Construction of TCR V β Nucleic Acid Vaccine and Testing on Its Biological Effects
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Abstract:
To construct TCR V β nucleic acid vaccine. Methods: A fragment of TCR V β gene was amplified by RT-PCR from a human T lymphoma cell line, Jurkat, and cloned into eukaryotic expression plasmid pcDNA3 by gene recombination. After sequencing, the recombinant plasmid was transfected into SP2/0 cells and its expression was detected on mRNA level. BALB/c mice were immunized with pcDNA3 and the recombinant plasmid by intramuscular routes. Antiserum was collected and detected by immunocytochemistry method. Results: A fragment of TCR V β gene from Jurkat cells was obtained and proved to be identical to published sequence. A recombinant plasmid pcDNA3-TCR V β was constructed. Its expression was detectable on mRNA level after being transfected into SP2/0 cells. Antiserum from mice immunized with pcDNA3-TCR V β reacted strongly with Jurkat cells and SP2/0 cells transfected by pcDNA3-TCR V β, while shows no reaction on CEM cells expressing TCRγδ and SP2/0 cells. Antisera from normal mice and mice immunized with pcDNA3 were both negative on Jurkat cells. The results of immunocytochemistry indicated that BALB/c mice immunized with pcDNA3-TCR V β produced specific antibody to Jurkat TCR V β. Conclusion: The TCR V β nucleic acid vaccine we constructed can induce humoral immunity.