A rapid and quantitative method for the determination of interleukin-1 alpha and -beta mRNA expression in human monocytes and macrophages.
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.
Smith
M F
MF
Department of Immunology and Microbiology, Temple University School of Medicine, Philadelphia, PA 19140.
Kueppers
F R
FR
Young
P R
PR
Lee
J C
JC
D015152:Blotting, Northern / Q000379:methods; D002478:Cells, Cultured; D006801:Humans; D007375:Interleukin-1 / Q000032:analysis* / Q000235:genetics; D008264:Macrophages / Q000032:analysis* / Q000378:metabolism; D009000:Monocytes / Q000032:analysis* / Q000378:metabolism; D009693:Nucleic Acid Hybridization; D012333:RNA, Messenger / Q000032:analysis* / Q000096:biosynthesis; D014158:Transcription, Genetic
D007375:Interleukin-1; D012333:RNA, Messenger