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Published online before print September 2, 2004

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(Journal of Leukocyte Biology. 2004;76:1214-1219.)
© 2004 by Society for Leukocyte Biology

G-CSF-mobilized CD34+ cells cultured in interleukin-2 and stem cell factor generate a phenotypically novel monocyte

Stem Cell Allotransplantation Section, Hematology Branch, National Heart Lung and Blood Institute, National Institutes of Health, Bethesda, Maryland

¹ Correspondence: Hematology Branch, Hematopoietic Stem Cell Transplantation Section, National Heart Lung and Blood Institute, National Institutes of Health, 9000 Rockville Pike, Bethesda MD 20892-0001. E-mail: giuseppe.sconocchia{at}roswellpark.org

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 CD56^dim, mainly CD33+ (2.5±2, n=11) cells with macrophage morphology. The CD56^bright population had cytoplasmic granzyme A but lacked killer inhibitory receptor, suggesting they were immature NK cells. The CD56^dim, 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, CD56^dimCD33+ 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-. 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.

Key Words: NK cells • monocytes/macrophages • cellular proliferation • cellular differentiation

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