Research progress on the pathogenicity of Porphyromonas gingivalis clinical strains / 口腔疾病防治

@#Porphyromonas gingivalis (P. gingivalis) is closely related to the occurrence and development of periodontitis. It is considered to be one of the important pathogens leading to alveolar bone resorption. At present, research on P. gingivalis mostly adopts standard laboratory strains whose genetic characteristics have been confirmed, are guaranteed and are traceable, such as ATCC 33277. The virulence phenotypes (endotoxin, firmbria, etc.) of clinically extracted isolates are quite different from those of standard strains, and the pathogenic effects and ability of the host are also widely different. In addition, P. gingivalis is considered to have a significant correlation with a variety of systemic diseases, and the virulence characteristics and pathogenic ability of different strains will have different effects on systemic diseases. However, at present, there is a lack of research on clinical strains and standard strains, and there is a lack of systematic comparison between the two sources of bacteria. In this paper, the differences in the virulence phenotypes and pathogenic effects between clinical isolates and standard strains of P. gingivalis in the last 5-10 years are reviewed. The aim is to elucidate the important virulence gene loci in the P. gingivalis gene sequence, which will play an important role in improving therapeutic methods and the development of related drugs.

The virulence system of Porphyromonas gingivalis: Genes, mechanism and potential role of gingipains inhibitors

ABSTRACT@#The oral microbiome comprises several hundreds of bacterial species that contribute to periodontitis, the most complex polymicrobial inflammatory disorder. Porphyromonas gingivalis is a prominent periodontitis pathogen that produces gingipains as a major virulent factor. Gingipain facilitates P. gingivalis survival, pathogenicity, and growth. Several genes were identified to have a role in the regulating of P. gingivalis pathogenesis. Studies suggest that gingipains inhibition is key for the successful treatment of periodontitis. As of now, several gingipain inhibitors have been developed, some exhibit high inhibition activity against gingipains. However, most inhibitors offer unknown toxicity and undesirable side effects. Hence, the development of highly potent and safe gingipain inhibitor is a major concern for periodontitis treatment. The present review highlights the connectivity between P. gingivalis, virulent factors, and its gene, periodontitis, and gingipain inhibitors. Development of gingipains inhibitors would not only treat periodontitis but would also assist in the treatment of other associated systemic diseases, for example: rheumatoid arthritis, cardiovascular diseases, diabetes, and Alzheimer's disease.

Ca mobilization and signaling pathways induced by rRgpB in human gingival fibroblast / 浙江大学学报·医学版

: To assess the () recombinant gingivalis gingipain R2 (rRgpB)-induced Ca mobilization in human gingival fibroblast (HGF) mediated by protease-activated receptor (PAR) and its downstream signal transduction pathways. : Flow cytometry was used to detect the expression of PAR in HGF. The proliferation of HGF was measured by CCK-8. The dynamic changes of intracellular Ca concentration in HGF induced by rRgpB and the blocking effect of PAR-1 antagonist were observed by laser confocal microscopy. Western blot was performed to determine the phosphorylation levels of c-Jun N-terminal kinase (JNK), extracellular signal-regulated kinase (ERK) 1/2, p38 mitogen-activated protein kinase (p38 MAPK) and p65 in HGF. : PAR-1 and PAR-3 were expressed in HGF, and the rRgpB could promote the proliferation of HGF. rRgpB caused a transient increase in [Ca], which could be completely suppressed by vorapaxar, a PAR-1 antagonist. The phosphorylation levels of JNK, ERK1/2 and p65 were significantly up-regulated after the induction of rRgpB for and (all <0.05), which was completely inhibited by vorapaxar. However, the phosphorylation level of p38 MAPK had no significant change after rRgpB stimulation. : rRgpB causes an increase in [Ca] in HGF mediated by PAR-1. JNK, ERK1/2 and nuclear factor-κB may be involved in intracellular signal transduction after PAR-1 activation.

Experimental research on Arginine-gingipain A gene vaccine from Porphyromonas gingivalis that prevents peri-implantitis in Beagle dogs / 华西口腔医学杂志

OBJECTIVE@#This study aims to use Arginine-gingipain A gene vaccine (pVAX1-rgpA) to immunize adult Beagle dogs and to evaluate its effect during peri-implantitis progression and development.@*METHODS@#Plasmid pVAX1-rgpA was constructed. The second and third bilateral mandible premolars of 15 adult Beagle dogs were extracted, and the implants were placed immediately. After 3 months, the animals were randomly divided into groups A, B, and C. Afterward, the animals were immunized thrice with plasmid pVAX1-rgpA, with heat-killed Porphyromonas gingivalis, or pVAX1, respectively. IgG in the serum and secretory IgA (sIgA) in saliva were quantitatively analyzed by enzyme-linked immunosorbent assay before and after 2 weeks of immunization. Peri-implantitis was induced with cotton ligatures fixed around the neck of implants. Probing depth (PD) and bleeding on probing were recorded. All animals were sacrificed after ligaturation for 6 weeks. Decalcified sections with thickness of 50 μm were prepared and dyed with methylene blue to observe the bone phenotype around implants.@*RESULTS@#Levels of serum IgG and sIgA in saliva were higher in groups A and B after immunization than before the process (P0.05). At 4 and 6 weeks after ligaturation, PD of the ligatured side in group C was higher than that in groups A and B (P0.05). Bone loss in group A was significantly lower than that of the other groups (P<0.05). Abundant inflammatory cells and bacteria were present in the bone loss area around the implants in the three groups, as identified through hard tissue section observation. However, group C presented the most number of inflammatory cells and bacteria in the bone loss area around the implants.@*CONCLUSIONS@#IgG and sIgA can be generated by immunity with rgpA DNA vaccine, which can significantly slow down bone loss during experimental peri-implantitis in dogs.

