File S1 - Characterization of Nontypable Haemophilus influenzae Isolates Recovered from Adult Patients with Underlying Chronic Lung Disease Reveals Genotypic and Phenotypic Traits Associated with Persistent Infection

This file contains Figures S1 to S5. Figure S1. Characterization of serial clonal isolates recovered from patient 1, NTHi1a, 1b and 1c. (A) Analysis of Haemophilus influenzae lipid A. Representative negative ion MALDI-TOF lipid A mass spectra of the different samples analyzed. Right panel: proposed structures of the main molecular species present obtained. All strains tested, both in the absence or in the presence of sub-lethal [PxB], displayed identical lipid A structure. (B) Growth of NTHi1a, 1b, 1c and Rd KW20 in rich medium. Cells were grown in sBHI broth in a BioScreen incubator with the OD600 nm recorded every 10 min. Each line shows the mean of 8 to 12 replicates for one strain. Optical densities lower than 0.01 are not shown. To account for differences in inoculum sizes, the lines have been shifted along the x axis to superimpose the growth curves of all strains. Color code: NTHi1a, red; 1b, yellow; 1c, green; Rd KW20, blue. (C) Transformation frequencies of H. influenzae strains NTHi1a, 1b and 1c. Vertical bars represent the means of at least three biological replicates ± standard deviations. We did not observe significant differences. (D) Bacterial loads in the lungs of CD1 mice infected by NTHi1a, 1b and 1c. Mice were infected intranasally with 108 bacteria. Bacterial counts in lungs at 24 or 48 h PI were determined. Results are reported as log10 CFU/lung. Lines inside boxes represent median values. Statistical differences could not be observed among isolates NTHi1a, 1b and 1c. In the three cases, bacterial recovery at 48 (white box) was significantly lower than at 24 (grey box) h PI (p (PDF)