Radzioch D, Hudson T, Boulé M, Barrera L, Urbance J W, Varesio L, Skamene E
Department of Medicine, Montreal General Hospital Research Institute, Quebec, Canada.
J Leukoc Biol. 1991 Sep;50(3):263-72. doi: 10.1002/jlb.50.3.263.
Congenic strains of mice susceptible (B10A.Bcgs) or resistant (B10A.Bcgr) to BCG were established. Here we describe the model system which has been established to analyze the functional activities of macrophages in the two strains. We have immortalized bone marrow macrophages from B10A.Bcgs and B10A.Bcgr congenic strains of mice and derived cloned macrophage lines designated B10S and B10R, respectively. B10R and B10S cell lines exhibited surface markers and morphology typical of macrophages. B10S and B10R were similar in their phagocytic activity, in their level of c-fms, in their transforming growth factor beta (TGF beta) mRNAs expression, and in their expression of tumoricidal activity in response to interferon-gamma (IFN gamma) plus lipopolysaccharides (LPS). However, B10R macrophages expressed a higher level of la mRNA when activated with IFN gamma compared with B10S macrophages. Analysis of the bacteriostatic activity of the two cell lines revealed that B10R macrophages were much more active in inhibiting Mycobacterium smegmatis replication than B10S. To measure the intracellular destruction of bacilli, a bactericidal assay based on hybridization with an oligonucleotide probe specific for mycobacterial ribosomal RNA was designed. The results demonstrated that B10R macrophages were endowed with enhanced constitutive bactericidal activity as compared with B10S. In conclusion we have obtained macrophage lines from bone marrow of B10A.Bcgs and B10A.Bcgr mice that express to a similar extent functional and phenotypic characteristics of macrophages. However, we demonstrate that relative to B10S macrophages, the B10R macrophages have higher expression of la mRNA and that they are constitutively more active in expressing mycobactericidal activity.
建立了对卡介苗敏感（B10A.Bcgs）或耐药（B10A.Bcgr）的近交系小鼠。在此，我们描述了为分析这两个品系中巨噬细胞的功能活性而建立的模型系统。我们已使来自B10A.Bcgs和B10A.Bcgr近交系小鼠的骨髓巨噬细胞永生化，并分别获得了命名为B10S和B10R的克隆巨噬细胞系。B10R和B10S细胞系表现出巨噬细胞典型的表面标志物和形态。B10S和B10R在吞噬活性、c-fms水平、转化生长因子β（TGFβ）mRNA表达以及对干扰素-γ（IFNγ）加脂多糖（LPS）的反应中肿瘤杀伤活性的表达方面相似。然而，与B10S巨噬细胞相比，用IFNγ激活时，B10R巨噬细胞表达更高水平的Ia mRNA。对这两个细胞系抑菌活性的分析表明，B10R巨噬细胞在抑制耻垢分枝杆菌复制方面比B10S活跃得多。为了测量杆菌的细胞内破坏，设计了一种基于与分枝杆菌核糖体RNA特异性寡核苷酸探针杂交的杀菌试验。结果表明，与B10S相比，B10R巨噬细胞具有增强的组成型杀菌活性。总之，我们从B10A.Bcgs和B10A.Bcgr小鼠的骨髓中获得了巨噬细胞系，它们在相似程度上表达巨噬细胞的功能和表型特征。然而，我们证明，相对于B10S巨噬细胞，B10R巨噬细胞具有更高的Ia mRNA表达，并且它们在表达杀分枝杆菌活性方面组成型更活跃。
