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Soluble TRAIL does not impair the anti-osteoclastic activity of osteoprotegerin.

Zauli G, Rimondi E, Secchiero P - J. Cell. Mol. Med. (2008)

View Article: PubMed Central - PubMed

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Dear Editor: It has been extensively documented that osteoprotegerin (OPG), a soluble member of the TNF receptor superfamily, inhibits osteoclastogenesis by binding to receptor activator of NF-kB ligand (RANKL) and preventing interaction with its cognate transmembrane receptor RANK [reviewed in ]... Interestingly, previous data from different groups have shown that recombinant TRAIL modulates the differentiation of erythroid and myeloid precursors, while inhibits both human and mouse osteoclastogenesis when added to pre-osteoclast cultures induced to differentiate with recombinant macrophage colony-stimulating factor (M-CSF) +RANKL as well as to mature osteoclasts... In this respect, it has been clearly shown that mouse pre-osteoclasts only express TRAIL-R2, while human peripheral blood-derived pre-osteoclasts express both death receptors TRAIL-R1 and TRAIL-R2 as well as TRAIL-R4... Therefore, to further elucidate the important issue of the interplay between RANKL, TRAIL and OPG in human osteoclastogenesis, we have cultured adherent PBMC with M-CSF+RANKL for 14 days in the absence or presence of recombinant OPG and recombinant TRAIL, prepared as previously described... TRAIL and OPG were added alone or in combination... Importantly, all cytokines were used at the same concentration (50 ng/ml)... As expected, the addition of TRAIL to M-CSF+RANKL significantly (P < 0.05) inhibited osteoclast formation (Fig. 1A and B)... Of note, recombinant OPG completely abrograted (P < 0.05) osteoclast formation irrespectively of the presence of recombinant TRAIL in culture (Fig. 1A and B)... The anti-osteoclastic activity of OPG could not be ascribed to a low affinity of OPG for TRAIL since OPG (50 ng/ml) efficiently inhibited the apoptosis induced by TRAIL (50 ng/ml) in HL-60 leukemic cells (data not shown)... Our current observations on one hand confirm that TRAIL has anti-osteoclastic activity and on the other hand indicate that it does not affect the potent anti-osteoclastic activity of OPG at least in the simplified model of osteoclastogenesis represented by human PBMC induced to differentiate by M-CSF+RANKL.

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Effect of combined treatment of RANKL, TRAIL and OPG on osteoclastic differentiation. Adherent PBMC were cultured with M-CSF alone for 6 days. Then cells were cultured for additional 14 days with the addition of RANKL in the absence or presence of TRAIL and/or OPG, as indicated. Cultures were analysed for osteoclastic differentiation by scoring the number of TRAP+ multinucleated cells (A), and measuring the levels of TRAP 5b (B), a specific marker of resorption activity, in culture supernatants by ELISA. Data represent the means ±SD of three different experiments performed in duplicate.
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fig01: Effect of combined treatment of RANKL, TRAIL and OPG on osteoclastic differentiation. Adherent PBMC were cultured with M-CSF alone for 6 days. Then cells were cultured for additional 14 days with the addition of RANKL in the absence or presence of TRAIL and/or OPG, as indicated. Cultures were analysed for osteoclastic differentiation by scoring the number of TRAP+ multinucleated cells (A), and measuring the levels of TRAP 5b (B), a specific marker of resorption activity, in culture supernatants by ELISA. Data represent the means ±SD of three different experiments performed in duplicate.

Mentions: Therefore, to further elucidate the important issue of the interplay between RANKL, TRAIL and OPG in human osteoclastogenesis, we have cultured adherent PBMC with M-CSF+RANKL for 14 days in the absence or presence of recombinant OPG and recombinant TRAIL, prepared as previously described [16]. TRAIL and OPG were added alone or in combination. Importantly, all cytokines were used at the same concentration (50 ng/ml). As expected [11–14], the addition of TRAIL to M-CSF+RANKL significantly (P < 0.05) inhibited osteoclast formation (Fig. 1A and B). Of note, recombinant OPG completely abrograted (P < 0.05) osteoclast formation irrespectively of the presence of recombinant TRAIL in culture (Fig. 1A and B). The anti-osteoclastic activity of OPG could not be ascribed to a low affinity of OPG for TRAIL since OPG (50 ng/ml) efficiently inhibited the apoptosis induced by TRAIL (50 ng/ml) in HL-60 leukemic cells (data not shown).


Soluble TRAIL does not impair the anti-osteoclastic activity of osteoprotegerin.

