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目的：制备Ｔ淋巴瘤ＴＣＲＶβ核酸疫苗。方法：利用ＲＴ－ＰＣＲ方法扩增出人Ｔ淋巴瘤细胞Ｊｕｒｋａｔ的ＴＣＲ Ｖβ基因片段，并将其重组入真核表达载体ｐｃＤＮＡ３。用重组表达载体转染ＳＰ２／０细胞，在ｍＲＮＡ水平上检测其表达。再用ｐｃＤＮＡ３和重组载体分别免疫ＢＡＬＢ／ｃ小鼠，收集抗血清，用免疫组化技术检测抗体产生情况。结果：扩增出ＴＣＲ Ｖβ的基因片段，测序结果证实其序列与已发表的一致。并构建出重组表达载体ｐｃＤＮＡ３－ＴＣＲ Ｖβ。该质粒转染到 ＳＰ２／０细胞，可在ｍＲＮＡ水平上检测到其表达。在用ｐｃＤＮＡ３－ＴＣＲＶβ免疫的小鼠抗血清中检测到抗Ｊｕｒｋａｔ细胞 ＴＣＲ Ｖβ的抗体。免疫组化结果表明，该血清不与表达 ＴＣＲγδ的人Ｔ淋巴瘤细胞发生反应，而与转染该基因的ＳＰ２／０细胞产生强阳性反应。正常小鼠血清与ｐｃＤＮＡ３免疫的小鼠血清与Ｊｕｒｋａｔ细胞不产生显色反应。结论：所制备的Ｔ淋巴瘤ＴＣＶβ核酸疫苗能有效地诱导机体产生体液兔疫反应。
Objective: To prepare TCRVβ DNA vaccine for T lymphoma. Methods: The TCR Vβ gene fragment of human T lymphoma cells Jurkat was amplified by RT-PCR and recombined into eukaryotic expression vector pcDNA3. SP2 / 0 cells were transfected with the recombinant expression vector and its expression was detected at mRNA level. Then, BALB / c mice were immunized with pcDNA3 and recombinant vector respectively to collect antiserum. Antibody production was detected by immunohistochemistry. Results: The gene fragment of TCR Vβ was amplified and confirmed by sequencing. The sequence was consistent with the published one. And construct a recombinant expression vector pcDNA3-TCR Vβ. The plasmid was transfected into SP2 / 0 cells and its expression was detected at the mRNA level. Antibodies to TCR Vβ against Jurkat cells were detected in mouse antisera immunized with pcDNA3-TCRVβ. The results of immunohistochemistry showed that the serum did not react with human T lymphoma cells expressing TCRγδ, but strongly positive with SP2 / 0 cells transfected with this gene. The mouse serum immunized with normal mouse serum and pcDNA3 did not develop a color reaction with Jurkat cells. Conclusion: The prepared T lymphoma TCVβ nucleic acid vaccine can effectively induce the body to produce the humoral immune response.