M-CSF和IFN-γ基因转染的巨噬细胞的体外生物学特性

THE IN VITRO IMMUNOLOGICAL CHARACTERISTICS OF MCSF AND IFNγ GENESTRANSFECTED MACROPHAGES

利用缺陷型腺病毒的介导将与巨噬细胞功能密切相关的细胞因子Ｍ－ＣＳＦ和ＩＦＮ－γ基因联合转入小鼠腹腔巨噬细胞，对其体外生物学特性进行了观察。结果表明：重组腺病毒能高效转染小鼠腹腔巨噬细胞，其转染效率高达９０％，ＲＴ－ΡＣＲ结果显示Ｍ－ＣＳＦ和ＩＦＮ－γ基因已成功地转入巨噬细胞，其培养上清中有高水平的目的因子分泌，基因转染后的巨噬细胞扫描电镜下细胞表面泡状突起、枝状突起增加，基因转染的巨噬细胞处于Ｇ１期的细胞比例显著减少，Ｓ期的细胞比例显著增加。该结果提示：Ｍ－ＣＳＦ和ＩＦＮ－γ有助于巨噬细胞的体外扩增和功能维持。

MCSF and IFNγ are two cytokines which can regulate the functions of Mφs.In the present study,mice peritoneal Mφs were transfected with MCSF and IFNγ genes by recombinant adenovirus and their biological characteristics were observed.Xgal staining showed that the freshly isolated Mφs could be transfected effectively (up to 90%)by a replication defective adenovirus harboring LacZ gene.The results of RTPCR showed that MCSF and IFNγ genes were successfully transfected into Mφs.High levels of MCSF or IFNγ could be detected in the supernatants of Mφs 4h after gene transfection and the secretion persisted for more than 10 days.Under scanning electron microscope(SEM),surface of normal and LacZ gene transfected Mφs were smooth while that of Mφ transfected with MCSF and IFNγ genes showed surface ridges or ruffles and fine fingerlike processes or filopo dia.Cell cycle analysis revealed that the percentage of Mφs in G1 stage decreased while that in S stage increased after MCSF and/or IFNγ genes transfection.The increasement of the cell proportion in S stage showed that Mφ cotransfected with MCSF and IFNγ genes were in the form of proliferation.The results indicated that MCSF and IFNγ genes transfection would expand the Mφ in vitro and maintain their functions which would contribute to the antitumor effects of MCSF and IFNγ gene transfected Mφ.