Zauli G, Rimondi E, Secchiero P - J. Cell. Mol. Med. (2008)

Effect of combined treatment of RANKL, TRAIL and OPG on osteoclastic differentiation. Adherent PBMC were cultured with M-CSF alone for 6 days. Then cells were cultured for additional 14 days with the addition of RANKL in the absence or presence of TRAIL and/or OPG, as indicated. Cultures were analysed for osteoclastic differentiation by scoring the number of TRAP+ multinucleated cells (A), and measuring the levels of TRAP 5b (B), a specific marker of resorption activity, in culture supernatants by ELISA. Data represent the means ±SD of three different experiments performed in duplicate.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4401137&req=5

fig01: Effect of combined treatment of RANKL, TRAIL and OPG on osteoclastic differentiation. Adherent PBMC were cultured with M-CSF alone for 6 days. Then cells were cultured for additional 14 days with the addition of RANKL in the absence or presence of TRAIL and/or OPG, as indicated. Cultures were analysed for osteoclastic differentiation by scoring the number of TRAP+ multinucleated cells (A), and measuring the levels of TRAP 5b (B), a specific marker of resorption activity, in culture supernatants by ELISA. Data represent the means ±SD of three different experiments performed in duplicate.
Mentions: Therefore, to further elucidate the important issue of the interplay between RANKL, TRAIL and OPG in human osteoclastogenesis, we have cultured adherent PBMC with M-CSF+RANKL for 14 days in the absence or presence of recombinant OPG and recombinant TRAIL, prepared as previously described [16]. TRAIL and OPG were added alone or in combination. Importantly, all cytokines were used at the same concentration (50 ng/ml). As expected [11–14], the addition of TRAIL to M-CSF+RANKL significantly (P < 0.05) inhibited osteoclast formation (Fig. 1A and B). Of note, recombinant OPG completely abrograted (P < 0.05) osteoclast formation irrespectively of the presence of recombinant TRAIL in culture (Fig. 1A and B). The anti-osteoclastic activity of OPG could not be ascribed to a low affinity of OPG for TRAIL since OPG (50 ng/ml) efficiently inhibited the apoptosis induced by TRAIL (50 ng/ml) in HL-60 leukemic cells (data not shown).

View Article: PubMed Central - PubMed

AUTOMATICALLY GENERATED EXCERPT
Please rate it.

Dear Editor: It has been extensively documented that osteoprotegerin (OPG), a soluble member of the TNF receptor superfamily, inhibits osteoclastogenesis by binding to receptor activator of NF-kB ligand (RANKL) and preventing interaction with its cognate transmembrane receptor RANK [reviewed in ]... Interestingly, previous data from different groups have shown that recombinant TRAIL modulates the differentiation of erythroid and myeloid precursors, while inhibits both human and mouse osteoclastogenesis when added to pre-osteoclast cultures induced to differentiate with recombinant macrophage colony-stimulating factor (M-CSF) +RANKL as well as to mature osteoclasts... In this respect, it has been clearly shown that mouse pre-osteoclasts only express TRAIL-R2, while human peripheral blood-derived pre-osteoclasts express both death receptors TRAIL-R1 and TRAIL-R2 as well as TRAIL-R4... Therefore, to further elucidate the important issue of the interplay between RANKL, TRAIL and OPG in human osteoclastogenesis, we have cultured adherent PBMC with M-CSF+RANKL for 14 days in the absence or presence of recombinant OPG and recombinant TRAIL, prepared as previously described... TRAIL and OPG were added alone or in combination... Importantly, all cytokines were used at the same concentration (50 ng/ml)... As expected, the addition of TRAIL to M-CSF+RANKL significantly (P < 0.05) inhibited osteoclast formation (Fig. 1A and B)... Of note, recombinant OPG completely abrograted (P < 0.05) osteoclast formation irrespectively of the presence of recombinant TRAIL in culture (Fig. 1A and B)... The anti-osteoclastic activity of OPG could not be ascribed to a low affinity of OPG for TRAIL since OPG (50 ng/ml) efficiently inhibited the apoptosis induced by TRAIL (50 ng/ml) in HL-60 leukemic cells (data not shown)... Our current observations on one hand confirm that TRAIL has anti-osteoclastic activity and on the other hand indicate that it does not affect the potent anti-osteoclastic activity of OPG at least in the simplified model of osteoclastogenesis represented by human PBMC induced to differentiate by M-CSF+RANKL.

Show MeSH
Related in: MedlinePlus