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Blood, Vol. 92 No. 3 (August 1), 1998:
pp. 901-907
Negative Regulation by Interleukin-3 (IL-3) of Mouse Early B-Cell
Progenitors and Stem Cells in Culture: Transduction of the Negative
Signals by  c and IL-3 Proteins of IL-3 Receptor and Absence of
Negative Regulation by Granulocyte-Macrophage Colony-Stimulating
Factor
Takuya Matsunaga,
Fumiya Hirayama,
Yuji Yonemura,
Richard Murray, and
Makio Ogawa
From the Department of Veterans Affairs Medical Center and the
Department of Medicine, Medical University of South Carolina,
Charleston, SC; and DNAX Research Institute of Molecular and Cellular
Biology, Palo Alto, CA.
The receptors for interleukin-3 (IL-3), granulocyte-macrophage
colony-stimulating factor (GM-CSF), and IL-5 share a common signaling
subunit  c. However, in the mouse, there is an additional IL-3
signaling protein, IL-3, which is specific for IL-3. We have
previously reported that IL-3 abrogates the lymphoid potentials of
murine lymphohematopoietic progenitors and the reconstituting ability
of hematopoietic stem cells. We used bone marrow cells from c- and
IL-3-knock-out mice to examine the relative contributions of the
receptor proteins to the negative regulation by IL-3. First, we tested
the effects of IL-3 on lymphohematopoietic progenitors by using
lineage-negative (Lin ) marrow cells of 5-fluorouracil
(5-FU)-treated mice in the two-step methylcellulose culture we reported
previously. Addition of IL-3 to the combination of steel factor (SF,
c-kit ligand) and IL-11 abrogated the B-lymphoid potential of the
marrow cells of both types of knock-out mice as well as wild-type mice.
Next, we investigated the effects of IL-3 on in vitro expansion of the
hematopoietic stem cells. We cultured
LinSca-1-positive, c-kit-positive marrow cells from
5-FU-treated mice in suspension in the presence of SF and IL-11 with
or without IL-3 for 7 days and tested the reconstituting ability of the
cultured cells by transplanting the cells into lethally irradiated Ly-5 congenic mice together with "compromised" marrow cells. Presence of IL-3 in culture abrogated the reconstituting ability of the cells
from both types of knock-out mice and the wild-type mice. In contrast,
addition of GM-CSF to the suspension culture abrogated neither B-cell
potential nor reconstituting abilities of the cultured cells of
wild-type mice. These observations may have implications in the choice
of cytokines for use in in vitro expansion of human hematopoietic stem
cells and progenitors.
© 1998 by The American Society of Hematology.
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