G-CSF-mobilized CD34+ cells cultured in interleukin-2 and stem cell factor generate a phenotypically novel monocyte.
To study the early stages of development from stem cells of the CD56+ cell population [which includes natural killer (NK) cells], granulocyte-colony stimulating factor-mobilized peripheral blood CD34+ cells from healthy donors were sorted to >99% purity and cultured in the presence of stem cell factor and interleukin (IL)-2. After 3 weeks in culture, the majority of cells acquired CD33, with or without human leukocyte antigen-DR and CD14. In 20 stem cell donors tested, 8.7 +/- 8.8% of cells were CD56+. Two major CD56+ subsets were identified: CD56(bright), mainly CD33- cells (7+/-10%, n=11) with large, granular lymphocyte morphology, and CD56dim, mainly CD33+ (2.5+/-2, n=11) cells with macrophage morphology. The CD56bright population had cytoplasmic granzyme A but lacked killer inhibitory receptor, suggesting they were immature NK cells. The CD56dim, CD33+, population lacked NK markers. They may represent a minor subset of normal monocytes at a developmental stage comparable with the rare CD56+ CD33+ hybrid myeloid/NK cell leukemia. Consistent with a monocyte nature, CD56dimCD33+ proliferated and produced a variety of cytokines upon lipopolysaccharide stimulation, including IL-8, IL-6, monocyte chemoattractant protein-1, and macrophage-derived chemokine but not interferon-gamma. In a short-term cytotoxicity assay, they failed to kill but powerfully inhibited the proliferation of the NK-resistant cell line P815. The generation of CD56+ cells was negatively regulated by hyaluronic acid and IL-4, indicating that extracellular matrix may play an important role in the commitment of CD34+ cells into CD56 myeloid and lymphoid lineages.
Sconocchia
Giuseppe
G
Hematopoietic Stem Cell Transplantation Section, National Heart Lung and Blood Institute, National Institutes of Health, 9000 Rockville Pike, Bethesda MD 20892-0001, USA. giuseppe.sconocchia@roswellpark.org
Fujiwara
Hiroshi
H
Rezvani
Katayoun
K
Keyvanfar
Keyvan
K
El Ouriaghli
Frank
F
Grube
Matthias
M
Melenhorst
Jos
J
Hensel
Nancy
N
Barrett
A John
AJ
D015703:Antigens, CD / Q000276:immunology; D018952:Antigens, CD34 / Q000276:immunology*; D015214:Antigens, Differentiation, Myelomonocytic / Q000276:immunology; D015415:Biomarkers; D019002:CD56 Antigen / Q000276:immunology*; D002454:Cell Differentiation / Q000187:drug effects / Q000276:immunology*; D019070:Cell Lineage / Q000187:drug effects / Q000276:immunology; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D016207:Cytokines / Q000096:biosynthesis; D003601:Cytotoxicity Tests, Immunologic; D005109:Extracellular Matrix / Q000276:immunology / Q000378:metabolism; D016179:Granulocyte Colony-Stimulating Factor / Q000494:pharmacology; D053804:Granzymes; D006801:Humans; D006820:Hyaluronic Acid / Q000378:metabolism / Q000494:pharmacology; D016130:Immunophenotyping; D007376:Interleukin-2 / Q000494:pharmacology; D007694:Killer Cells, Natural / Q000166:cytology / Q000187:drug effects / Q000276:immunology*; D008213:Lymphocyte Activation / Q000187:drug effects / Q000276:immunology; D008214:Lymphocytes / Q000166:cytology / Q000187:drug effects / Q000276:immunology; D008264:Macrophages / Q000166:cytology / Q000187:drug effects / Q000276:immunology; D009000:Monocytes / Q000166:cytology / Q000187:drug effects / Q000276:immunology*; D023461:Myeloid Progenitor Cells / Q000166:cytology / Q000187:drug effects / Q000276:immunology*; D012697:Serine Endopeptidases / Q000378:metabolism; D063268:Sialic Acid Binding Ig-like Lectin 3; D019089:Stem Cell Factor / Q000494:pharmacology
D015703:Antigens, CD; D018952:Antigens, CD34; D015214:Antigens, Differentiation, Myelomonocytic; D015415:Biomarkers; C571163:CD33 protein, human; D019002:CD56 Antigen; D016207:Cytokines; D007376:Interleukin-2; D063268:Sialic Acid Binding Ig-like Lectin 3; D019089:Stem Cell Factor; D016179:Granulocyte Colony-Stimulating Factor; D006820:Hyaluronic Acid; D053804:Granzymes; D012697:Serine Endopeptidases; C509212:GZMA protein, human