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. 2017 Oct 26;7:14159. doi: 10.1038/s41598-017-12761-1

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© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Panel A reports column chart of % IBV/SV ratio in control mice, CLL untreated mice (CLL no treatment) and CLL Denosumab ones. White columns indicate bone erosion at CT2 (four weeks after CLL cells administration) while grey columns report the same variable at CT3. AT CT2, CLL was associated with a significant femur erosion (*p < 0.01 vs controls). This difference persisted in untreated mice at CT3 (three weeks later). By contrast, Denosumab induced a significant reduction in IBV/SV to values that were lower with respect to untreated mice and similar to ones of control group. Panel B, displays the individual response to treatment (blue lines) or placebo (red lines) from CT2 to CT3. Denosumab caused a significant reduction of IBV/SV. Panel C summarizes the effect of in vivo denosumab treatment on human neoplastic B-cells (h CD45/CD5/CD19- positive cells). Percentage of CLL cells was significantly lower in denosumab treated mice compared to controls treated with saline (CTR) in BM and spleen. The mean and SEM of 19 mice injected with CLL cells from two different cases are shown (Mann-Whitney test). Panel D displays the direct correlation between %RANK⁺ B cells (X axis) in peripheral blood of each patient and actively proliferating cells indexed by HDF (Y axis). Panel E: displays the in vitro reduction in HDF induced by Denosumab. Panel F: displays the direct relationship between %RANK⁺ cells (X axis) and Denosumab effect on HDF.