Characterization of mononuclear-phagocyte terminal maturation by mRNA phenotyping using a set of cloned cDNA probes.
The terminal step in the maturation of mononuclear cells from circulating monocytes to resident macrophages is accompanied by dramatic changes in cell morphology and physiology. Applying a cultivation system which allows peripheral monocytes to undergo terminal maturation in vitro under absolutely endotoxin-free conditions, we have determined the pattern of expression of a set of eight genes by mRNA phenotyping. The results can be summarized as follows: the two protease inhibitors alpha 1-antitrypsin and alpha 2-macroglobulin show a inverse pattern of expression. alpha 1-Antitrypsin mRNA is repressed, alpha 2-macroglobulin mRNA is strongly induced during maturation to macrophages. Therefore, these two genes are excellent markers of the terminal maturation. In addition, ferritin-light-chain mRNA progressively increases during the course of differentiation, providing a further marker for maturation. Gene expression as a function of activation was studied in mononuclear cells stimulated with bacterial endotoxin (lipopolysaccharide). In monocytes, complement-factor-B, interleukin-1 and interleukin-6 mRNAs are drastically induced upon lipopolysaccharide activation whereas lysozyme RNA is strongly repressed. However, the ability of all four genes to respond to endotoxin was markedly diminished or abolished in mature macrophages, indicating that susceptibility to a certain type of activation may be restricted to a specific stage of maturation. Our data show that mRNA phenotyping is excellently suited for the characterization of the differentiation and activation state of mononuclear phagocytes.
Ganter
U
U
Medizinische Universitätsklinik, Freiburg.
Bauer
J
J
Majello
B
B
Gerok
W
W
Ciliberto
G
G
D015152:Blotting, Northern; D002454:Cell Differentiation; D002478:Cells, Cultured; D015342:DNA Probes; D005786:Gene Expression Regulation / Q000187:drug effects; D005796:Genes; D006801:Humans; D008070:Lipopolysaccharides / Q000494:pharmacology; D008264:Macrophages / Q000166:cytology*; D009000:Monocytes / Q000166:cytology*; D010586:Phagocytes / Q000166:cytology*; D010641:Phenotype; D012333:RNA, Messenger / Q000235:genetics*; D014158:Transcription, Genetic
D015342:DNA Probes; D008070:Lipopolysaccharides; D012333:RNA, Messenger