RAPID COMMUNICATION

Interleukin-1 and Tumor Necrosis Factor-, But Not

Interleukin-6, Stimulate Osteoprotegerin Ligand Gene
Expression in Human Osteoblastic Cells
L. C. HOFBAUER,
1
D. L. LACEY,
2
C. R. DUNSTAN,
2
T. C. SPELSBERG,
3
B. L. RIGGS,
1
and
S. KHOSLA
1
1
Endocrine Research Unit and
3
Department of Biochemistry and Molecular Biology, Mayo Clinic and Mayo Foundation, Rochester, MN, USA
2
Amgen Inc., Thousand Oaks, CA, USA
Recent studies have identified osteoprotegerin ligand
(OPG-L) as the essential factor required for osteoclastogen-
esis, and that the effects are prevented by its soluble receptor,
osteoprotegerin (OPG). However, there are limited data at
present on the regulation of OPG-L expression in human
Key Words: Interleukin (IL)-1; IL-6; Osteoblast; Osteoprote-
gerin; Osteoprotegerin ligand; Tumor necrosis factor-


IL-6 soluble receptor (IL-6sR) were from R&D Systems (Min-
neapolis, MN).
Cell Cultures
The following human osteoblastic cells were used: (i) a condi-
tionally immortalized bipotential marrow stromal cell line
(hMS)
9
; (ii) normal marrow stromal cells (MS) obtained by bone
Discussion
The balance between bone resorption and bone formation is
regulated by an integrated network of cytokines that includes
macrophage colony-stimulating factor (M-CSF), IL-1, IL-4,
IL-6, IL-11, prostaglandin estradiol (PGE
2
), TNF-, and trans-
forming growth factor- (TGF-), which are synthesized by
bone marrow stromal cells and osteoblasts and which modulate
osteoclast differentiation and activity.
15,22,25
Although there is
man osteoblastic lineage cells. Thus, the effect of these cytokines
in stimulating osteoclastogenesis may depend on the ratio of
OPG-L:OPG generated in the bone microenvironment. A similar
situation appears to exist in the case of other cytokines and their
endogenous antagonists, including IL-1, soluble IL-1 receptor,
IL-1 receptor antagonist,
6,33
and matrix metalloproteinases and
tissue inhibitor of metalloproteinases.
2
Regulation of the ratio
agonist:antagonist may represent another level of regulation in
these complex systems. In the case of the OPG-L/OPG system, it
appears that TNF- and IL-1 induce parallel, rather than recip-
rocal, changes of both components. It should be noted, however,
that while mRNA levels of both OPG-L and OPG increased
following stimulation by TNF- and IL-1, the net effect of
these changes on osteoclastogenesis clearly depends on the
biological dose responses to these agents as well as on other
issues such as relative changes in the respective protein concen-
trations. Clearly, further studies are needed to address these
issues.
In conclusion, we find that IL-1 and TNF-, but not IL-6,
stimulate steady-state mRNA levels of OPG-L, which is a critical
factor for osteoclastogenesis in various human osteoblastic lin-
eage cells. These data suggest that an increase in OPG-L pro-
duction following exposure of osteoblastic lineage cells to IL-1
and TNF- may provide an important paracrine mechanism
whereby these proinflammatory cytokines stimulate osteoclasto-
gensesis, thus promoting bone resorption and bone loss in vivo.
Acknowledgments: The authors acknowledge the technical assistance of
M. J. Schroeder, B. Ngo, and R. A. Soderberg. This work was supported
by Grant AG-04875 from the National Institutes of Health. Dr. Hofbauer
is a recipient of a postdoctoral fellowship from the Deutsche Forschungs-
gemeinschaft (Ho 1875/1-1).
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