摘要
目的 探讨以Ki ras基因 12位密码子点突变为靶点 ,构建DC Ad Ki ras(V12 )肺癌疫苗及其抗肺癌免疫治疗的可能性。方法 应用重组腺病毒的方法将人Ki ras基因 12位密码子点突变 (由甘氨酸转变为缬氨酸 )的cDNA导入DC ,构建DC Ad Ki ras(V 12 )肺癌疫苗 ,以流式细胞术、PCR、MLR和CTL等方法检测其免疫活性。结果 ( 1)DC Ad Ki ras(V12 )肺癌疫苗不仅能表达人Ki ras(V12 )基因 ,且能明显刺激T细胞增殖及提高CTL的杀伤作用。 ( 2 )突变Ki ras基因修饰的DC免疫小鼠后 ,可产生针对表达Ki ras(V 12 )基因的Lewis肺癌的特异性杀伤作用 ,但对B16黑色素瘤细胞则无明显杀伤作用。结论 以人突变Ki ras基因修饰的肺癌DC疫苗可在体外特异性诱导抗可表达Ki ras(V 12 )基因的Lewis肺癌的活性。
Objective To construct a DC-Ad-Ki-ras(V12) vaccine and investigate the anti-tumor activities of lung cancer dendritic cell vaccine modified by mutant Ki-ras gene in vitro. Methods Ki-ras(V12) cDNA was transfected into cultured bone marrow-derived DC with the recombinant adenovirus [(Ad-Ki-ras(V12)] containing human mutant Ki-ras gene. Anti-tumor activity of the vaccine was studied in vitro by flow cytometry, PCR, MLR and cytotoxicity assay. Results (1) The DC vaccine was confirmed not only to express Ki-ras(V12) gene, but also to remarkably stimulate lymphocyte proliferation and improve CTL activity. (2) The DC vaccine modified by mutant Ki-ras gene could induce specifical CTL activity of immunized mice against Lewis lung carcinoma that could express Ki-ras(V12) gene, but not to B16. Conclusion The DC vaccine modified by mutant Ki-ras gene can induce obvious anti-tumor activities against Lewis lung carcinoma that can express Ki-ras(V12) gene.
出处
《中国肺癌杂志》
CAS
2004年第2期104-107,共4页
Chinese Journal of Lung Cancer
基金
国家自然科学基金 (No .30 0 0 0 2 0 6)
高等学校博士点基金(No .2 0 0 0 55)资助~~