Smith M F, Kueppers F R, Young P R, Lee J C
Department of Immunology and Microbiology, Temple University School of Medicine, Philadelphia, PA 19140.
J Immunol Methods. 1989 Mar 31;118(2):265-72. doi: 10.1016/0022-1759(89)90015-x.
We have used IL-1 alpha and IL-1 beta specific RNA standards in an RNA dot blot assay to rapidly and quantitatively assess the differential expression of the two human interleukin-1 mRNAs. This approach enabled us to minimize inaccuracies due to differences in specific activities of the probes, transcript size, and transfer and/or hybridization efficiencies of the two transcripts. We were thus able to accurately determine the steady state levels and molar ratio of the two species of IL-1 mRNA in human peripheral blood monocytes (PBM) and alveolar macrophages (AM). PBM, upon stimulation by lipopolysaccharide (LPS), expressed approximately ten-fold more IL-1 beta mRNA than IL-1 alpha. AM, however, expressed only two-fold more IL-1 beta then IL-1 alpha. PBM aged in vitro for 7 days prior to LPS stimulation, expressed at least 20-fold more IL-1 beta than IL-1 alpha RNA and at levels significantly lower than either PBM or AM.
我们在RNA斑点印迹分析中使用白细胞介素-1α（IL-1α）和白细胞介素-1β（IL-1β）特异性RNA标准品，以快速、定量地评估两种人类白细胞介素-1 mRNA的差异表达。这种方法使我们能够将由于探针比活性差异、转录本大小以及两种转录本的转移和/或杂交效率差异所导致的误差降至最低。因此，我们能够准确测定人类外周血单核细胞（PBM）和肺泡巨噬细胞（AM）中两种IL-1 mRNA的稳态水平和摩尔比。经脂多糖（LPS）刺激后，PBM表达的IL-1β mRNA比IL-1α多大约10倍。然而，AM表达的IL-1β仅比IL-1α多2倍。在LPS刺激前体外培养7天的PBM，表达的IL-1β RNA比IL-1α至少多20倍，且表达水平明显低于PBM或AM。
