摘要
本研究观察从人B细胞淋巴瘤细胞系基因组DNA及RNA分别构建的表达载体作为核酸疫苗诱导小鼠产生抗肿瘤免疫反应的情况。分别提取人B淋巴瘤细胞系Namalwa细胞的基因组DNA及RNA ,将免疫球蛋白重链可变区 (IgHV)基因片段克隆到pcDNA 3.0真核表达载体中作为独特型核酸疫苗 ,其中DNA来源的核酸疫苗用LipofectAMINE转染COS细胞 ,然后用RT PCR观察转录及剪接的情况。两种不同来源的核酸疫苗分别肌内注射免疫小鼠后 ,用间接免疫荧光法检测抗独特型抗体的生成。结果发现 ,基因组DNA来源的核酸疫苗在COS细胞中能成功地转录 ,转录产物大小为 4 77bp ,其中 86bp的内含子区域被剪接 ;两种疫苗免疫动物后均可诱导针对Namalwa细胞特异的抗独特型抗体 ,并于第 6周达高峰。结论提示 ,淋巴瘤基因组DNA和RNA来源的IgHV基因片段均可制备核酸疫苗 。
This study was to investigate the anti lymphocytic malignancy immunologic effects induced by two types of the idiotypic nucleic acid vaccines which were constructed from the genomic DNA and RNA of the human B lymphoma cell line respectively. Namalwa cell line and BALB/c mice were used as the models. The gene fragments of the IgH variable region(IgHV), which were obtained from the genomic DNA and RNA of Namalwa cell respectively, were cloned into the eukaryocytic expression vector pcDNA 3.0 to be used as the idiotypic nucleic acid vaccines. After transfecting COS cells with one of vaccines constructed from the genomic DNA by using LipofectAMINE,the result of transcription was identified by using RT PCR. The experimental mice were immunized by intramuscular injection with two types of vaccines. The specific anti idiotypic antibody was detected by indirect immunofluorescence assay. The results showed that the nucleic acid vaccine constructed from the genomic DNA can be transcribed in COS cells, the transcription product turned shorter,and the intron region of 86 bp was spliced accurately. When immunizing the mice, two vaccines both induced the anti idiotypic antibody against Namalwa cell, the anti idiotypic antibody could be detected since the fourth week after immunization, and got to the peak of titer on the sixth week. It was concluded that the nucleic acid vaccines against lymphoma can be constructed from both the genomic DNA and RNA.
出处
《中国实验血液学杂志》
CAS
CSCD
2002年第2期126-130,共5页
Journal of Experimental Hematology
基金
国家自然科学基金资助项目编号 39970 31 3
