Morphological characterization of para‐ and proinflammatory neutrophil phenotypes using transmission electron microscopy

Borenstein, Alon; Fine, Noah; Hassanpour, Siavash; Sun, Cong; Oveisi, Morvarid; Tenenbaum, Howard C.; Glogauer, Michael

doi:10.1111/jre.12595

Cited by 18 publications

(26 citation statements)

References 48 publications

(84 reference statements)

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“…It has been previously shown that not all neutrophils in the oral cavity display similar levels of cell surface CD marker expression (5,11,12). In the healthy oral cavity, neutrophils are in a parainflammatory state, whereas in a diseased cavity, neutrophil phenotypes are changed to a proinflammatory state, with neutrophils in the parain-flammatory and proinflammatory states expressing low levels and high levels of CD markers, respectively (5).…”

Section: Resultsmentioning

confidence: 99%

Novel Assay To Characterize Neutrophil Responses to Oral Biofilms

et al. 2019

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Neutrophils, the most numerous leukocytes, play an important role in maintaining oral health through interactions with oral microbial biofilms. Both neutrophil hyperactivity and the bacterial subversion of neutrophil responses can cause inflammation-mediated tissue damage like that seen in periodontal disease. We describe here an assay that assesses neutrophil activation responses to monospecies biofilm bacteriain vitrobased on the surface expression of cluster of differentiation (CD) markers associated with various neutrophil functions. Most of what we know about neutrophil responses to bacteria is based onin vitroassays that use planktonic bacteria and isolated/preactivated neutrophils, which makes interpretation of the neutrophil responses to bacteria a challenge. An understanding of how neutrophils differentially interact with and respond to commensal and pathogenic oral bacteria is necessary in order to further understand the neutrophil’s role in maintaining oral health and the pathogenesis of periodontal disease. In this study, a flow cytometry-basedin vitroassay was developed to characterize neutrophil activation states based on CD marker expressions in response to oral monospecies bacterial biofilms. Using this approach, changes in CD marker expressions in response to specific prominent oral commensal and pathogenic bacteria were assayed. Several functional assays, including assays for phagocytosis, production of reactive oxygen species, activation of the transcription factor Nrf2, neutrophil extracellular trap formation, and myeloperoxidase release, were also performed to correlate neutrophil function with CD marker expression. Our results demonstrate that neutrophils display bacterial species-specific responses. This assay can be used to characterize how specific biofilms alter specific neutrophil pathways associated with their activation.

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Section: Resultsmentioning

confidence: 99%

Novel Assay To Characterize Neutrophil Responses to Oral Biofilms

et al. 2019

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“…Images of Western blot and nuclear euchromatin fraction were analyzed by Image J (National Institutes of Health, Bethesda, MD, USA) according to a previous study. 22 The number of neutrophil fluorescent spots were counted using Image-Pro Plus 6.0 (Media Cybernetics, Inc., Rockville, MD, USA). Statistical analysis and charting were performed with GraphPad Prism 5 (GraphPad Software, San Diego, CA, USA).…”

Section: Discussionmentioning

confidence: 99%

The Role of Autophagy in the Innate Immune Response to Fungal Keratitis Caused by Aspergillus fumigatus Infection

Cui

Lin

et al. 2020

Invest. Ophthalmol. Vis. Sci.

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Citation: Li C, Li C, Lin J, et al. The role of autophagy in the innate immune response to fungal keratitis caused by Aspergillus fumigatus infection. Invest Ophthalmol Vis Sci. 2020;61(2):25. https://doi.org/10.1167/iovs.61.2.25 PURPOSE.To determine the role of autophagy in the innate immune response to fungal keratitis (FK) caused by Aspergillus fumigatus infection. METHODS.Corneal samples obtained from patients and mice with FK were visualized via transmission electron microscopy (TEM). Autophagy-related proteins LC3B-II, Beclin-1, LAMP-1, and p62 in A. fumigatus-infected corneas of C57BL/6 mice were tested by Western blot. After treatment with autophagy inhibitors 3-methyladenine (3-MA), chloroquine (CQ), or inducer rapamycin, autophagy-related proteins were detected by Western blot. Corneas were photographed with slit lamp microscopy and pathological changes were observed by hematoxylin and eosin staining. Polymorphonuclear neutrophilic leukocytes (PMNs) were assessed by immunofluorescent staining and observed under TEM. The levels of CXCL-1, IL-1β, HMGB1, IL-18, TNF-α, and IL-10 were tested by reverse transcription polymerase chain reaction and Western blot. The quantification of fungal loads was detected and photographed. RESULTS.The accumulation of autophagosomes in corneas of patients and mice with FK was observed with TEM. The expression of LC3B-II, Beclin-1, and LAMP-1 was elevated in corneas after fungal infection, whereas p62 was reduced. Treatment with 3-MA or CQ upregulated clinical scores, pathological changes, and the expression of CXCL-1, IL-1β, HMGB1, IL-18, and TNF-α except IL-10. The morphology of PMNs was changed and PMN recruitment was increased in mice corneas treated with 3-MA or CQ, whereas rapamycin reduced the inflammatory response to keratitis. These results were statistically significant. CONCLUSIONS. A. fumigatus infection increases the expression of autophagy in corneas.Autophagy plays an anti-inflammatory role in the innate immune response to A. fumigatus keratitis.

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“…Circulatory neutrophils, on the other hand, were shown to be activated during gingivitis onset as shown by the markers CD55, CD63, CD11b, and CD66a. Also in 2018, this group further characterized the morphology of the para-inflammatory health-associated phenotype described in 2016, and compared it to the naïve morphology of blood neutrophils as well as to the pro-inflammatory phenotype of oral neutrophils in periodontitis (58). They revealed that pro-inflammatory neutrophils in periodontitis showed less granulation, lighter cytoplasms and higher amounts of nuclear euchromatin, as assessed by electron microscopy, compared to the para-inflammatory oral health neutrophils.…”

Section: Possible Neutrophil Subsets In Periodontal Health and Inflammentioning

confidence: 99%

Neutrophil Subsets in Periodontal Health and Disease: A Mini Review

Hirschfeld

2020

Front. Immunol.

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Neutrophils are amongst the most abundant immune cells within the periodontal tissues and oral cavity. As innate immune cells, they are first line defenders at the tooth-mucosa interface, and can perform an array of different functions. With regard to these, it has been observed over many years that neutrophils are highly heterogeneous in their behavior. Therefore, it has been speculated that neutrophils, similarly to other leukocytes, exist in distinct subsets. Several studies have investigated different markers of neutrophils in oral health and disease in recent years in order to define potential cell subsets and their specific tasks. This research was inspired by recent advancements in other fields of medicine in this field. The aim of this review is to give an overview of the current evidence regarding the existence and presence of neutrophil subsets and their possible functions, specifically in the context of periodontitis, gingivitis, and periodontal health.

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Morphological characterization of para‐ and proinflammatory neutrophil phenotypes using transmission electron microscopy

Cited by 18 publications

References 48 publications

Novel Assay To Characterize Neutrophil Responses to Oral Biofilms

Novel Assay To Characterize Neutrophil Responses to Oral Biofilms

The Role of Autophagy in the Innate Immune Response to Fungal Keratitis Caused by Aspergillus fumigatus Infection

Neutrophil Subsets in Periodontal Health and Disease: A Mini Review

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