Research progress on the type Ⅸ secretion system of Porphyromonas gingivalis / 华西口腔医学杂志

In recent years, the study found that Porphyromonas gingivalis type Ⅸ secretion system (T9SS) is a novel protein secretion system, also known as Por secretion system (PorSS). Unlike the eight protein secretion systems found in the past, the system is a polyprotein complex found only in Bacteroides. The secreted proteins have both N- and C-terminus, where the former includes Sec-dependent type Ⅰ signals peptide, and the latter contains conserved domains (C-terminal conserved domain, CTD). Porphyromonas gingivalis T9SS includes proteins such as intima, outer membrane, cytoplasm, and cell cycle, including at least 34 proteins containing CTD. Porphyromonas gingivalis T9SS is involved in regulating associated virulence factors including gingivin, fimbriae, lipopolysaccharide, HBP35, CPG70 protein and peptidyl-arginine deiminase. These CTD-containing virulence proteins are localized by T9SS and then released to the extracellular domain, thereby destroying periodontal tissue. Therefore, this review summarizes the research progress on the T9SS of Porphyromonas gingivalis.

Genotipificación de los genes rgpA y kgp que codifican para las gingipaínas de Porphyromonas gingivalis / Genotyping of rgpA and kgp genes encoding Pophyromonas gingivalis gingipains

Porphyromonas gingivalis es un microorganismo fuertemente asociado con la etiología de la periodontitis. Esta bacteria posee varios factores de virulencia, dentro de los que destacan las gingipaínas, debido a sus múltiples acciones relacionadas con la destrucción de la matriz extracelular del tejido conectivo periodontal, la modulación del sistema inmune del hospedero y la estimulación de la expresión de citoquinas pro-inflamatorias. Estas proteinasas tienen afinidades específicas siendo Arg-gingipaínas (RgpA y RgpB, codificadas por los genes rgpA y rgpB, respectivamente) y Lys-gingipaínas (Kgp, codificada por el gen kgp). Se ha descrito que existen polimorfismos en los genes que codifican para esta proteinasas. El objetivo del presente estudio fue describir la frecuencia de los genotipos identificados para los genes rgpA y kgp en aislados clínicos de P. gingivalis, obtenidos desde pacientes con periodontitis. Para ello se utilizó amplificación por PCR de los genes rgpA y kgp, seguido de análisis de restricción. De un total de 47 aislados provenientes de 4 individuos con periodontitis crónica y 2 con periodontitis agresiva, se genotipificaron 38 aislados para el gen rgpA, exhibiendo la totalidad de éstos el patrón electroforético A (100 por ciento). Para el gen kgp se genotipificaron 43 aislados, presentando 28 de ellos (65.2 por ciento) el perfil electroforético kgp-I y 15 aislados (34.8 por ciento) el perfil kgp-II. En los aislados provenientes de un individuo fue posible apreciar ambos genotipos descritos para el gen kgp. Los resultados indican un predominio del patrón electroforético A (rgpA) y que el genotipo kgp-I fue el más frecuentemente encontrado de los genotipos kgp.

Porphyromonas gingivalis is a microorganism strongly associated with the etiology of periodontitis. This periodontal bacterium possesses an array of virulence factors, among which gingipains have a key importance, being involved with extracellular matrix destruction of periodontal tissues, modulation of host immune response and stimulation in the production of pro-inflammatory cytokines by different types of cells. These proteinases have specific affinities, being Arg-gingipains (RgpA and RgpB, encoded by rgpA and rgpB genes, respectively) and Lys-gingipains (Kgp, encoded by the kgp gene). It has been described that there are polymorphisms in the genes encoding for gingipains. Therefore, the aim of the present study was to describe the frequency of rgpA and kgp genotypes in clinical isolates of P. gingivalis obtained from periodontitis patients. For determining the rgpA and kgp genotypes, we used PCR amplification and restriction analysis. From 47 isolates obtained from 4 individuals with chronic periodontitis and 2 subjects with aggressive periodontitis, 38 were typified for rgpA gene and all exhibited the electrophoretic pattern A (100 percent). For kgp gene, we characterized 43 isolates, 28 of them (65.2 percent) with the kgp-I electrophoretic profile and 15 isolates (34.8 percent) with the kgp-II profile. In the isolates belonging to one individual, we found both genotypes of kgp gene. The results indicate a clear predominance of the electrophoretic pattern A (for rgpA gene) and kgp-I genotype was the most frequently found of the kgp genotypes.

Expression of gingipain K in Porphyromonas gingivalis in the subgingival plaque of puberty gingivitis / 实用口腔医学杂志

Objective:To detect and compare the intensity of gingipain K(Kgp)in culture medium and cell extract of Porphyromonas gingivalis(P.gingivalis)isolates in puberty gingivitis,and then to reveal the possible relationship between Kgp and puberty gingivitis.Methods:36 patients with puberty gingivitis aged from 14 to 17 years were enrolled.Clinical parameters including GI,SBI and PD were evaluated before subgingival plaque samples collection.Subgingival plaque samples were collected and then P.gingivalis isolates were obtained.16S rRNA PCR was used to confirm the presence of P.gingivalis in clinical isolates.Bacteria were cultured in BHI agar base and harvested at the end of log-phase growth.Culture fractions of P.gingivalis(culture medium and cell extracts)were performed with SDS-PAGE and Western blot technique using primary antibody against specific anti-Kgp N-terminal IgG subdomain.The data were statistically analyzed using SPSS 11.5 software.The relationship between the Kgp intensity and the clinical parameters was statistically analyzed using sum rank test.Results:There was positive correlation between the intensity of Kgp N-terminal IgG subdomain and the clinical parameters(P