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Effect of bone morphogenetic protein‐6 on macrophages

Bone morphogenetic proteins (BMPs) are members of the transforming growth factor (TGF)‐β superfamily which regulates bone formation, haematopoiesis and development. While TGF‐β is known to be a negative regulator of the immune system, the effect of BMPs on the immune system is largely unknown. Herein, the effect of BMP‐6 on the innate immune system was investigated using the murine macrophage cell line RAW 264·7. BMP‐6 altered cellular morphology, inhibited cellular proliferation, increased the fraction of subG1 phase cells, and decreased the fraction of cells in the S and G2M phases, without changing the percentage of apoptotic cells. In addition, BMP‐6 induced expression of pro‐inflammatory inducible nitric oxide synthase (iNOS) and the cytokine tumour necrosis factor (TNF)‐α. Reverse transcription–polymerase chain reaction (RT‐PCR) analysis demonstrated the expression of all three known type II BMP receptors [BMP‐RII, activin (Act)‐RIIA and Act‐RIIB] and two of the three known type I receptors [activin receptor‐like kinase 2 (ALK2) and ALK3]. Over‐expression as well as knock‐down studies using short hairpin RNA (shRNA) demonstrated that BMP‐RII, ALK2 and ALK3 are the functional BMP‐6 receptors in macrophages. Finally, the effect of BMP‐6 was confirmed in murine peritoneal macrophages and the THP‐1 human monocyte cell line. Taken together, these results demonstrate that BMP‐6 regulates the proliferation and gene expression profile of macrophages.