maltophilia strains isolated from CF patients were shown Tozasertib in vitro to be able, although with striking differences, to adhere to and form learn more biofilm on polystyrene [20]. Since information on the ability of S. maltophilia to grow as biofilm in CF airway tissues is scarce, in the study described in this paper we evaluated, by quantitative assays and microscopic analysis (scanning electron and confocal laser microscopy), the ability of CF S. maltophilia strains to adhere, invade and form biofilm on CF-derived IB3-1 bronchial epithelial cell monolayers. Moreover, the role of flagella in adhesiveness on IB3-1 epithelial cells was also evaluated
by the construction of two independent S. maltophiia fliI deletion mutants that were used to infect cultured monolayers. Some of the results of the present study have been previously presented in the form of an abstract at the 18th European Congress of Clinical Microbiology and Infectious Diseases [21]. Results S. maltophilia is able to adhere to and form biofilm on IB3-1 cell monolayers We used IB3-1 human bronchial CF-derived cells to investigate the ability of S. maltophilia to adhere to and form biofilm. Confluent IB3-1 cell monolayers were independently infected with the 12 CF-derived S. maltophilia strains chosen for this study (Table 1); both the adhesiveness and the ability to form biofilm were measured by determining the number (cfu) of bacteria 2 and
24 hours post-infection, respectively. Growth curves, obtained with bacteria grown in AZD1480 trial MH broth, showed no significant differences in the mean generation time between isolates (mean ± SD: 3.35 ± 0.39 hours). Table 1 Microbiological features of S. maltophilia OBGTC strains (n = 12) used in this study. Strain Patient agea Co-isolated with: Chronic lung infection isolateb Past P. aeruginosa infection OBGTC5 oxyclozanide 13 Pa, Ca – + OBGTC9 17 Sa + + OBGTC10 13 only + – OBGTC20 11 Pa + + OBGTC26 11 only – - OBGTC31 16 Pa, Sa + + OBGTC37 3 only – NA OBGTC38 9 Sa – + OBGTC44 16 Pa + + OBGTC49 5 NA + + OBGTC50 10 NA + + OBGTC52 25 only + + Caption and Abbreviations:aAges shown are in years at the time of strain isolation.
b Chronic infection is defined as the presence of two or more positive cultures for S. maltophilia in a year. Pa: P. aeruginosa; Ca: C. albicans; Sa: S. aureus; NA, not available. All S. maltophilia strains tested were able to adhere to IB3-1 cells after 2 hours of incubation, with significantly different levels of adhesiveness among the strains (Figure 1A). S. maltophilia strains OBGTC9 and OBGTC10 showed the highest levels of adhesiveness (5.6 ± 1.2 × 106 and 5.0 ± 1.1 × 106 cfu chamber-1, respectively; P > 0.05), significantly higher if compared to that of the other strains (P < 0.001). Figure 1 Adhesion to and biofilm formation on IB3-1 cell monolayer of clinical isolates of S. maltophilia from CF patients. A. Adhesion levels of S. maltophilia to IB3-1 cell monolayers.
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