Effect of Porphyromonas gingivalis PrtC on cytokine expression in ECV304 endothelial cells and its level in subgingival plaques from patients with chronic periodontitis

Abstract

Aim: To investigate the effect of the collagenase gene (prtC) product of Porphyromonas gingivalis on inducing host cells to secrete inflammatory cytokines, and to discuss the correlation between the PrtC level in subgingival plaque samples and clinical parameters. Methods: A prokaryotic expression system pET32a-prtC-Escheria coli BL21DE3 was constructed. Antigenicity and immunoreactivity of the recombinant PrtC protein (rPrtC) was identified by Western blotting. ELISA was applied to detect interleukin (IL)-1α, IL-8, and TNF-α levels in supernatants from rPrtC-induced human umbilical vein endothelial cells (HUVEC) originated ECV304 cells . Clinical parameters recorded at baseline and after treatment included bleeding on probing (BOP), probing depth (PD), and attachment loss (AL). ELISA was established to measure the PrtC level in 196 subgingival plaque samples from 49 patients with chronic periodontitis. Results: After coincubation with 1 µg/mL rPrtC for 24 h and with 5 or 10 µg/mL rPrtC for 12 h, the levels of IL-1α, IL-8, and TNF-α secreted by the ECV304 cells increased significantly (P<0.05). The PrtC level in the BOP-positive or the ≥5 mm AL or >6 mm PD sites was higher than that in the BOP-negative or the ≤2 mm AL or ≤6 mm PD sites (P<0.05), respectively. Compared with baseline, the PrtC levels in different AL sites or in the ≤6 mm PD pockets decreased remarkably after treatment (P<0.01), but in the BOP-positive or in the >6 mm PD sites, the PrtC levels changed insignificantly (P>0.05). Conclusion: rPrtC is able to directly induce host cells to synthesize and secrete IL-1α, IL-8, and TNF-α. The PrtC level in subgingival samples is correlated with BOP, AL, and PD.

Genome Sequence of Poultry Pathogen Riemerella anatipestifer Strain RA-YM

“…The product of hagA1, a gene encoding a serine protease/hemagglutinin identified in the RA-YM genome, contains a 280-amino acid C-terminal region with 43% similarity to the HagA of Porphyromonas gingivalis, which may play a pivotal role in host colonization by R. anatipestifer and, thus, in infection (7). The gene prtC, which encodes an extracellular collagenase that could induce host cells to synthesize and secrete inflammatory cytokines to exert a destructive effect on periodontal tissue by P. gingivalis, may play an important role in RA-YM's pathogenicity (17). Five gene clusters involved in polysaccharide biosynthesis and export were identified, and the products of a gene cluster which are similar to alginate O-acetyltransferases of P. aeruginosa probably function in biofilm formation (6).…”

Protease activities of vaginalPorphyromonasspecies disrupt coagulation and extracellular matrix in the cervicovaginal niche

“…As such, these enzymes are unlikely to be found in culture supernatants. The collagenase activity of the P. gingivalis U32 collagenase, PrtC, has been confirmed (84,85,128), but these studies used cell lysates (85), recombinant PrtC (84) or heterologous expression systems (128). To our knowledge, no studies have determined the subcellular localization of PrtC, and sequence analysis does not reveal any indication of secretion signals.…”

Molecular Characterization, Virulence and Antimicrobial Susceptibility Testing of <i>Riemerella anatipestifer</i> Isolated from Ducklings

“…isolate of R. anatipestifer serotype 1) (Wu et al 2007) . Outer membrane protein A (OmpA) is another virulence factor play a very important role in virulence and has been reported to be immunogenic proteins (Hu et al 2012;Huang et al 2002) .…”