To examine the role of CPS, both the wild-type

and the epsC mutant were used in an in vitro challenge of primary human gingival fibroblasts. Since the epsC mutant has altered physical properties, it was important to compare the sedimentation rate and viability of both the wild type and the mutant strain since these could have influenced the amount of living bacterial cells that are in contact with the fibroblasts. p38 MAPK inhibitor No differences were observed between the strains during the 6 hours of infection. From the infection experiments of the gingival fibroblasts it became apparent that pro-inflammatory mediators IL-1β, IL-6 and IL-8 expression levels were up-regulated after a 6-hour challenge with both wild-type W83 and the epsC mutant in comparison to the non-infected control, especially when MOIs of 10.000:1 were used. A challenge with the epsC mutant induced a significantly higher pro-inflammatory immune response than

a challenge with the wild type W83, as shown by IL-1β, IL-6 and IL-8 gene expression. So, even though purified P. gingivalis CPS has been shown to stimulate pro-inflammatory cytokine expression in murine peritoneal macrophages [11] the absence of capsule induces extra cytokine induction when viable P. gingivalis cells selleck were used to challenge fibroblasts. Capsular polysaccharides of several bacteria have been implicated in down-regulation of pro-inflammatory cytokine production, including Klebsiella pneumonia [29]. Bacteroides fragilis capsular polysaccharide complex has been shown to induce IL-10 expression, a regulating cytokine which may cause suppression of the immune system [30]. An explanation of our results may be that the Farnesyltransferase CPS prevents more potent immune inducers to be recognized by Toll-like MI-503 in vivo receptors on the fibroblasts.

It has been shown that the capsular antigen in Salmonella typhi, referred to as Vi-antigen, is able to prevent Toll-like receptor 4 recognition of LPS, thereby reducing expression of pro-inflammatory TNF-α and IL-6 [31–33]. In E. coli the capsule may cover short (10 nm) bacterial adhesins, which do not penetrate the 0.2-1.0 μm capsular layer, preventing them from being recognized by the immune system [26]. Likewise, P. gingivalis strain W83 was described as to have a small amount of short fimbriae that might be mostly covered by the CPS [34]. Another or additional explanation of our findings could be immune suppression by P. gingivalis CPS, meaning that CPS would actively modulate the immune response of the fibroblasts, leading to lower inflammatory cytokine expression levels, potentially enabling P. gingivalis to evade the immune system. For several bacteria it has been described that capsular biosynthesis can be modulated depending on environmental conditions [35, 36]. Although presently no regulation of P. gingivalis capsule expression has been described, we can not exclude the possibility that in the in vivo situation capsule expression is regulated.