Notch activation induces the generation of functional NK cells from human cord blood CD34-positive cells devoid of IL-15.
The development of NK cells from hematopoietic stem cells is thought to be dependent on IL-15. In this study, we demonstrate that stimulation of human cord blood CD34(+) cells by a Notch ligand, Delta4, along with IL-7, stem cell factor, and Fms-like tyrosine kinase 3 ligand, but no IL-15, in a stroma-free culture induced the generation of cells with characteristics of functional NK cells, including CD56 and CD161 Ag expression, IFN-gamma secretion, and cytotoxic activity against K562 and Jurkat cells. Addition of gamma-secretase inhibitor and anti-human Notch1 Ab to the culture medium almost completely blocked NK cell emergence. Addition of anti-human IL-15-neutralizing Ab did not affect NK cell development in these culture conditions. The presence of IL-15, however, augmented cytotoxicity and was required for a more mature NK cell phenotype. CD56(+) cells generated by culture with IL-15, but without Notch stimulation, were negative for CD7 and cytoplasmic CD3, whereas CD56(+) cells generated by culture with both Delta4 and IL-15 were CD7(+) and cytoplasmic CD3(+) from the beginning and therefore more similar to in vivo human NK cell progenitors. Together, these results suggest that Notch signaling is important for the physiologic development of NK cells at differentiation stages beyond those previously postulated.
Department of Cell Therapy and Transplantation Medicine, University of Tokyo Hospital, Tokyo, Japan.
Kyoko
K
Haraguchi
Takahiro
T
Suzuki
Noriko
N
Koyama
Keiki
K
Kumano
Fumio
F
Nakahara
Akihiko
A
Matsumoto
Yasuhisa
Y
Yokoyama
Mamiko
M
Sakata-Yanagimoto
Shigeo
S
Masuda
Tsuyoshi
T
Takahashi
Aki
A
Kamijo
Koki
K
Takahashi
Minoko
M
Takanashi
Yoshiki
Y
Okuyama
Koji
K
Yasutomo
Seiji
S
Sakano
Hideo
H
Yagita
Mineo
M
Kurokawa
Seishi
S
Ogawa
Shigeru
S
Chiba
D018952:Antigens, CD34 / Q000276:immunology / Q000378:metabolism; D002454:Cell Differentiation / Q000276:immunology*; D005312:Fetal Blood; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000276:immunology; D006801:Humans; D019409:Interleukin-15 / Q000276:immunology / Q000378:metabolism*; D007694:Killer Cells, Natural / Q000166:cytology* / Q000276:immunology; D016131:Lymphocyte Subsets / Q000166:cytology* / Q000276:immunology; D051880:Receptors, Notch / Q000276:immunology / Q000378:metabolism*; D015398:Signal Transduction / Q000276:immunology
D018952:Antigens, CD34; D019409:Interleukin-15; D051880:Receptors, Notch