Vanessa Tubero Euzebio Alves, Tomaz Alves, Emanuel Silva Rovai, Hatice Hasturk, Thomas Van Dyke, Marinella Holzhausen, Alpdogan Kantarci
{"title":"精氨酸特异性鞘脂素（RgpA/RgpB）敲除可调节中性粒细胞机制。","authors":"Vanessa Tubero Euzebio Alves, Tomaz Alves, Emanuel Silva Rovai, Hatice Hasturk, Thomas Van Dyke, Marinella Holzhausen, Alpdogan Kantarci","doi":"10.1080/20002297.2024.2376462","DOIUrl":null,"url":null,"abstract":"<p><strong>Background: </strong>Gingipains are important virulence factors present in Porphyromonas gingivalis. Arginine-specific gingipains (RgpA and RgpB) are critically associated with increased proteolytic activity and immune system dysfunction, including neutrophilic activity. In this study, we assessed the impact of gingipains (RgpA and RgpB) on neutrophil function.</p><p><strong>Methods: </strong>Peripheral blood samples were obtained; neutrophils were isolated and incubated with P. gingivalis A7436, W50, and the double RgpA/RgpB double knockout mutant E8 at MOI 20 for 2 hours. Neutrophil viability was assessed by Sytox staining. Phagocytic capacity and apoptosis were measured by flow cytometry. Superoxide release was measured by superoxide dismutase and cytochrome c reduction assay. Gene expression of TLR2, p47-phox, p67-phox, and P2 × 7was measured by qPCR. Inflammatory cytokine and chemokine production was measured by IL-1β, IL-8, RANTES, and TNF-α in cell supernatants.</p><p><strong>Results: </strong>Neutrophil TLR2 gene expression was reduced in the absence of RgpA/RgpB (<i>p</i> < 0.05), while superoxide production was not significantly impacted. RgpA/RgpB<sup>-/-</sup> significantly impaired neutrophil phagocytic function (<i>p</i> < 0.05) and increased TNF-α production when compared with the wild-type control (<i>p</i> < 0.05). Neutrophil apoptosis was not altered when exposed to RgpA/RgpB<sup>-/-</sup> E8 (<i>p</i> > 0.05).</p><p><strong>Conclusion: </strong>These data suggest that arginine-specific gingipains (RgpA/RgpB) can modulate neutrophil responses against <i>P. gingivalis</i> infection.</p>","PeriodicalId":16598,"journal":{"name":"Journal of Oral Microbiology","volume":"16 1","pages":"2376462"},"PeriodicalIF":3.7000,"publicationDate":"2024-07-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11234918/pdf/","citationCount":"0","resultStr":"{\"title\":\"Arginine-specific gingipains (RgpA/RgpB) knockdown modulates neutrophil machinery.\",\"authors\":\"Vanessa Tubero Euzebio Alves, Tomaz Alves, Emanuel Silva Rovai, Hatice Hasturk, Thomas Van Dyke, Marinella Holzhausen, Alpdogan Kantarci\",\"doi\":\"10.1080/20002297.2024.2376462\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Background: </strong>Gingipains are important virulence factors present in Porphyromonas gingivalis. Arginine-specific gingipains (RgpA and RgpB) are critically associated with increased proteolytic activity and immune system dysfunction, including neutrophilic activity. In this study, we assessed the impact of gingipains (RgpA and RgpB) on neutrophil function.</p><p><strong>Methods: </strong>Peripheral blood samples were obtained; neutrophils were isolated and incubated with P. gingivalis A7436, W50, and the double RgpA/RgpB double knockout mutant E8 at MOI 20 for 2 hours. Neutrophil viability was assessed by Sytox staining. Phagocytic capacity and apoptosis were measured by flow cytometry. Superoxide release was measured by superoxide dismutase and cytochrome c reduction assay. Gene expression of TLR2, p47-phox, p67-phox, and P2 × 7was measured by qPCR. Inflammatory cytokine and chemokine production was measured by IL-1β, IL-8, RANTES, and TNF-α in cell supernatants.</p><p><strong>Results: </strong>Neutrophil TLR2 gene expression was reduced in the absence of RgpA/RgpB (<i>p</i> < 0.05), while superoxide production was not significantly impacted. RgpA/RgpB<sup>-/-</sup> significantly impaired neutrophil phagocytic function (<i>p</i> < 0.05) and increased TNF-α production when compared with the wild-type control (<i>p</i> < 0.05). Neutrophil apoptosis was not altered when exposed to RgpA/RgpB<sup>-/-</sup> E8 (<i>p</i> > 0.05).