摘要
目的:获得表达HLA-DR基因的小鼠黑色素瘤细胞。方法:应用RT-PCR技术从人B淋巴瘤细胞系Raji中克隆了HLA-DR3α和β链cDNA,并经测序证实。将其分别插入至真核表达载体pLXSN和pCEP4中,用脂质体转染小鼠黑色素瘤细胞B16,然后用G418和Hyg双重筛选。结果和结论:经G418和Hyg双重筛选后获得的抗性细胞经流式细胞仪检测,表明59%的细胞表达所转染的HLA-DR3分子。PCR从其基因组DNA中扩增到α和β链基因,提示HLA-DR3cDNA已整合至宿主细胞的基因组DNA中。本研究为研究异种MHCⅡ转染在肿瘤免疫中所起的作用奠定了基础。
Objective: Obtaining mouse melanoma cells expressing HLA DR genes. Methods: HLA DR3 α And β chain cDNA were cloned by RT PCR from human B lymphoma Raji cell line and confirmed by DNA sequencing. The recombinant expressing vectors were constructed by inserting the α and β chain cDNA into pCEP4/pLXSN vectors respectively. Mouse melanoma B16 was transfected with the recombinant vectors by lipofectamine and selected with G418 and Hyg. Results and Conclusion: After G418 and Hyg selecting, the HLA DR3 transfected clones were obtained. Approximately 59% of the transfected cells expressed HLA DR when assayed by FACS. It was confirmed that the HLA DR3 cDNA had been integrated into the genomic DNA of B16 DR cells by PCR analysis. Our study enables us to further investigate the role of xenogeneic MHCⅡ in tumor gene therapy.
出处
《军事医学科学院院刊》
CSCD
北大核心
1998年第4期253-256,共4页
Bulletin of the Academy of Military Medical Sciences