Beck Rose C, Padival Mallika, Yeh David, Ralston Justine, Cooke Kenneth R, Lowe John B
Department of Pathology, Case Western Reserve University School of Medicine, Cleveland, Ohio, USA.
Biol Blood Marrow Transplant. 2009 Sep;15(9):1026-37. doi: 10.1016/j.bbmt.2009.06.002.
Notch receptor signaling is required for T cell development, but its role in natural killer (NK) cell development is poorly understood. We compared the ability of the 5 mammalian Notch ligands (Jagged1, Jagged2, Delta1, Delta3, or Delta4) to induce NK cell development from human hematopoietic progenitor cells (HPCs). CD34(+) HPCs were cultured with OP9 stromal cell lines transduced with 1 of the Notch ligands or with OP9 stromal cells alone, in the presence of IL-7, Flt3L, and IL-15. Differentiation and expansion of CD56(+)CD3(-) cells were greatly accelerated in the presence of Jagged2, Delta-1, or Delta-4, versus culture in the absence of ligand or in the presence of Jagged1 or Delta3. At 4 weeks, cultures containing Jagged2, Delta1, or Delta4 contained 80% to 90% NK cells, with the remaining cells being CD33(+) myelogenous cells. Notch-induced NK (N-NK) cells resembled CD56(bright) NK cells in that they were CD16(-), CD94(-), CD117(+), and killer immunoglobulin-like receptors (KIR(-)). They also expressed NKp30, NKp44, NKp46, 2B4, and DNAM-1, with partial expression of NKG2D. The N-NK cells displayed cytotoxic activity against the K562 and RPMI-8226 cell lines, at levels similar to activated peripheral blood (PB) NK cells, although killing of Daudi cells was not present. N-NK cells were also capable of interferon (IFN)-gamma secretion. Thus, Notch ligands have differential ability to induce and expand immature, but functional, NK cells from CD34(+) HPCs. The use of Notch ligands to generate functional NK cells in vitro may be significant for cellular therapy purposes.
Notch受体信号传导对于T细胞发育是必需的，但其在自然杀伤（NK）细胞发育中的作用却知之甚少。我们比较了5种哺乳动物Notch配体（Jagged1、Jagged2、Delta1、Delta3或Delta4）诱导人造血祖细胞（HPC）发育为NK细胞的能力。将CD34(+) HPC与转导了1种Notch配体的OP9基质细胞系或单独的OP9基质细胞一起培养，同时添加IL-7、Flt3L和IL-15。与无配体培养或存在Jagged1或Delta3的培养相比，存在Jagged2、Delta-1或Delta-4时，CD56(+)CD3(-)细胞的分化和扩增显著加速。在4周时，含有Jagged2、Delta1或Delta4的培养物中含有80%至90%的NK细胞，其余细胞为CD33(+)骨髓细胞。Notch诱导的NK（N-NK）细胞类似于CD56(bright) NK细胞，因为它们是CD16(-)、CD94(-)、CD117(+)，且杀伤性免疫球蛋白样受体（KIR(-)）。它们还表达NKp30、NKp44、NKp46、2B4和DNAM-1，NKG2D部分表达。N-NK细胞对K562和RPMI-8226细胞系具有细胞毒性活性，其水平与活化的外周血（PB）NK细胞相似，尽管对Daudi细胞无杀伤作用。N-NK细胞也能够分泌干扰素（IFN）-γ。因此，Notch配体具有不同的能力来诱导和扩增来自CD34(+) HPC的未成熟但有功能的NK细胞。利用Notch配体在体外产生功能性NK细胞对于细胞治疗目的可能具有重要意义。