</p><p><strong>Conclusion: </strong>These data suggest that arginine-specific gingipains (RgpA/RgpB) can modulate neutrophil responses against <i>P. gingivalis</i> infection.</p>\",\"PeriodicalId\":16598,\"journal\":{\"name\":\"Journal of Oral Microbiology\",\"volume\":\"16 1\",\"pages\":\"2376462\"},\"PeriodicalIF\":3.7000,\"publicationDate\":\"2024-07-09\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11234918/pdf/\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of Oral Microbiology\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://doi.org/10.1080/20002297.2024.2376462\",\"RegionNum\":2,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"2024/1/1 0:00:00\",\"PubModel\":\"eCollection\",\"JCR\":\"Q2\",\"JCRName\":\"MICROBIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of Oral Microbiology","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.1080/20002297.2024.2376462","RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2024/1/1 0:00:00","PubModel":"eCollection","JCR":"Q2","JCRName":"MICROBIOLOGY","Score":null,"Total":0}
Background: Gingipains are important virulence factors present in Porphyromonas gingivalis. Arginine-specific gingipains (RgpA and RgpB) are critically associated with increased proteolytic activity and immune system dysfunction, including neutrophilic activity. In this study, we assessed the impact of gingipains (RgpA and RgpB) on neutrophil function.
Methods: Peripheral blood samples were obtained; neutrophils were isolated and incubated with P. gingivalis A7436, W50, and the double RgpA/RgpB double knockout mutant E8 at MOI 20 for 2 hours. Neutrophil viability was assessed by Sytox staining. Phagocytic capacity and apoptosis were measured by flow cytometry. Superoxide release was measured by superoxide dismutase and cytochrome c reduction assay. Gene expression of TLR2, p47-phox, p67-phox, and P2 × 7was measured by qPCR. Inflammatory cytokine and chemokine production was measured by IL-1β, IL-8, RANTES, and TNF-α in cell supernatants.
Results: Neutrophil TLR2 gene expression was reduced in the absence of RgpA/RgpB (p < 0.05), while superoxide production was not significantly impacted. RgpA/RgpB-/- significantly impaired neutrophil phagocytic function (p < 0.05) and increased TNF-α production when compared with the wild-type control (p < 0.05). Neutrophil apoptosis was not altered when exposed to RgpA/RgpB-/- E8 (p > 0.05).
Conclusion: These data suggest that arginine-specific gingipains (RgpA/RgpB) can modulate neutrophil responses against P. gingivalis infection.
期刊介绍：
As the first Open Access journal in its field, the Journal of Oral Microbiology aims to be an influential source of knowledge on the aetiological agents behind oral infectious diseases. The journal is an international forum for original research on all aspects of ''oral health''. Articles which seek to understand ''oral health'' through exploration of the pathogenesis, virulence, host-parasite interactions, and immunology of oral infections are of particular interest. However, the journal also welcomes work that addresses the global agenda of oral infectious diseases and articles that present new strategies for treatment and prevention or improvements to existing strategies.
Topics: ''oral health'', microbiome, genomics, host-pathogen interactions, oral infections, aetiologic agents, pathogenesis, molecular microbiology systemic diseases, ecology/environmental microbiology, treatment, diagnostics, epidemiology, basic oral microbiology, and taxonomy/systematics.
Article types: original articles, notes, review articles, mini-reviews and commentaries
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