Differential expression of receptors for interleukin-3 on subsets of CD34-expressing hematopoietic cells of rhesus monkeys.
The target cell specificity of interleukin-3 (IL-3) was examined by flow cytometric analysis of IL-3 receptor (IL-3R) expression on rhesus monkey bone marrow (BM) cells using biotinylated IL-3. Only 2% to 5% of unfractionated cells stained specifically with the biotinylated IL-3 and most of these cells were present within the CD34+ subset. IL-3Rs were detected on small CD34dull/RhLA-DRbright/CD10+/CD27+/CD2-/++ +CD20- cells, which probably represent B-cell precursors. IL-3R+ CD34- BM cells, which were detected at low frequencies, consisted of small CD20dull/surface-IgM+/RhLA-DR+ cells. These cells represented immature B lymphocytes, whereas CD20bright mature B cells were IL-3R-. The highest IL-3R levels were detected on CD34dull/RhLA-DRbright blast-like cells. These cells differentiated into monocytes, neutrophils, and basophils after IL-3 and/or granulocyte-macrophage colony-stimulating factor (GM-CSF) stimulation in vitro. The CD34bright/IL-3R- subset contained all clonogenic erythroid and myeloid progenitors (burst-forming unit-erythroid and colony-forming unit-culture), whereas CD34bright/IL-3Rdull cells differentiated into monocytes, neutrophils, and erythroid cells after shorter culture periods. This finding showed that IL-3R expression increases during monocyte and granulocyte differentiation. Results of three-color experiments indicated that IL-3Rs are expressed on CD34bright/RhLA-DRbright cells as well as on CD34bright/RhLA-DRdull cells, with the latter population expression approximately twofold to threefold lower IL-3R levels. A large fraction (> 30%) of single-cell/well-sorted CD34bright/RhLA-DRdull cells formed multilineage colonies after 2 to 4 weeks of stimulation with IL-3, GM-CSF, Kit ligand, and IL-6. Individual colonies contained cells that still expressed CD34 as well as differentiated monocytes, granulocytes, and erythroid cells. These results confirmed that the CD34bright/RhLA-DRdull subset was enriched for immature, multipotent progenitor cells, whereas the CD34bright/RhLA-DRbright population mainly contained lineage-committed precursors. The results are consistent with the concept that IL-3Rs are induced at very early stages of hematopoiesis, as identified by high expression of CD34 and low expression of RhLA-DR. IL-3R expression continues to be low during differentiation into lineage-committed progenitors; gradually increases on differentiating progenitor cells for B cells, granulocytes, monocytes, and, possibly also, erythrocytes; but finally declines to undetectable levels during terminal differentiation into mature cells of all lineages in peripheral blood, with the exception of basophils.(ABSTRACT TRUNCATED AT 400 WORDS)
Wognum
A W
AW
Department of Hematology, Erasmus Universiteit, Rotterdam, The Netherlands.
Visser
T P
TP
de Jong
M O
MO
Egeland
T
T
Wagemaker
G
G
D000818:Animals; D015703:Antigens, CD / Q000032:analysis*; D018951:Antigens, CD20; D018952:Antigens, CD34; D000944:Antigens, Differentiation, B-Lymphocyte / Q000032:analysis; D001773:Blood Cells / Q000187:drug effects / Q000378:metabolism; D001854:Bone Marrow Cells; D018801:CD2 Antigens / Q000032:analysis; D002454:Cell Differentiation; D002478:Cells, Cultured; D004357:Drug Synergism; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000494:pharmacology; D006410:Hematopoiesis / Q000187:drug effects; D016298:Hematopoietic Cell Growth Factors / Q000494:pharmacology; D006412:Hematopoietic Stem Cells / Q000187:drug effects / Q000378:metabolism*; D000949:Histocompatibility Antigens Class II / Q000032:analysis; D016130:Immunophenotyping; D007377:Interleukin-3 / Q000494:pharmacology; D015850:Interleukin-6 / Q000494:pharmacology; D008253:Macaca mulatta / Q000378:metabolism*; D015260:Neprilysin / Q000032:analysis; D016185:Receptors, Interleukin-3 / Q000096:biosynthesis* / Q000235:genetics; D019089:Stem Cell Factor; D018127:Tumor Necrosis Factor Receptor Superfamily, Member 7 / Q000032:analysis
D015703:Antigens, CD; D018951:Antigens, CD20; D018952:Antigens, CD34; D000944:Antigens, Differentiation, B-Lymphocyte; D018801:CD2 Antigens; D016298:Hematopoietic Cell Growth Factors; D000949:Histocompatibility Antigens Class II; D007377:Interleukin-3; D015850:Interleukin-6; D016185:Receptors, Interleukin-3; D019089:Stem Cell Factor; D018127:Tumor Necrosis Factor Receptor Superfamily, Member 7; C094588:antigen RhLA-DR; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D015260:Neprilysin