Bacterial pneumonia is a leading cause of mortality in the United States. Innate immune responses, including type-1 cytokine production, are critical to the effective clearance of bacterial pathogens from the lung. Synthetic oligodeoxynucleotides (ODN) containing unmethylated CpG dinucleotide motifs (CpG ODN), which mimic the effects of bacterial DNA, have been shown to enhance type-1 cytokine responses during infection due to intracellular pathogens, resulting in enhanced microbial clearance. The role of CpG ODN in modulating protective innate immunity against extracellular pathogens is unknown. Using a murine model of Gram-negative pneumonia, we found that CpG ODN administration stimulated protective immunity against Klebsiella pneumoniae. Specifically, intratracheal (it) administration of CpG ODN (30 μg) 48 h before it K. pneumoniae challenge resulted in increased survival, compared with animals pretreated with control ODN or saline. Pretreatment with CpG ODN resulted in enhanced bacterial clearance in lung and blood, and higher numbers of pulmonary neutrophils, NKT cells, γδ-T cells, and activated NK1. 1+ cells and γδ-T lymphocytes during infection. Furthermore, pretreatment with CpG ODN enhanced the production of TNF-α, and type-1 cytokines, including IL-12, IFN-γ, and the IFN-γ-dependent ELR− CXC chemokines IFN-γ-inducible protein-10 and monokine induced by IFN-γ in response to Klebsiella challenge, compared with control mice. These findings indicate that it administration of CpG ODN can stimulate multiple components of innate immunity in the lung, and may form the basis for novel therapies directed at enhancing protective immune responses to severe bacterial infections of the lung.