title: NK cells differentiated from bone marrow, cord blood and peripheral blood stem cells exhibit similar phenotype and functions.
abstract: In the present study, we investigated the differentiation of human NK cells from bone marrow, cord blood and mobilized peripheral blood purified CD34+ stem cells using a potent culture system. Elutriated CD34+ stem cells were grown for several weeks in medium supplemented with stem cell factor (SCF) and IL-15 in the presence or absence of a murine stromal cell line (MS-5). Our data indicate that IL-15 induced the proliferation and maturation of highly positive CD56+ NK cells in both types of culture, although murine stromal cells slightly increased the proliferation of NK cells. NK cells differentiated in the presence of MS-5 were mostly CD56+ CD7 and a small subset expressed CD16. These in vitro differentiated CD56+ NK cells displayed cytolytic activity against the HLA class I- target K562. The CD56+ CD16+ subset also lysed NK-resistant Daudi cells. Neither of these NK subsets were shown to express Fas ligand. Total CD56+ cells expressed high amounts of transforming growth factor-beta and granulocyte-macrophage colony-stimulating factor, but no IFN-gamma. Investigation of NK receptor expression showed that most CD56+ cells expressed membrane CD94 and NKG2-A mRNA. PCR analysis revealed that p58 was also expressed in these cells. The role of CD94 in NK cell-mediated cytotoxicity was assessed on human HLA-B7-transfected murine L cells. While a low cytotoxic activity towards HLA-B7 cells was observed, the HLA-DR4 control cells were killed with high efficiency. These studies demonstrate that cytolytic and cytokine-producing NK cells may be derived from adult and fetal precursors by IL-15 and that these cells express a CD94 receptor which may influence their lytic potential.
authors.affiliation: INSERM U 487, Institut Gustave Roussy, Villejuif, France.
authors.forename: G
authors.initials: G
authors.lastname: Carayol
authors.forename: C
authors.initials: C
authors.lastname: Robin
authors.forename: J H
authors.initials: JH
authors.lastname: Bourhis
authors.forename: A
authors.initials: A
authors.lastname: Bennaceur-Griscelli
authors.forename: S
authors.initials: S
authors.lastname: Chouaib
authors.forename: L
authors.initials: L
authors.lastname: Coulombel
authors.forename: A
authors.initials: A
authors.lastname: Caignard
mesh_terms: D036541:ADP-ribosyl Cyclase; D051997:ADP-ribosyl Cyclase 1; D000328:Adult; D015703:Antigens, CD / Q000096:biosynthesis / Q000502:physiology; D018952:Antigens, CD34 / Q000276:immunology; D019011:Antigens, CD7 / Q000276:immunology; D000943:Antigens, Differentiation / Q000276:immunology; D001854:Bone Marrow Cells / Q000166:cytology; D019002:CD56 Antigen / Q000276:immunology; D003601:Cytotoxicity Tests, Immunologic; D053222:Fas Ligand Protein; D005312:Fetal Blood / Q000166:cytology; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000096:biosynthesis; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000276:immunology; D006801:Humans; D016130:Immunophenotyping; D007371:Interferon-gamma / Q000235:genetics; D019409:Interleukin-15 / Q000494:pharmacology; D007694:Killer Cells, Natural / Q000166:cytology / Q000276:immunology / Q000502:physiology*; D037181:Lectins, C-Type; D019891:Leukopoiesis; D008233:Lymphotoxin-alpha / Q000096:biosynthesis; D008562:Membrane Glycoproteins / Q000096:biosynthesis / Q000276:immunology / Q000502:physiology; D009244:NAD+ Nucleosidase / Q000276:immunology; D051920:NK Cell Lectin-Like Receptor Subfamily D; D012333:RNA, Messenger; D017452:Receptors, IgG / Q000276:immunology; D016212:Transforming Growth Factor beta / Q000096:biosynthesis; D014407:Tumor Cells, Cultured
chemical_list: D015703:Antigens, CD; D018952:Antigens, CD34; D019011:Antigens, CD7; D000943:Antigens, Differentiation; D019002:CD56 Antigen; C505661:FASLG protein, human; D053222:Fas Ligand Protein; D019409:Interleukin-15; C497869:KLRD1 protein, human; D037181:Lectins, C-Type; D008233:Lymphotoxin-alpha; D008562:Membrane Glycoproteins; D051920:NK Cell Lectin-Like Receptor Subfamily D; D012333:RNA, Messenger; D017452:Receptors, IgG; D016212:Transforming Growth Factor beta; D007371:Interferon-gamma; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D036541:ADP-ribosyl Cyclase; C498154:CD38 protein, human; D009244:NAD+ Nucleosidase; D051997:ADP-ribosyl Cyclase 1