Nitrogen-containing bisphosphonates (N-BPs) are front-line treatments for preventing bone loss in patients with osteoporosis and metastatic bone disease. N-BPs are generally considered to act only in the skeleton due to their high affinity for calcium ions in bone mineral. These drugs exert anti-resorptive actions by disabling bone-degrading osteoclasts via inhibition of the mevalonate pathway. Surprisingly however, recent studies suggest that N-BPs also have beneficial, pleiotropic effects outside the skeleton, including a lower risk of pneumonia infection/mortality, but the mechanisms remain unknown. Like osteoclasts, macrophages in culture and in tumours in vivo efficiently internalise N-BPs by endocytosis. Thus, we sought to investigate whether BPs could also act directly on macrophages in the lung in vivo and potentially alter immune function.
We used fluorescently-labelled N-BP and flow cytometry to detect drug uptake into cells. 24 hours after iv injection, we detected N-BP in >98% of CD11blo/-CD11chiF4/80+ alveolar macrophages in lung, as well as 80% of peritoneal cells (almost entirely CD11b+F4/80+ large peritoneal macrophages). Characteristic features of N-BP action – inhibition of protein prenylation and build-up of the metabolite isopentenyl pyrophosphate – were also detected in alveolar and peritoneal macrophages 48 hours after one iv dose of the N-BP zoledronate. Importantly, systemic zoledronate treatment significantly enhanced the production of proinflammatory cytokines and chemokines in lung by 2.5-5-fold (including IL-1beta, IL-6, TNFalpha, CXCL1, CCL2,3,4,5) upon intranasal endotoxin challenge.
In summary, we present new evidence that N-BP treatment targets tissue resident macrophages in vivo in lung and peritoneum and enhances immune responses, dispelling the long-held dogma that these drugs act only in bone. Given that inhibition of the mevalonate pathway is known to have a variety of anti-microbial effects, we propose that the apparent beneficial effects of N-BP therapy on pneumonia infection and mortality occur by boosting the immune response of alveolar macrophages to infection.