These findings were consistent with the primary microarray-based data of this study and expression data of S. aureus Mu50 [10]. Figure 2 Transcript quantification of the essential capsule biosynthesis gene cap5E by real

time PCR. a) Transcript amounts of cap5E throughout the growth curve of hVISA SA137/93A (filled square), VISA SA137/93G (filled triangle), VSSA Newman (filled circle) and VSSA SA1450/94 (filled diamond) indicated as copy number per 106 copies of the housekeeping gene gyrB. b) Transcript amounts of cap5E of VSSA strains (R: Reynolds*, N: Newman, 14: SA1450/94) and VISA strains (A: SA137/93A*, G: SA137/93G*, Mu: Mu50) at OD600 = 1 and OD600 = 4–5 indicated as copy number per 106 copies of gyrB. * Error bars are not visible at OD600 = 1 because of minimal data variations.

The CPs of Klebsiella Crenolanib molecular weight pneumoniae were found to PF-02341066 purchase contribute to resistance to cationic defensins, lactoferrin, protamine sulfate and polymyxin B, and in this context, the capsule was assumed to protect bacteria by limiting the interaction of the antimicrobial peptides with the surface [54]. Later, similar results were obtained with polysaccharides from Streptococcus pneumoniae Selleckchem BAY 73-4506 and alginate from Pseudomonas aeruginosa [55]. A possible role of CPs in vancomycin resistance has repeatedly been discussed in the literature. Boyle-Vavra et al. found that susceptible passage revertants of the CP5 producing VISA isolates MI, NJ and PC were no longer CP typable, while passaging in presence of vancomycin retained the CP phenotype [56]. Besides, comparative expression profiling experiments on VISA isolates Mu50, MI, JH9 and their respective

susceptible parent or mutant strains showed that some (but not necessarily all) of the genes of the type 5 capsule were more highly FAD expressed in the VISA strains [10, 45]. Enhanced capsule production in other VISA was also reported [57] and deletion of the yabJ-spoVG operon affected glycopeptide susceptibility and capsule production in S. aureus simultaneously [50]. Taken together, these findings encouraged us to further investigate the role of CPs in vancomycin resistance. Detection of the capsule by immunofluorescence Production of CP5 was analysed by immunofluorescent labelling of cells of SA137/93G and the susceptible strains SA1450/94 and Newman after 6 h of incubation in LB. The results revealed that the VISA strain produced higher amounts of CP5 than SA1450/94 and S. aureus Newman (Figure 3). Figure 3 Comparison of CP5 production in a VISA and two VSSA strains. CP5 was labelled by immunofluorescence (CY3, green). As a control, all cells were stained using DAPI (blue). Cells were grown for 6 h in LB at 37°C. a) VISA SA137/93G; b) control strain SA1450/94; c) S. aureus Newman.