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Coating and corruption of human neutrophils by bacterial outer membrane vesicles

ABSTRACT: Porphyromonas gingivalis is a keystone oral pathogen that successfully manipulates the human innate immune defenses, resulting in a chronic pro-inflammatory state of periodontal tissues and beyond. Here we demonstrate that secreted outer membrane vesicles (OMVs) are deployed by P. gingivalis to selectively coat and activate human neutrophils, thereby provoking degranulation without neutrophil killing. Secreted granule components with antibacterial activity, especially LL-37 and MPO, are subsequently degraded by potent OMV-bound proteases known as gingipains, thereby ensuring bacterial survival. In contrast to neutrophils, the P. gingivalis OMVs are efficiently internalized by macrophages and epithelial cells. Importantly, we show that neutrophil coating is a conserved feature displayed by OMVs of at least one other oral pathogen, namely Aggregatibacter actinomycetemcomitans. Altogether, we conclude that P. gingivalis deploys its OMVs for a neutrophil-deceptive strategy to escape phagocytosis and to create a favorable inflammatory niche.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (human) Porphyromonas Gingivalis (strain Atcc Baa-308 / W83)

SUBMITTER: Claudia Hirschfeld

LAB HEAD: Prof. Dr. Dörte Becher

PROVIDER: PXD026704 | Pride | 2022-08-16

REPOSITORIES: pride

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			Action	DRS
	200925_QE1_PM1_CHi_JM_MDT_MPO_01.raw	Raw
	200925_QE1_PM1_CHi_JM_MDT_MPO_02.raw	Raw
	200925_QE1_PM1_CHi_JM_MDT_MPO_03.raw	Raw
	200926_QE1_PM1_CHi_JM_MDT_PPAD_01.raw	Raw
	200926_QE1_PM1_CHi_JM_MDT_PPAD_02.raw	Raw

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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.

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