[Human cytotoxic T lymphocyte responses specific to the DNA vaccine of Wilms' tumor gene product].
OBJECTIVE
To 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.


METHODS
Synthesized 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.


RESULTS
The 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.


CONCLUSIONS
Using 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.
Zhang
Bin
B
Department of Internal Medicine, Hankou Hospital, Wuhan 430010, China. jincheng62@sohu.com
Wang
Zheng
Z
D000818:Animals; D016516:CD4-CD8 Ratio; D015496:CD4-Positive T-Lymphocytes / Q000166:cytology; D018414:CD8-Positive T-Lymphocytes / Q000166:cytology; D005260:Female; D016162:Genes, Wilms Tumor; D005822:Genetic Vectors; D006801:Humans; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D010957:Plasmids; D013602:T-Lymphocytes, Cytotoxic / Q000166:cytology / Q000276:immunology*; D019444:Vaccines, DNA / Q000235:genetics / Q000276:immunology*; D025721:WT1 Proteins / Q000235:genetics / Q000276:immunology; D023041:Xenograft Model Antitumor Assays
D019444:Vaccines, DNA; D025721:WT1 Proteins