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Oral Pathogen ‘Neutralizes’ the Human Innate Immune Defence

ABSTRACT: Our present findings show for the first time that the virulence factor PPAD protects the oral pathogen P. gingivalis against neutrophil insults at three distinct levels, namely phagocytosis, NETosis and CAMP activity. This identifies PPAD as a major agent in the evasion of human innate immunity, a view that is supported by studies from Potempa and co-workers showing PPAD-dependent citrullination of complement29. Why is this important? Every day, millions of people lose teeth due to periodontitis. This has a serious impact on their quality of life, not only for aesthetic reasons but much more so because tooth loss causes difficulties in eating and interferes with interpersonal communication. Yet, only few people realize that there is a major bacteriological component in periodontitis where our immune defences are breached with potentially serious implications, especially systemic and chronic diseases ranging from bacteraemia to rheumatoid arthritis.

INSTRUMENT(S): LTQ Orbitrap Elite

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Culture, Macrophage

SUBMITTER: Thomas Sura

LAB HEAD: Doerte Becher

PROVIDER: PXD010798 | Pride | 2018-10-15

REPOSITORIES: Pride

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			Action	DRS
	160614_VE1_PM4_ThS_TSt_Neutrophile_Kontrolle_A_180_min.raw	Raw
	160615_VE1_PM4_ThS_TSt_Neutrophile_Kontrolle_B.raw	Raw
	160615_VE1_PM4_ThS_TSt_Neutrophile_Kontrolle_C.raw	Raw
	160615_VE1_PM4_ThS_TSt_Neutrophile_W83_A.raw	Raw
	160615_VE1_PM4_ThS_TSt_Neutrophile_W83_B.raw	Raw

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Oral Pathogen ‘Neutralizes’ the Human Innate Immune Defence

Project description:Our present findings show for the first time that the virulence factor PPAD protects the oral pathogen P. gingivalis against neutrophil insults at three distinct levels, namely phagocytosis, NETosis and CAMP activity. This identifies PPAD as a major agent in the evasion of human innate immunity, a view that is supported by studies from Potempa and co-workers showing PPAD-dependent citrullination of complement.

2018-10-15 | PXD009081 | Pride

The Extracellular Proteome and Citrullinome of the Oral Pathogen Porphyromonas gingivalis

Project description:Porphyrmonas gingivalis is an oral pathogen associated with the inflammatory disease periodontitis. The colonization of oral epithelial surfaces by P. gingivalis may also lead to the autoimmune disease rheumatoid arthritis. One of the hallmarks of rheumatoid arthritis is the loss of tolerance against citrullinated proteins. Citrullination is a post-translational modification of arginine residues, leading to a change in structure and function of the respective protein. This modification is catalysed by peptidylarginine deiminases (PAD). Interestingly, P. gingivalis secretes a citrullinating enzyme, known as P. gingivalis PAD (PPAD), which targets bacterial and human proteins. Since the extent of P. gingivalis protein citrullination by PPAD was not yet known, the present study was aimed at identifying the extracellular proteome and citrullinome of P. gingivalis. To this end, extracellular proteins of two reference strains, two PPAD-deficient mutants and three clinical isolates of P. gingivalis were analysed by mass spectrometry. The results uncovered substantial heterogeneity in the extracellular proteome and citrullinome of P. gingivalis, especially in relation to the extracellular detection of typical cytoplasmic proteins. In contrast, major virulence factors were identified in all investigated isolates although their citrullination was shown to vary. This may be related to post-translational processing of the PPAD enzyme.

2018-02-28 | PXD003444 | Pride

Transcriptome profiling of gingival fibroblasts treated by vitamin D3

Project description:In humans, vitamin D3 is a secosteroid, a prohormonal precursor of biologically active hydroxyforms. Vitamin D3 is related to periodontitis. The third American Health and Nutrition Survey found that plasma 25(OH)D3 levels are negatively correlated with attachment loss in people aged >50 years. The relationship between vitamin D3 and periodontitis remains to be investigated. We sent human gingival fibroblasts treated by vitamin D3 for transcriptome sequencing.

2022-09-09 | GSE212728 | GEO

Thioredoxin-1 promotes the restoration of alveolar bone in periodontitis with diabetes

Project description:Treatment of periodontitis in people with diabetes remains challenging. Diabetes-enhanced oxidative stress is a primary cause of aggravation of periodontitis. The present study aimed to investigate the therapeutic potential of thioredoxin-1 (TRX1), an essential endogenous antioxidant protein, in the management of periodontitis with diabetes, as well as its role in modulating osteogenic differentiation. Our findings indicated that the production of reactive oxygen species (ROS) was elevated, while the expression of TRX1 was significantly reduced in the periodontal tissues of periodontitis mice with diabetes. Furthermore, knockdown of TRX1 in periodontal ligament stem cells (PDLSCs) resulted in the inhibition of osteogenic differentiation through disrupting Wnt/β-catenin signal pathway. However, this inhibition was restored upon administration of recombinant human TRX1 (rhTRX1). Importantly, rhTRX1 treatment decreased ROS generation, activated Wnt/β-catenin signal pathway and considerably promoted the alveolar bone repair of periodontitis mice with diabetes. These findings highlighted the crucial protective role of TRX1 in periodontitis with diabetes and suggested that it may serve as a potential therapeutic target for refractory periodontitis associated with oxidative stress.

