title: Surfactant dysfunction and lung injury due to the E. coli virulence factor hemolysin in a rat pneumonia model.
abstract: This study tests the hypothesis that the virulence factor hemolysin (Hly) expressed by extraintestinal pathogenic Escherichia coli contributes to surfactant dysfunction and lung injury in a rat model of gram-negative pneumonia. Rats were instilled intratracheally with CP9 (wild type, Hly-positive), CP9hlyA (Hly-minus), CP9/pEK50 (supraphysiological Hly), or purified LPS. At 6 h postinfection, rats given CP9 had a decreased percentage content of large surfactant aggregates in cell-free bronchoalveolar lavage (BAL), decreased large aggregate surface activity, decreased Pa(O2)/FiO2) ratio, increased BAL albumin/protein levels, and increased histological evidence of lung injury compared with rats given CP9hlyA or LPS. In addition, rats given CP9/pEK50 or CP9 had decreased large aggregate surface activity, decreased Pa(O2)/FiO2) ratios, and increased BAL albumin/protein levels at 2 h postinfection compared with rats given CP9hlyA. The severity of permeability lung injury based on albumin/protein levels in BAL at 2 h was ordered as CP9/pEK50 > CP9 > CP9hlyA > normal saline controls. Total lung titers of bacteria were increased at 6 h in rats given CP9 vs. CP9hlyA, but bacterial titers were not significantly different at 2 h, indicating that increased surfactant dysfunction and lung injury were associated with Hly as opposed to bacterial numbers per se. Further studies in vitro showed that CP9 could directly lyse transformed pulmonary epithelial cells (H441 cells) but that indirect lysis of H441 cells secondary to Hly-induced neutrophil lysis did not occur. Together, these data demonstrate that Hly is an important direct mediator of surfactant dysfunction and lung injury in gram-negative pneumonia.
authors.affiliation: Department of Medicine, The Witebsky Center for Microbial Pathogenesis, NY, USA. trusso@acsu.buffalo.edu
authors.forename: Thomas A
authors.initials: TA
authors.lastname: Russo
authors.forename: Zhengdong
authors.initials: Z
authors.lastname: Wang
authors.forename: Bruce A
authors.initials: BA
authors.lastname: Davidson
authors.forename: Stacy A
authors.initials: SA
authors.lastname: Genagon
authors.forename: Janet M
authors.initials: JM
authors.lastname: Beanan
authors.forename: Ruth
authors.initials: R
authors.lastname: Olson
authors.forename: Bruce A
authors.initials: BA
authors.lastname: Holm
authors.forename: Paul R
authors.initials: PR
authors.lastname: Knight
authors.forename: Patricia R
authors.initials: PR
authors.lastname: Chess
authors.forename: Robert H
authors.initials: RH
authors.lastname: Notter
mesh_terms: D000818:Animals; D001427:Bacterial Toxins / Q000494:pharmacology*; D018893:Bronchoalveolar Lavage; D004195:Disease Models, Animal; D029968:Escherichia coli Proteins / Q000494:pharmacology*; D006460:Hemolysin Proteins / Q000494:pharmacology*; D008168:Lung / Q000187:drug effects / Q000378:metabolism*; D055370:Lung Injury; D009504:Neutrophils / Q000473:pathology; D018410:Pneumonia, Bacterial / Q000378:metabolism* / Q000473:pathology; D011663:Pulmonary Surfactants / Q000378:metabolism*; D051381:Rats; D020318:Rats, Long-Evans
chemical_list: D001427:Bacterial Toxins; D029968:Escherichia coli Proteins; D006460:Hemolysin Proteins; C036507:Hlya protein, E coli; D011663:Pulmonary Surfactants