In situ detection of interleukin-1 mRNA in human monocytes.
The interleukin-1 (IL-1) family of soluble pleiotropic immunoregulatory and proinflammatory peptides has at least two distinct members, alpha IL-1 and beta IL-1. Since beta IL-1 is the predominant species in human monocytes, this study was undertaken to identify its mRNA in monocytes using in situ hybridization with a 35S-dCTP labelled beta IL-1 cDNA probe. Grain count analysis demonstrated that adherent lipopolysaccharide-stimulated monocytes were positive, while unstimulated monocytes, lymphocytes and neutrophils, and cells probed with vector only (35S-labelled pBR322) were all negative. We have also shown that in situ hybridization is approx. 13-fold more sensitive than conventional hybridization and in addition this technique allows visualization of mRNA coding for IL-1 in individual cells with morphology preserved. We conclude that in situ hybridization is a specific and sensitive technique for the detection of beta IL-1 mRNA in individual human peripheral blood monocytes.
Tron
V A
VA
Department of Medicine, McMaster University, Ontario, Canada.
Harley
C B
CB
Caussy
D
D
Sauder
D N
DN
D001345:Autoradiography; D004247:DNA; D006801:Humans; D007375:Interleukin-1 / Q000235:genetics*; D008070:Lipopolysaccharides / Q000494:pharmacology; D009000:Monocytes / Q000032:analysis / Q000276:immunology*; D009693:Nucleic Acid Hybridization; D012333:RNA, Messenger / Q000032:analysis*
D007375:Interleukin-1; D008070:Lipopolysaccharides; D012333:RNA, Messenger; D004247:DNA