Differentiation of NK1.1+, Ly49+ NK cells from flt3+ multipotent marrow progenitor cells.
To delineate factors involved in NK cell development, we established an in vitro system in which lineage marker (Lin)-, c-kit+, Sca2+ bone marrow cells differentiate into lytic NK1.1+ but Ly49- cells upon culture in IL-7, stem cell factor (SCF), and flt3 ligand (flt3L), followed by IL-15 alone. A comparison of the ability of IL-7, SCF, and flt3L to generate IL-15-responsive precursors suggested that NK progenitors express the receptor for flt3L. In support of this, when Lin-, c-kit+, flt3+ or Lin-, c-kit+, flt3- progenitors were utilized, 3-fold more NK cells arose from the flt3+ than from the flt3- progenitors. Furthermore, NK cells that arose from flt3- progenitors showed an immature NK1.1dim, CD2-, c-kit+ phenotype as compared with the more mature NK1.1bright, CD2+/-, c-kit- phenotype displayed by NK cells derived from flt3+ progenitors. Both progenitors, however, gave rise to NK cells that were Ly49 negative. To test the hypothesis that additional marrow-derived signals are necessary for Ly49 expression on developing NK cells, flt3+ progenitors were grown in IL-7, SCF, and flt3L followed by culture with IL-15 and a marrow-derived stromal cell line. Expression of Ly49 molecules, including those of which the MHC class I ligands were expressed on the stromal or progenitor cells, as well as others of which the known ligands were absent, was induced within 6-13 days. Thus, we have established an in vitro system in which Ly49 expression on developing NK cells can be analyzed and possibly experimentally manipulated.
Williams
N S
NS
Department of Pathology, Graduate Program in Immunology, University of Texas Southwestern Medical Center, Dallas 75235, USA. williams.n@pathology.swmed.edu
Klem
J
J
Puzanov
I J
IJ
Sivakumar
P V
PV
Bennett
M
M
Kumar
V
V
D000818:Animals; D000941:Antigens / Q000276:immunology*; D000950:Antigens, Ly; D000954:Antigens, Surface; D001854:Bone Marrow Cells / Q000166:cytology* / Q000201:enzymology / Q000276:immunology; D002454:Cell Differentiation / Q000276:immunology; D019070:Cell Lineage / Q000276:immunology; D002478:Cells, Cultured; D018920:Coculture Techniques; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000201:enzymology / Q000276:immunology; D019409:Interleukin-15 / Q000502:physiology; D007694:Killer Cells, Natural / Q000166:cytology* / Q000201:enzymology / Q000276:immunology; D037181:Lectins, C-Type; D008024:Ligands; D016131:Lymphocyte Subsets / Q000166:cytology* / Q000201:enzymology / Q000276:immunology; D008562:Membrane Glycoproteins / Q000096:biosynthesis*; D008565:Membrane Proteins / Q000096:biosynthesis / Q000494:pharmacology; D051379:Mice; D008810:Mice, Inbred C57BL; D008811:Mice, Inbred DBA; D016513:Mice, SCID; D055651:NK Cell Lectin-Like Receptor Subfamily B; D011506:Proteins / Q000276:immunology*; D011518:Proto-Oncogene Proteins / Q000096:biosynthesis*; D019009:Proto-Oncogene Proteins c-kit / Q000096:biosynthesis; D020794:Receptor Protein-Tyrosine Kinases / Q000096:biosynthesis*; D055630:Receptors, NK Cell Lectin-Like; D017154:Stromal Cells / Q000166:cytology / Q000276:immunology; D051941:fms-Like Tyrosine Kinase 3
D000941:Antigens; D000950:Antigens, Ly; D000954:Antigens, Surface; D019409:Interleukin-15; C527211:Klrb1c protein, mouse; D037181:Lectins, C-Type; D008024:Ligands; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D055651:NK Cell Lectin-Like Receptor Subfamily B; D011506:Proteins; D011518:Proto-Oncogene Proteins; D055630:Receptors, NK Cell Lectin-Like; C084881:flt3 ligand protein; C084839:thymic shared antigen-1; C497971:Flt3 protein, mouse; D019009:Proto-Oncogene Proteins c-kit; D020794:Receptor Protein-Tyrosine Kinases; D051941:fms-Like Tyrosine Kinase 3