Differential Expression of Receptors for Interleukin-3 on Subsets of CD34-Expressing Hematopoietic Cells of Rhesus Monkeys

• Published in: Blood 1995-07, Vol.86 (2), p.581-591
• Main Authors: Wognum, Albertus W., Visser, Trudy P., Jong, Marg O. de, Egeland, Torstein, Wagemaker, Gerard
• Format: Article
• Language: English
• Subjects: Animals; Antigens, CD - analysis; Antigens, CD20; Antigens, CD34; Antigens, Differentiation, B-Lymphocyte - analysis; Biological and medical sciences; Blood Cells - drug effects; Blood Cells - metabolism; Bone Marrow Cells; CD2 Antigens - analysis; Cell Differentiation; Cell differentiation, maturation, development, hematopoiesis; Cell physiology; Cells, Cultured; Drug Synergism; Fundamental and applied biological sciences. Psychology; Granulocyte-Macrophage Colony-Stimulating Factor - pharmacology; Hematopoiesis - drug effects; Hematopoietic Cell Growth Factors - pharmacology; Hematopoietic Stem Cells - drug effects; Hematopoietic Stem Cells - metabolism; Histocompatibility Antigens Class II - analysis; Immunophenotyping; Interleukin-3 - pharmacology; Interleukin-6 - pharmacology; Macaca mulatta - metabolism; Molecular and cellular biology; Neprilysin - analysis; Receptors, Interleukin-3 - biosynthesis; Receptors, Interleukin-3 - genetics; Stem Cell Factor; Tumor Necrosis Factor Receptor Superfamily, Member 7 - analysis
• ISSN: 0006-4971; 1528-0020
• DOI: 10.1182/blood.V86.2.581.bloodjournal862581

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 CD34dullRhLA-DRbrightCD10+/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 dono-genie erythroid and myeloid progenitors (burst-forming unit-erythroid and colony-forming unit-culture), whereas CD34brightIL-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 fin