The Na+/Ca2+ exchanger (NCX) is a plasma membrane protein that can switch Na+ and Ca2+ in either direction to maintain the homeostasis of intracellular Ca2+. A family of three genes (NCX1, NCX2, and NCX3) has been identified in neurons and muscle cells. NCX activity has also been reported in certain immune cells (e.g., mast cells). We have examined the expression and function of these NCX isoforms in the human monocytes and lung macrophages. Monocytes were purified from peripheral blood of healthy donors. Macrophages (HLM) were isolated and purified from the lung parenchyma of patients undergoing thoracic surgery. NCX1 and NCX3, but not NCX2, were expressed in HLM and monocytes at both mRNA and protein level. Exposure to Na+-free medium induced a significant increase in intracellular calcium concentration ([Ca2+]i) in both cell types, suggesting that NCX isoforms expressed on these cells were functionally active. This response was completely abolished by the NCX inhibitor 5-(N-4-chlorobenzyl) -20,40-dimethylbenzamil (CB-DMB). In addition, incubation of macrophages with Na+-free medium induced a marked release of TNF-α. Preincubation of HLM with CB-DMB and RNAi-mediated knockdown of NCX1 blocked TNF-α release. Our results demonstrate that human macrophages and monocytes express NCX1 and NCX3 that operate in a bidirectional manner to restore [Ca2+]i to generate Ca2+ signals and to induce TNF-α production. We suggest that NCX may modulate Ca2+ homeostasis and proinflammatory functions in human macrophages and monocytes.