Burkholderia mallei is a gram-negative bacterial pathogen of domestic equidae and humans that can cause severe, rapidly life-threatening pneumonic infections. Little is known regarding the role of chemokines and early cellular immune responses in protective immunity to pulmonary infection with B. mallei. Although the role of MCP-1 in gram-positive bacterial infections has been previously investigated, the role of MCP-1 in immunity to acute pneumonia caused by gram-negative bacteria, such as B. mallei, has not been assessed. In a mouse model of pneumonic B. mallei infection, we found that both MCP-1(-/-) mice and CCR2(-/-) mice were extremely susceptible to pulmonary infection with B. mallei, compared with wild-type (WT) C57Bl/6 mice. Bacterial burden and organ lesions were significantly increased in CCR2(-/-) mice, compared with WT animals, following B. mallei challenge. Monocyte and dendritic cell recruitment into the lungs of CCR2(-/-) mice was significantly reduced in comparison with that in WT mice following B. mallei infection, whereas neutrophil recruitment was actually increased. Depletion of monocytes and macrophages prior to infection also greatly raised the susceptibility of WT mice to infection. Production of IL-12 and IFN-gamma in the lungs after B. mallei infection was significantly impaired in both MCP-1(-/-) and CCR2(-/-) mice, whereas treatment of CCR2(-/-) mice with rIFN-gamma restored protection against lethal challenge with B. mallei. Thus, we conclude that MCP-1 plays a key role in regulating cellular immunity and IFN-gamma production following pneumonic infection with B. mallei and therefore may also figure importantly in other gram-negative pneumonias.