The respiratory tract contains numerous antimicrobial factors necessary for normal innate pulmonary defense. Although many of these molecules reside in airway surface liquid (ASL) simultaneously, little information exists concerning antagonistic, additive, or synergistic interactions. Since both cationic lysozyme and anionic antimicrobial peptides (AP) are found in high concentrations in ASL, the purpose of this study was to assess any interaction that might affect antimicrobial activity. For this, Pasteurella haemolytica, Micrococcus lysodeikticus, or Pseudomonas aeruginosa were added to egg white lysozyme (3.9-250.0 microg/ml) or human neutrophil lysozyme (0.8-50.0 microg/ml) and H-GADDDDD-OH (from 0.01 to 0.50 mM) mixtures in 50, 100, or 150 mM NaCl; incubated for 2 h; and then plated. In this assay, the MICs of AP for P. haemolytica, M. lysodeikticus, and P. aeruginosa varied slightly depending upon the concentration of NaCl and MICs generally increased slightly with increasing NaCl concentrations. The MIC of lysozyme for P. haemolytica and M. lysodeikticus also increased slightly with increasing NaCl concentrations. The MIC of lysozyme for P. aeruginosa was greater than 50 microg/ml and did not vary with increasing NaCl concentrations. When AP was combined with lysozyme in 50, 100, or 150 mM NaCl concentrations, there was no significant interaction that affected antimicrobial activity. In conclusion, the MICs of AP generally increased with increasing NaCl concentrations but lysozyme and AP appeared not to interact significantly at physiologically relevant concentrations.