Pseudomonas aeruginosa adaptation in cystic fibrosis patients increases C5a levels and promotes neutrophil recruitment.

Cystic fibrosis (CF) disease is characterized by an intense airway inflammatory response mediated by neutrophils and chronic respiratory infections caused by P. aeruginosa. High levels of the complement component C5a, the strongest neutrophil chemoattractant molecule, are commonly found in the CF lung and have been associated with a worsening of the disease. In this study, we investigated how the isolates from CF patients modulate the levels of C5a and identified the bacterial factors involved. We demonstrated that most isolates from airway chronic infections induce the production and accumulation of C5a, an effect attributable to the loss of C5a cleavage by the exoproteases alkaline protease (AprA) and elastase B (LasB). Furthermore, we found that lack of the bacterial protease-dependent C5a degradation is due to mutations in the master regulator LasR. Thus, complementation of a non-C5a-cleaving CF isolate with a functional wild-type LasR restored its ability to express both proteases, cleave C5a and reduce neutrophil recruitment in vitro. These findings suggest that the non-cleaving C5a phenotype acquired by the LasR variants frequently isolated in CF patients may account for the strong neutrophilia and general neutrophil dysfunction predisposing toward increased inflammation and reduced bacterial clearance described in CF patients.

Surfactant Protein A Recognizes Outer Membrane Protein OprH on Pseudomonas aeruginosa Isolates From Individuals With Chronic Infection.

Surfactant protein A (SP-A) plays a critical role in the clearance of Pseudomonas aeruginosa from the lung. However, there is limited information about the interaction of this protein with P. aeruginosa isolates from individuals with cystic fibrosis (CF). We characterized the interplay between SP-A and a collection of isogenic sequential isolates from 7 patients with CF. We identified outer membrane protein OprH as a novel ligand for SP-A on P. aeruginosa The last-available (late) isolates from patients with CF bound significantly less SP-A than their respective first-available (early) isolates. This difference could be associated with a reduction in the expression of OprH. Binding of SP-A to OprH promoted phagocytic killing; thus, late CF isolates were at least 2-fold more resistant to SP-A-mediated killing by human macrophages than their respective early isolates. We postulate that the reduction of OprH expression is a previously unrecognized adaptation of P. aeruginosa to the lung of individuals with CF that facilitates the escape of the microorganism from SP-A-mediated phagocytic killing.