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and Phosphatidylinositol 3'-Kinase and Promotion of Macrophage Differentiation by Insulin-Like Growth Factor-I1
*
Laboratory of Immunophysiology, Department of Animal Sciences, and
College of Medicine, Department of Pathology, University of Illinois, Urbana, IL 61801; and
Institut National de la Recherche Agronomique-INSERM, Unité de Recherches de Neurobiologie des Comportements, Bordeaux, France
Phosphoinositides that are phosphorylated at the D3 position have
been reported to activate an atypical,
Ca2-independent protein kinase C (PKC) isoform
designated PKC-
, and overexpression of this enzyme leads to
monocytic differentiation. In this study, we cultured human HL-60
promyeloid cells with vitamin D3 and insulin-like growth
factor-I (IGF-I), a 70-amino-acid peptide that activates
phosphatidylinositol 3'-kinase (PI 3-kinase) in murine promyeloid
cells. Two days later, the proportion of cells differentiating into
macrophages in serum-free medium, as assessed by expression of the
-subunit of the ß2 integrin CD11b, increased from
5 ± 1% to 25 ± 3%. Addition of IGF-I increased the
proportion of cells differentiating into CD11b-positive macrophages to
78 ± 5%. In the absence of vitamin D3, IGF-I did not
induce expression of CD11b (6 ± 1%). The IGF-I-promoted
macrophage differentiation was blocked specifically by preincubation of
HL-60 cells with a mAb (IR3) directed against the IGF type I
receptor. Similarly, pretreatment of cells with either IR3 or an
IGF-binding protein, IGFBP-3, led to a 75% inhibition of CD11b
expression when cells were cultured with vitamin D3 in
serum-containing medium. IGF-I, but not vitamin D3, caused
a sevenfold increase in the enzymatic activity of both PI 3-kinase and
atypical PKC-. Inhibition of IGF-I-inducible PI 3-kinase with either
wortmannin or LY294002 abrogated the IGF-I-induced activation of
PKC- and totally blocked the enhancement in macrophage
differentiation caused by IGF-I. These data establish that PKC- is a
putative downstream target of PI 3-kinase that is activated during
IGF-I-promoted macrophage differentiation.
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