapsid protein, we constructed EBV-transformed B lymphoblastoid cell line (BLCL) from HFRS patients’ PBMC. We inserted HTNV S and 5’ and 3’terminals of S gene segment into a eukaryotic expression vector pcDNA 3.1/V5-His TOPO. The recombinant expression plasmids pcDNA3.1-S and pcDNA 3.1-S-N pcDNA3.1-S-C were constructed. BLCL were transfected in vitro with them, selected by G418 and used as target cells for CTL cytotoxicity assay. The expression of HTNV S and 5’ and 3’terminals of S gene segment in BLCL was detected by indirect immunofluorescence assay (IFA). These recombinant plasmids can be expressed efficiently and steadily in BLCL. We can provide a tool for the study on T cell eptitopes of Hantaan virus nucleocapsid protein. The CTL clone recognized and killed the target cells with specificities of HTNV-NP. Cytotoxicities(%) were 50.2%, 39.0% and 25.4%, respectively. HTNV T cell antigenic epitopes were mainly localized on the C-terminal of the virus nucleocapsid protein.
  Published online: 20 Oct 2004
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