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目的通过复制缺陷型腺病毒载体的介导,将金黄色葡萄球菌肠毒素A(SEA)基因转染入C57BL/6小鼠淋巴细胞中,观察基因表达产物对B16细胞的作用。方法通过MTT法直接测定重组腺病毒颗粒感染的淋巴细胞的增殖情况,采用双抗体夹心ABC-ELISA法,测定淋巴细胞培养上清液中白介素2的水平。MTT法测定活细胞数目,以了解活化的C57BL/6小鼠淋巴细胞对B16细胞的杀伤效应。结果加入不同滴度的重组腺病毒颗粒后,C57BL/6小鼠淋巴细胞明显增殖;同时淋巴细胞培养上清液中白介素2水平也明显增加。重组腺病毒颗粒转染的C57BL/6小鼠淋巴细胞可明显杀伤B16细胞,效/靶比越高,杀伤效应越明显。结论SEA基因可以在C57BL/6小鼠淋巴细胞中表达,并且表达产物能够活化C57BL/6小鼠淋巴细胞,进而杀伤B16细胞。
OBJECTIVE: To investigate the effect of gene expression products on B16 cells by transfection of staphylococcal enterotoxin A (SEA) gene into lymphocytes of C57BL / 6 mice mediated by replication-defective adenovirus vector. Methods The proliferation of lymphocytes infected by recombinant adenovirus particles was directly determined by MTT method. The interleukin 2 level in lymphocyte culture supernatant was determined by double antibody sandwich ABC-ELISA. The number of viable cells was determined by MTT assay to understand the killing effect of activated C57BL / 6 mouse lymphocytes on B16 cells. Results After adding different titer of recombinant adenovirus particles, the lymphocytes of C57BL / 6 mice proliferated significantly. At the same time, interleukin 2 level in lymphocyte culture supernatants also increased significantly. Recombinant adenovirus particles transfected C57BL / 6 mouse lymphocytes can significantly kill B16 cells, the higher effect / target ratio, the more obvious the killing effect. Conclusion The SEA gene can be expressed in C57BL / 6 mouse lymphocytes and the expression product can activate C57BL / 6 mouse lymphocytes and then kill B16 cells.