Adaptation of Pseudomonas aeruginosa to the chronic phenotype by mutations in the algTmucABD operon in isolates from Brazilian cystic fibrosis patients.

Chronic lung infection by Pseudomonas aeruginosa is the leading cause of morbidity and mortality in cystic fibrosis (CF) patients. This is associated with the conversion of the non-mucoid to the mucoid phenotype. However, there is little information about the occurrence of alginate-producing P. aeruginosa in CF patients outside Europe and North America. The aim of the present study was to investigate mutations in the algTmucABD operon in mucoid and non-mucoid isolates from Brazilian CF patients. Twenty-seven mucoid and 37 non-mucoid isolates from 40 CF patients chronically infected by P. aeruginosa attending a CF reference center in Brazil were evaluated by sequence analysis. Mutations in mucA were observed in 93% of the mucoid isolates and 54% of the non-mucoid isolates. Among these non-mucoid isolates, 55% were considered revertants, since they also had mutations in algT (algU). Most isolates associated with moderate alginate production presented point mutations in mucB and/or mucD. We identified 30 mutations not previously described in the operon. In conclusion, mutations in mucA were the main mechanism of conversion to mucoidy, and most of the non-mucoid isolates were revertants, but the mechanism of revertance is not fully explained by changes in algT.

IgG avidity to Pseudomonas aeruginosa over the course of chronic lung biofilm infection in cystic fibrosis.

BACKGROUND AND OBJECTIVES: The mechanisms leading to low effectiveness of the humoral immune response against P. aeruginosa in cystic fibrosis (CF) are poorly understood. The aim of the present study was to assess the avidity maturation of specific antipseudomonal IgG before and during the development of chronic lung infection in a cohort of Danish CF patients. METHODS: Avidity maturation was assessed against a pooled P. aeruginosa antigen (St-Ag) and against P. aeruginosa alginate in 10 CF patients who developed chronic lung infection and 10 patients who developed intermittent lung colonization, using an ELISA technique with the thiocyanate elution method. Avidity was quantitatively determined by calculating the avidity Constant (Kav). RESULTS: IgG avidity to St-Ag significantly increased at the onset (Median Kav=2.47) and one year after the onset of chronic infection (Median Kav=3.27), but did not significantly changed in patients who developed intermittent colonization. IgG avidity against alginate did not significantly change over the years neither in patients who developed chronic lung infection (Median Kav=3.84 at the onset of chronic infection), nor in patients who developed intermittent colonization. CONCLUSION: IgG avidity to P. aeruginosa alginate does not significantly enhance as chronic lung infection progresses. This probably plays a role in the difficulty to mount an effective opsonophagocytic killing to clear mucoid P. aeruginosa infection in CF.