Bacterial lipopolysaccharide (LPS) evokes several functional responses in the neutrophil that contribute to innate immunity. Although certain responses, such as adhesion and synthesis of tumor necrosis factor-alpha, are inhibited by pretreatment with an inhibitor of p38 mitogen-activated protein kinase, others, such as actin assembly, are unaffected. The aim of the present study was to investigate the changes in neutrophil gene transcription and protein expression following lipopolysaccharide exposure and to establish their dependence on p38 signaling. Microarray analysis indicated expression of 13% of the 7070 Affymetrix gene set in nonstimulated neutrophils, and LPS up-regulation of 100 distinct genes, including cytokines and chemokines, signaling molecules, and regulators of transcription. Proteomic analysis yielded a separate list of up-regulated modulators of inflammation, signaling molecules, and cytoskeletal proteins. Poor concordance between mRNA transcript and protein expression changes was noted. Pretreatment with the p38 inhibitor SB203580 attenuated 23% of LPS-regulated genes and 18% of LPS-regulated proteins by > or = 40%. This study indicates that p38 plays a selective role in regulation of neutrophil transcripts and proteins following lipopolysaccharide exposure, clarifies that several of the effects of lipopolysaccharide are post-transcriptional and post-translational, and identifies several proteins not previously reported to be involved in the innate immune response.