muramidase and Pneumonia--Bacterial

Lysozyme, a 14-kDa protein, is one of the most abundant antimicrobials in the lungs. Its concentration in airway surface sufficient to kill several bacterial pathogens in vitro. The purpose of this study was to determine if administration of exogenous lysozyme would further enhance bacterial killing in vivo.. To assess the effect of acute lung infection on endogenous lysozyme protein levels, mice were infected by intratracheal instillation of Pseudomonas aeruginosa and bronchoalveolar (BAL) fluid assessed for lysozyme concentration and for muramidase activity. In order to inform in vivo testing, species-specific bacterial killing efficacy was determined by incubating mucoid P. aeruginosa with 2×10. These results indicate that endogenous lysozyme is increased during acute lung infection and that early administration of exogenous lysozyme further enhances bacterial killing in vivo.

Topics: Animals; Anti-Infective Agents; Bronchoalveolar Lavage Fluid; Drug Synergism; Drug Therapy, Combination; Female; Lung; Male; Mice; Microbial Viability; Muramidase; Pneumonia, Bacterial; Pseudomonas aeruginosa; Pseudomonas Infections; Treatment Outcome

Resident immune cells (e.g., macrophages [MΦs]) and airway mucus clearance both contribute to a healthy lung environment. To investigate interactions between pulmonary MΦ function and defective mucus clearance, a genetic model of lysozyme M (LysM) promoter-mediated MΦ depletion was generated, characterized, and crossed with the sodium channel β subunit transgenic (Scnn1b-Tg) mouse model of defective mucus clearance. Diphtheria toxin A-mediated depletion of LysM(+) pulmonary MΦs in wild-type mice with normal mucus clearance resulted in lethal pneumonia in 24% of neonates. The pneumonias were dominated by Pasteurella pneumotropica and accompanied by emaciation, neutrophilic inflammation, and elevated Th1 cytokines. The incidence of emaciation and pneumonia reached 51% when LysM(+) MΦ depletion was superimposed on the airway mucus clearance defect of Scnn1b-Tg mice. In LysM(+) MΦ-depleted Scnn1b-Tg mice, pneumonias were associated with a broader spectrum of bacterial species and a significant reduction in airway mucus plugging. Bacterial burden (CFUs) was comparable between Scnn1b-Tg and nonpneumonic LysM(+) MΦ-depleted Scnn1b-Tg mice. However, the nonpneumonic LysM(+) MΦ-depleted Scnn1b-Tg mice exhibited increased airway inflammation, the presence of neutrophilic infiltration, and increased levels of inflammatory cytokines in bronchoalveolar lavage fluid compared with Scnn1b-Tg mice. Collectively, these data identify key MΦ-mucus clearance interactions with respect to both infectious and inflammatory components of muco-obstructive lung disease.

Topics: Animals; Animals, Newborn; Cytokines; Diphtheria Toxin; Disease Models, Animal; Epithelial Sodium Channels; Genetic Predisposition to Disease; Inflammation Mediators; Luminescent Proteins; Lung; Macrophages; Mice, Inbred C57BL; Mice, Transgenic; Mucociliary Clearance; Muramidase; Pasteurella Infections; Pasteurella pneumotropica; Peptide Fragments; Phenotype; Pneumonia, Bacterial; Promoter Regions, Genetic

Keratinocyte growth factor (KGF) is an epithelial mitogen that has been reported to protect the lungs from a variety of toxic and infectious insults. In prior studies we found that recombinant human KGF accelerates clearance of bacteria from the murine lung by augmenting the function of alveolar macrophages (AM). In this study we tested the hypothesis that endogenous KGF plays a role in the maintenance of innate pulmonary defense against gram-negative bacterial infections. KGF-deficient mice exhibited delayed clearance of Escherichia coli from the lungs, attenuated phagocytosis by AM, and decreased antimicrobial activity in bronchoalveolar lavage (BAL) fluid, due in part to reductions in levels of surfactant protein A, surfactant protein D, and lysozyme. These immune deficits were accompanied by lower alveolar type II epithelial cell counts and reduced alveolar type II epithelial cell expression of collectin and lysozyme genes on a per cell basis. No significant between-group differences were detected in selected inflammatory cytokines or BAL inflammatory cell populations at baseline or after bacterial challenge in the wild-type and KGF-deficient mice. A single intranasal dose of recombinant human KGF reversed defects in bacterial clearance, AM function, and BAL fluid antimicrobial activity. We conclude that KGF supports alveolar innate immune defense through maintenance of alveolar antimicrobial protein levels and functions of AM. Together these data demonstrate a role for endogenous KGF in maintenance of normal pulmonary innate immune function.

