Fig. 5. IL-10R inhibition reverse TAM-induced MM proliferation.

A Effect of macrophages on MM proliferation. MM.1S-CBR-GFP cells were cultured alone or co-cultured at 1:1 ratio with THP-1-derived macrophages for 3 or 7 days in 3DTEBM. MM proliferation is determined by GFP cell count, presented as % of MM alone. Statistical significance is compared between the co-culture condition and the respective monoculture condition of each day. B Effect of IL-10R inhibition on TAM-induced MM proliferation and cell cycle signaling. THP-1-derived macrophages were pretreated with or without of anti-IL-10R mAb (5µg/ml) for 4 h, and MM.1S cells were cultured alone or with macrophages at 1:1 ratio for 24 h. MM.1S cells were then detached from macrophages, lyzed, and proteins were analyzed by western for proliferation (pAKT and S6R) and cell cycle signaling (pRB). Actin was blotted as the loading control. C Effect of IL10R inhibition on TAM-induced MM proliferation. MM.1S-CBR-GFP cells were cultured alone or co-cultured at 1:1 ratio with THP-1-derived macrophages in 3DTEBM and treated with or without anti-IL-10R mAb (5 µg/ml) for 3 days. MM survival is determined by GFP cell count by flow cytometry, presented as % of MM monoculture NT condition. Statistical significance is compared between anti-IL-10R mAb and NT. D Effect of anti-IL-10R mAb on in vivo MM tumor progression. NSG mice (n = 14) were inoculated with MM.1S-CBR-GFP and injected with human PBMC-derived macrophages. Treatment started at day 14 post-inoculation, mice were randomly divided into two groups which were treated with vehicle control (n = 7), or anti-IL-10R mAb (5 mg/kg, i.p, twice/week, n = 7). Tumor progression was measured at days 14, 28, and 42 by BLI, and represented as fold of day 14. Statistical significance is compared between anti-IL-10R mAb and NT. (Bars = Average ± SD, **p < 0.01; ***p < 0.001).