Oxygen availability affects cell differentiation, survival and function, with profound consequences on tissue homeostasis, inflammation and immunity. A gradient of oxygen levels is present in most organs of the body as well as in virtually every site of inflammation, damaged or pathological tissue. As a consequence, infiltrating leukocytes, macrophages in particular, are equipped with the capacity to shift their metabolism to anaerobic glycolysis, to generate ATP and induce the expression of factors that increase the supply of oxygen and nutrients. Strikingly, low oxygen conditions (hypoxia) and inflammatory signals share selected transcriptional events, including the activation of members of both the hypoxia-inducible factor and nuclear factor κB families, which may converge to activate specific cell programs. In the pathological response to hypoxia, cancer in particular, macrophages act as orchestrators of disease evolution and their number can be used as a prognostic marker. Here we review mechanisms of macrophage adaptation to hypoxia, their role in disease as well as new perspectives for their therapeutic targeting.