Topics: Animals; Cells, Cultured; Collectins; Escherichia coli Infections; Female; Fibroblast Growth Factor 7; Gene Expression; Humans; Immunity, Innate; Macrophages, Alveolar; Mice, 129 Strain; Mice, Inbred C57BL; Mice, Knockout; Muramidase; Pneumonia, Bacterial; Pulmonary Alveoli

Previous studies from this laboratory have shown that aerosolized recombinant human lysozyme (rhLZ) mitigates Pseudomonas aeruginosa (PA)-induced pneumonia. In the current investigation, our laboratory tested the hypothesis that aerosolized rhLZ can potentiate the effects of tobramycin (TBMN), thereby reducing the effective dose of this agent in the treatment of PA-induced pneumonia. Syrian hamsters were instilled intratracheally with PA, then exposed to an aerosol containing either 1% rhLZ, 3 μg TBMN, or a combination of both agents. In contrast to the initial studies with rhLZ, which involved 3 separate aerosol exposures, only a single treatment was used in the current investigation. Twenty-four hours after completion of the aerosol regimen, the following parameters were measured: (1) whole-lung colony-forming units (CFU), (2) total bronchoalveolar lavage fluid (BALF) CFU, (3) lung histopathology, and (4) total BALF neutrophils. The combination of rhLZ and TBMN significantly reduced whole-lung and BALF CFU, as well as the inflammatory index, compared to TBMN alone. Similar results were seen in vitro with regard to bactericidal activity. These findings provide a rationale for clinical testing of rhLZ as an adjunct to commercial antibiotic treatment.

Topics: Administration, Inhalation; Animals; Anti-Bacterial Agents; Bronchoalveolar Lavage Fluid; Cricetinae; Disease Models, Animal; Drug Synergism; Female; Humans; Lung; Mesocricetus; Muramidase; Neutrophils; Pneumonia, Bacterial; Pseudomonas aeruginosa; Pseudomonas Infections; Recombinant Proteins; Tobramycin

As an alternative to conventional antibiotics, aerosolized recombinant human lysozyme (rhLZ) was used to treat experimentally induced pneumonia. Syrian hamsters were inoculated intratracheally with a nonmucoid strain of Pseudomonas aeruginosa (PA), then exposed to a 1.0% solution of rhLZ in water for 2 hours per day for 3 consecutive days (controls were treated with aerosolized water alone). Compared to controls, the rhLZ-treated group showed statistically significant reductions in the following parameters: (1) lung histopathological changes, (2) bacterial colony-forming units in whole lung and bronchoalveolar lavage fluid (BALF), (3) total BALF leukocytes, (4) percent BALF neutrophils, and (5) alveolar septal apoptosis. Exposure to aerosolized rhLZ also resulted in a large increase in BALF lysozyme activity. These findings indicate that aerosolized rhLZ may be potentially useful in reducing the level of bacterial colonization and inflammation in the lungs of patients with PA pneumonia.

Topics: Administration, Inhalation; Animals; Anti-Infective Agents; Apoptosis; Bronchoalveolar Lavage Fluid; Cricetinae; Drug Evaluation, Preclinical; Female; Humans; Leukocyte Count; Lung; Mesocricetus; Muramidase; Pneumonia, Bacterial; Pseudomonas aeruginosa; Pseudomonas Infections; Recombinant Proteins

To determine the antimicrobial activity of nisin F against Staphylococcus aureus in the respiratory tract.. The respiratory tract of nonimmunosuppressed and immunosuppressed Wistar rats were colonized with 4 x 10(5) viable cells of S. aureus K and then treated by administering 8192 arbitrary units (AU) nisin F intranasal. Symptoms of pneumonia were detected in the trachea and lungs of immunosuppressed rats that had not been treated with nisin F. The trachea and lungs of immunosuppressed rats treated with nisin F were healthy. No significant differences were recorded in blood cell indices. The antimicrobial activity of low concentrations nisin F (80-320 AU ml(-1)) was slightly stimulated by lysozyme and lactoferrin.. Nisin F inhibited the growth of S. aureus K in the respiratory tract of immunocompromised rats. Treatment with nisin F at 8192 AU proofed safe, as the trachea, lungs, bronchi and haematology of the rats appeared normal.. Nisin F is nontoxic and may be used to control respiratory tract infections caused by S. aureus. This is, however, a preliminary study with an animal model and need to be confirmed with studies on humans.

Topics: Administration, Intranasal; Animals; Bronchi; Drug Interactions; Humans; Immunocompromised Host; Lactoferrin; Lung; Male; Muramidase; Nisin; Pneumonia, Bacterial; Rats; Rats, Wistar; Respiratory Tract Infections; Staphylococcal Infections; Staphylococcus aureus; Trachea