Construction of TCR V_? Nucleic Acid Vaccine and Testing on Its Biological Effects / 中国肿瘤生物治疗杂志

ABSTRACT

Objective: To construct TCR V? nucleic acid vaccine. Methods: A fragment of TCR V? 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? gene from Jurkat cells was obtained and proved to be identical to published sequence. A recombinant plasmid pcDNA3-TCR V? was comstructed. Its expression was detectable on mR- NA level after being transfected into SP2/0 cells. Antiserum from mice immunized with pcDNA3-TCR V? reacted strongly with Ju- rkat cells and SP2/0 cells transfected by pcDNA3-TCR V?, while shows no reaction on CEM cells expressing TCR?? and SP2/0 cells. Antisera from normal force and mice immunized with pcDNA3 were both negative on Jurkat cells. The results of immunocy- tochemistry indicated that BALB/c mice immunized with pcDNA3-TCR V? produced specific antibody to Jurkat TCR V?.Conclu sion The TCR V? nucleic acid vaccine we constructed can induce humoral immunity.