论文部分内容阅读
用反转录ＰＣＲ的方法，从ＢＡＬＢ／ｃ小鼠脾细胞中扩增出Ｂ７－１ｃＤＮＡ后，插入ｐｃＤＮＡ３质粒中构建成小鼠Ｂ７－１ｃＤＮＡ的真核表达载体ｐＣＤ－ｍＢ７．１，经酶切鉴定和序列分析证实此表达载体中插入的Ｂ７－１ｃＤＮＡ的序列与文献报道一致．通过脂质体介导将ｐＣＤ－ｍＢ７．１导入Ｂ７－１的小鼠黑色素瘤细胞系Ｂ１６（Ｆ０）中，经ＲＴ－ＰＣＲ和ＲＮＡ斑点杂交初步证实Ｂ７－１在肿瘤细胞中获得了稳定有效的表达．同源小鼠脾淋巴细胞与肿瘤细胞混合培养后采用ＬＤＨ释放改良法测定淋巴细胞特异杀伤活性，结果显示，与野生型和模拟转染的Ｂ１６细胞相比，Ｂ７－１基因转染的Ｂ１６细胞能较有效诱导淋巴细胞产生针对野生型Ｂ１６细胞的特异杀伤活性（ｐ＜０．０２）．这说明，将Ｂ７－１基因导入肿瘤细胞表达能提高其免疫原性，诱导有效的抗肿瘤免疫反应
B7-1 cDNA was amplified from BALB / c mouse spleen cells by reverse transcription PCR and inserted into pcDNA3 plasmid to construct the eukaryotic expression vector pCD-mB7.1 of mouse B7-1 cDNA. Identification and sequence analysis confirmed that the expression vector inserted B7-1 cDNA sequence consistent with the literature. PCD-mB7.1 was introduced into B7-1 mouse B16 cell line (F0) by lipofectamine. The results of RT-PCR and RNA dot blot showed that B7-1 was stable and effective in tumor cells expression. Lymphocyte-specific cytotoxic activity of splenic lymphocytes from mice homogenized with tumor cells mixed with tumor cells was measured by LDH release modification. The results showed that compared with wild-type and mock-transfected B16 cells, B7-1-transfected B16 cells The specific killing activity of lymphocytes against wild-type B16 cells was more effectively induced (p <0.02). This shows that the B7-1 gene into tumor cell expression can improve its immunogenicity, induce effective anti-tumor immune response