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St. Vincent’s Institute of Medical Research, Fitzroy, Victoria, Australia
IL-12, like IL-18, was shown to potently inhibit osteoclast
formation in cultures of cocultures of murine osteoblast and spleen
cells, as well as in adult spleen cells treated with M-CSF and receptor
activator of NF-
B ligand (RANKL). Neither IL-12 nor IL-18 was able
to inhibit RANKL-induced osteoclast formation in cultured RAW264.7
cells, demonstrating that IL-12, like IL-18, was unable to act directly
on osteoclastic precursors. IL-12, like IL-18, was found to act by T
cells, since depletion of T cells from the adult spleen cell cultures
ablated the inhibitory action of IL-12 and addition of either CD4 or
CD8 T cells from C57BL/6 mice to RANKL-stimulated RAW264.7 cultures
permitted IL-12 or IL-18 to be inhibitory. Additionally, IL-12 was
still able to inhibit osteoclast formation in cocultures with
osteoblasts and spleen cells from either GM-CSF R-/- mice
or IFN-
R-/- mice, indicating that neither GM-CSF nor
IFN- was mediating osteoclast inhibition in these cultures.
Combined, IL-18 and IL-12 synergistically inhibited osteoclast
formation at concentrations 20- to 1000-fold less, respectively, than
when added individually. A candidate inhibitor could not be
demonstrated using neutralizing Abs to IL-4, IL-10, or IL-13 or from
mRNA expression profiles among known cytokine inhibitors of
osteoclastogenesis in response to IL-12 and IL-18 treatment, although
the unknown inhibitory molecule was determined to be secreted from T
cells.
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