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Single-Cell and Spatially Resolved Interactomics of Tooth-Associated Keratinocytes in Periodontitis

Project description:Periodontitis affects billions of people worldwide. To address relationships of periodontal niche cell types and microbes in periodontitis, we generated an integrated single-cell RNA sequencing (scRNAseq) atlas of human periodontium (34-sample, 105918-cell), including sulcular and junctional keratinocytes (SK/JKs). SK/JKs displayed altered differentiation states and were enriched for effector cytokines in periodontitis. Single-cell, metagenomics revealed 37 bacterial species with cell-specific tropism. Fluorescence in situ hybridization detected intracellular 16S and mRNA signals of multiple species and correlated with SK/JK proinflammatory phenotypes in situ. Cell-cell communication analysis predicted keratinocyte-specific innate and adaptive immune interactions. Highly multiplexed immunofluorescence (33-antibody) revealed peri-epithelial immune foci, with innate cells often spatially constrained around JKs. Spatial phenotyping revealed immunosuppressed JK-microniches and SK-localized tertiary lymphoid structures in periodontitis. Here, we demonstrate impacts on and predicted interactomics of SK and JK cells in health and periodontitis, which requires further investigation to support precision periodontal interventions in states of chronic inflammation.

2024-06-05 | GSE266897 | GEO

Differential virulence of Aggregatibacter actinomycetemcomitans serotypes explained by exoproteome heterogeneity

Project description:Aggregatibacter actinomycetemcomitans (Aa) is a Gram-negative bacterial pathogen associated with periodontitis and non-oral diseases like rheumatoid arthritis and Alzheimer´s disease. Aa isolates with the serotypes a, b and c are globally most prevalent. Importantly, isolates displaying these serotypes have different clinical presentations. While serotype b isolates are predominant in severe periodontitis, serotypes a and c are generally encountered in mild periodontitis or healthy individuals. It was so far not known how these differences are reflected in the overall secretion of virulence factors. Therefore, this study was aimed at a comparative analysis of exoproteomes from different clinical Aa isolates with serotypes a, b or c by mass spectrometry, and a subsequent correlation of the recorded exoproteome profiles with virulence. Overall, we identified 425 extracellular proteins. Significant differences in the exoproteome composition of isolates with different serotypes were observed in terms of protein identification and abundance. In particular, serotype a isolates presented more extracellular proteins than serotype b or c isolates. These differences are mirrored in their virulence in infection models based on human salivary gland epithelial cells and neutrophils. Remarkably, serotype a isolates displayed stronger adhesive capabilities and induced more lysis of epithelial cells and neutrophils than serotype b or c isolates. Conversely, serotype c isolates showed relatively low leukotoxicity, while provoking NETosis to similar extents as serotype a and b isolates. Altogether, we conclude that the differential virulence presentation by Aa isolates with the dominant serotypes a, b or c can be explained by their exoproteome heterogeneity.

2022-12-16 | PXD036160 | Pride

Using SWATH-MS to Identify New Molecular Biomarkers in Gingival Crevicular Fluid for Detecting Periodontitis and its Response to Treatment

Project description:Over the last years, proteomic techniques have been demonstrated to be valuable tools for analysing the periodontal proteome in gingival crevicular fluid (GCF) samples contributing to identifying extensive and comprehensive protein profiles associated with periodontitis. However, the periodontal proteome has not yet been studied using sequential window acquisition of all theoretical mass spectra (SWATH-MS). Consequently, the objective of the present longitudinal research was to investigate the periodontal proteome in GCF in three clinical conditions (periodontal health, untreated periodontitis, and treated periodontitis), applying SWATH-MS to 1) analyse the periodontal proteome structure in the three clinical groups, 2) compare the differential protein expression between the three clinical groups, and 3) evaluate the diagnostic accuracy of the proteins quantified to identify new biomarkers with a high capacity to discriminate the three clinical conditions. A total of 84 subjects, 44 periodontally healthy and 40 with generalised untreated periodontitis in stages III-IV, were included, and GCF samples were collected. The diagnosis of periodontal status was conducted by two experienced dentists using clinical and radiographic criteria. The definition of periodontal status was established following the Classification of Periodontal Diseases and Conditions published in 2018. In the untreated periodontitis group, 38 underwent conventional non-surgical treatment and were evaluated clinically after two months (treated periodontitis group). In the revaluation, 25 individuals showed significant clinical improvement in probing pocket depth (PPD) at the selected sites, and GCF samples were collected from them.

2024-10-17 | PXD043474 | Pride

Human Gingival Crevicular Fluid Reveals the Occurrence of Ferroptosis During Clinical Progression of Periodontitis

Project description:Thirteen healthy men diagnosed with periodontitis were enrolled. Progression of periodontitis was monitored by clinical attachment level (CAL) changes at the site level over twelve weeks via weekly clinical evaluations and gingival crevicular fluid (GCF) samples were collected using cellulose strips for subsequent mass spectrometry analysis. Progression group (PG) included 18 GCF samples matching sites with disease progression (CAL ≥ 2 mm), while the non-Progression group (NP) included 18 GCF samples from stable sites (No CAL changes) matched in the same patients. Clinical metrics, HPRLC-MS/MS, in silico methods, and bioinformatics tools, were employed to analyze the proteome of GCF, revealing the significant occurrence of ferroptosis during clinical progression of periodontitis.

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