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Blood, Vol. 95 No. 5 (March 1), 2000:
pp. 1616-1625
A soluble form of human Delta-like-1 inhibits differentiation
of hematopoietic progenitor cells
Wei Han,
Qian Ye, and
Malcolm A. S. Moore
From the Laboratory of Developmental Hematopoiesis, Memorial
Sloan-Kettering Cancer Center, New York, NY.
Two Notch ligand families, Delta and Serrate/Jagged, have been
identified in vertebrates. Members of the Jagged family have been shown
to affect in vitro hematopoiesis. To determine whether members of the
Delta family might play a similar role in hematopoiesis, we examined
the expression of mouse Delta-like-1 (mDll1). mDll1 protein was
detected in whole marrow and in a marrow stromal cell line MS-5. At the
RNA level, both mDll1 and Notch1 were seen in marrow
precursor, differentiated hematopoietic, marrow stromal, and MS-5
cells. We isolated a cDNA encoding the human homologue of mDll1,
designated human Delta-like-1 (hDll1). A soluble form of hDll1, hDll1NDSL, containing the DSL domain and the
N-terminal sequences, was expressed and purified from bacteria as a
glutathione S-transferase (GST) fusion protein. We observed that
hDll1NDSL delayed the acquisition of
differentiation markers by murine hematopoietic progenitor cells
(Lin ) cultured in vitro with cytokines. In addition, it
promoted greater expansion (more than 3 times) of the primitive
hematopoietic precursor cell population, measured in high-proliferative
potential colony assay and day 12 colony-forming unit spleen (CFU-S)
assay, than GST controls. We also observed that the percentage of
apoptotic cells decreased and that the number of cells in the S-phase
of the cell cycle increased in the cultures of Lin
cells with hDll1NDSL. The effects of
hDll1NDSL were blocked by antibody against the mouse
counterpart of hDll1NDSL, mDll1NDSL. These
observations demonstrate that hDll1 plays a role in mediating cell fate
decisions during hematopoiesis.
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