Notch signaling induces cytoplasmic CD3 epsilon expression in human differentiating NK cells.
It has been proposed that heterogeneity in natural killer (NK)-cell phenotype and function can be achieved through distinct thymic and bone marrow pathways of NK-cell development. Here, we show a link between Notch signaling and the generation of intracellular CD3epsilon (cyCD3)-expressing NK cells, a cell population that can be detected in vivo. Differentiation of human CD34(+) cord blood progenitors in IL-15-supplemented fetal thymus organ culture or OP9-Delta-like 1 (DL1) coculture resulted in a high percentage of cyCD3(+) NK cells that was blocked by the gamma-secretase inhibitor DAPT. The requirement for Notch signaling to generate cyCD3(+) NK cells was further illustrated by transduction of CD34(+) cord blood (CB) cells with either the active intracellular part of Notch or the dominant-negative mutant of mastermind-like protein 1 that resulted in the generation of NK cells with respectively high or low frequencies of cyCD3. Human thymic CD34(+) progenitor cells displayed the potential to generate cyCD3(+) NK cells, even in the absence of Notch/DL1 signaling. Peripheral blood NK cells were unable to induce cyCD3 expression after DL1 exposure, indicating that Notch-dependent cyCD3 expression can only be achieved during the early phase of NK-cell differentiation.
De Smedt
Magda
M
Department of Clinical Chemistry, Microbiology, and Immunology, Ghent University Hospital, Ghent, Belgium.
Taghon
Tom
T
Van de Walle
Inge
I
De Smet
Greet
G
Leclercq
Georges
G
Plum
Jean
J
D000818:Animals; D018952:Antigens, CD34 / Q000378:metabolism; D017252:CD3 Complex / Q000378:metabolism*; D019002:CD56 Antigen / Q000378:metabolism; D002135:Calcium-Binding Proteins; D002454:Cell Differentiation; D002465:Cell Movement; D002478:Cells, Cultured; D018920:Coculture Techniques; D003593:Cytoplasm / Q000378:metabolism*; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000378:metabolism; D007694:Killer Cells, Natural / Q000166:cytology / Q000378:metabolism*; D008565:Membrane Proteins / Q000378:metabolism; D051379:Mice; D010641:Phenotype; D051880:Receptors, Notch / Q000378:metabolism*; D015398:Signal Transduction; D013950:Thymus Gland / Q000378:metabolism; D013997:Time Factors
D018952:Antigens, CD34; D017252:CD3 Complex; C115335:CD3E protein, human; D019002:CD56 Antigen; D002135:Calcium-Binding Proteins; C512941:DLK1 protein, human; D036341:Intercellular Signaling Peptides and Proteins; D008565:Membrane Proteins; D051880:Receptors, Notch