All other CoNS (n = 25) were ica – biofilm- 20-kDaPS- All ica +

All other CoNS (n = 25) were ica – biofilm- 20-kDaPS-. All ica + biofilm+ S. epidermidis strains were PIA-positive by specific immunofluorescence test, whereas, ica – biofilm- or ica + biofilm – strains Cyclosporin A in vivo were PIA-negative. In our S. epidermidis strain collection, 46% (n = 23) were PIA positive and 60% (n = 30) were 20-kDaPS positive. IcaADBC prevalence in our collection was 68%, whereas 46% of S. epidermidis strains were biofilm-producing. 20-kDaPS expression among ica + S. epidermidis strains was 70% (24 ica + 20-kDaPS+

amongst 34 ica + S. epidermidis strains), whereas, 20-kDaPS expression among ica – strains was 37% (6 ica – 20-kDaPS + amongst 16 ica – S. epidermidis strains). 20-kDaPS expression in relation to biofilm formation reveals that 78% of biofilm-producing S. epidermidis strains expressed 20-kDaPS (18 biofilm + 20-kDaPS + in 23 biofilm + S. epidermidis strains), whereas, 44% of biofilm-negative strains were 20-kDaPS positive (12 biofilm- 20-kDaPS+ of 27 biofilm- S. epidermidis strains). These results show

that the majority of clinical S. epidermidis isolates express 20-kDaPS and that there is no strict correlation of icaADBC-genotype or biofilm phenotype and expression of 20-kDaPS. Expression of 20-kDaPS and PIA by S. epidermidis strains with known genetic backgrounds Using an indirect immunofluorescence test with specific anti-PIA Farnesyltransferase antiserum S. epidermidis strains 1457, 8400, and 9142 were shown to express PIA, while the isogenic icaA-insertion mutants 1457-M10, find more M24 and 8400-M10 and isogenic icaC-insertion mutants M22 and M23 did not express PIA. Similarly, S. epidermidis 5179, 5179R1 and 1585 did not synthesize PIA as in

the former two strains icaADBC is inactivated through insertion of IS257[37], while 1585 is icaADBC-negative. Using specific anti-20-kDaPS antiserum S. epidermidis 1457, 1457-M10, M22, M23, M24, 8400, 8400-M10, 9142, 5179, 5179R1 were 20-kDaPS positive, whereas, S. epidermidis strain 1585 was 20-kDaPS negative. A representative immunofluorescence test with anti-PIA and anti-20-kDaPS antisera, Compound C supplier comparing S. epidermidis 1457 and 1457-M10, is displayed in Figure 1. An identical expression pattern of 20-kDaPS was independently demonstrated for these strains using specific ELISA, excluding that there are significant quantitative differences in 20-kDaPS antigen expression between the isogenic mutant strain pairs (Figure 2). 20-kDaPS detection in transposon mutants of S. epidermidis 1457-M10, M22, M23, M24 is shown in Figure 3. Inactivation of icaA in mutant 1457-M10 and of icaC in mutants M22 and M23 lead to biofilm negative and PIA negative phenotype, but did not alter 20-kDaPS antigen detection.

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