OBJECTIVETo construct the eukaryotic expression vector harboring Wilms' tumor gene (WT1y) and assess the cytotoxic T lymphocyte (CTL) response specific to the DNA vaccine of WT1y product in Balb/c mice and in vitro cultured cells.

METHODSSynthesized WT1y gene (321 bp including HLA-A*2402 anchoring residue) was cloned into the eukaryotic vector to construct the plasmid pCDNA3.1(+)/WT1y. WT1-specific antibody was detected in mouse serum by ELISA, and T lymphocyte proliferation was detected by flow cytometry. The CTL response was detected by co-culture of the murine spleen cells with the tumor cells.

RESULTSThe recombinant plasmid was confirmed using DNA sequencing. WT1-specific antibodies were detected in the serum of immunized mice, which showed significantly greater T lymphocyte proliferation than the control group (Plt;0.01). The CTLs resulted in specific lysis of WT1-expressing murine tumor cells.

CONCLUSIONUsing HBVC as the vector for WT1y gene, wenave obtained the virus-like particles that induce high cellular immune response, which shed light on a new approach for treatment of Wilms' tumor.