基因转染增加免疫低下大鼠肺内γ-干扰素水平的研究  

STUDY OF INCREASING THE LEVEL OF IFN-γ IN THE LUNGS OF IMMUNOCOMPROMISED RATS WITH GENE TRANSFECTION

作　　者：马壮[1] 钱桂生[1] 黄桂君[1] 

机构地区：[1]第三军医大学新桥医院

出　　处：《解放军医学杂志》2002年第2期139-141,共3页Medical Journal of Chinese People's Liberation Army

摘　　要：探讨肺内基因转染、补充肺内IFN γ水平的可行性。构建重组大鼠IFN γ真核表达载体 ,转染免疫低下大鼠肺内。鉴定外源性质粒 ,并检测BALF和血清中IFN γ浓度和活性。结果发现 :构建的重组载体中IFN γ的基因序列与GeneBank中大鼠的IFN γcDNA序列相同。转染后BALF中细胞基因组DNA的PCR产物电泳可见转染的基因片段条带 ,转染后 2 1天仍可见其表达。pLXSN IFN载体组BALF中大鼠IFN γ浓度和活性显著高于pLXSN空载体组。血清中IFN γ浓度和活性二组间差异无显著性意义 (P >0 0 5 )。提示本研究构建的重组大鼠IFN γ真核表达载体可成功地转染大鼠肺内 ,整合入基因组DNA ,分泌有活性的IFN γ ,并较长期地表达。To study the possibility of enhancing IFN γ level in the lung of rats by gene transfection. The recombined IFN γ expression vector in eukaryotic cell was constructed, and it was transfected into lungs of immunocompromised rats. The expression of pLXSN IFN and pLXSN in the genomic DNA of the transfected cells was investigated, and the level and activity of IFN γ in the BALF and sera was assessed. The results showed that the sequence of the IFN γ gene in the constructed recombinant pLXSN IFN eukaryotic cell was the same as that of the IFN γ of rats found in the Gene Bank. The band of transfected plasmid was found when the eletrophoresis analysis of PCR product of genomic DNA of the transfected cells in BALF was done. It could still be the found 21 days after transfection. The level and activity of IFN γ in the BALF of the transfected pLXSN IFN group were markedly higher than those of transfected pLXSN group.On the other hand, the level and activity of IFN γ in sera showed no difference between the two groups. It was suggested that the constructed recombinant IFN γ exprossion vector in eukaryotic cell could be transfected into the cells of the lungs of rats successfully, inserted into the genomic DNA, resulting in active secretion of IFN γ, and prolongation of its expression.

关 键 词：干扰素Ⅱ型 免疫减低宿主 基因治疗 基因转染 呼吸道感染 大鼠 肺部感染 

分 类 号：R56[医药卫生—呼吸系统] R392.11[医药卫生—内科学]