Profiling endogenous airway proteases and antiproteases and modeling proteolytic activation of Influenza HA using in vitro and ex vivo human airway surface liquid samples.
| Title: | Profiling endogenous airway proteases and antiproteases and modeling proteolytic activation of Influenza HA using in vitro and ex vivo human airway surface liquid samples. |
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| Authors: | Brocke, Stephanie A.1 (AUTHOR), Reidel, Boris2 (AUTHOR), Ehre, Camille2 (AUTHOR), Rebuli, Meghan E.1,3,4 (AUTHOR), Robinette, Carole3 (AUTHOR), Schichlein, Kevin D.1 (AUTHOR), Brooks, Christian A.3 (AUTHOR), Jaspers, Ilona1,3,4,5 (AUTHOR) Ilona_jaspers@med.unc.edu |
| Source: | PLoS ONE. 12/31/2024, Vol. 19 Issue 12, p1-27. 27p. |
| Subject Terms: | *EPITHELIAL cell culture, *AIRWAY (Anatomy), *PEPTIDES, *CIGARETTE smoke, *NASAL irrigation, *PROTEOLYTIC enzymes |
| Abstract: | Imbalance of airway proteases and antiproteases has been implicated in diseases such as COPD and environmental exposures including cigarette smoke and ozone. To initiate infection, endogenous proteases are commandeered by respiratory viruses upon encountering the airway epithelium. The airway proteolytic environment likely contains redundant antiproteases and proteases with diverse catalytic mechanisms, however a proteomic profile of these enzymes and inhibitors in airway samples has not been reported. The objective of this study was to first profile extracellular proteases and antiproteases using human airway epithelial cell cultures and ex vivo nasal epithelial lining fluid (NELF) samples. Secondly, we present an optimized method for probing the proteolytic environment of airway surface liquid samples (in vitro and ex vivo) using fluorogenic peptides modeling the cleavage sites of respiratory viruses. We detected 48 proteases in the apical wash of cultured human nasal epithelial cells (HNECs) (n = 6) and 57 in NELF (n = 13) samples from healthy human subjects using mass-spectrometry based proteomics. Additionally, we detected 29 and 48 antiproteases in the HNEC apical washes and NELF, respectively. We observed large interindividual variability in rate of cleavage of an Influenza H1 peptide in the ex vivo clinical samples. Since protease and antiprotease levels have been found to be altered in the airways of smokers, we compared proteolytic cleavage in ex vivo nasal lavage samples from male/female smokers and non-smokers. There was a statistically significant increase in proteolysis of Influenza H1 in NLF from male smokers compared to female smokers. Furthermore, we measured cleavage of the S1/S2 site of SARS-CoV, SARS-CoV-2, and SARS-CoV-2 Delta peptides in various airway samples, suggesting the method could be used for other viruses of public health relevance. This assay presents a direct and efficient method of evaluating the proteolytic environment of human airway samples in assessment of therapeutic treatment, exposure, or underlying disease. [ABSTRACT FROM AUTHOR] |
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| Items | – Name: Title Label: Title Group: Ti Data: Profiling endogenous airway proteases and antiproteases and modeling proteolytic activation of Influenza HA using in vitro and ex vivo human airway surface liquid samples. – Name: Author Label: Authors Group: Au Data: <searchLink fieldCode="AR" term="%22Brocke%2C+Stephanie+A%2E%22">Brocke, Stephanie A.</searchLink><relatesTo>1</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Reidel%2C+Boris%22">Reidel, Boris</searchLink><relatesTo>2</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Ehre%2C+Camille%22">Ehre, Camille</searchLink><relatesTo>2</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Rebuli%2C+Meghan+E%2E%22">Rebuli, Meghan E.</searchLink><relatesTo>1,3,4</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Robinette%2C+Carole%22">Robinette, Carole</searchLink><relatesTo>3</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Schichlein%2C+Kevin+D%2E%22">Schichlein, Kevin D.</searchLink><relatesTo>1</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Brooks%2C+Christian+A%2E%22">Brooks, Christian A.</searchLink><relatesTo>3</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Jaspers%2C+Ilona%22">Jaspers, Ilona</searchLink><relatesTo>1,3,4,5</relatesTo> (AUTHOR)<i> Ilona_jaspers@med.unc.edu</i> – Name: TitleSource Label: Source Group: Src Data: <searchLink fieldCode="JN" term="%22PLoS+ONE%22">PLoS ONE</searchLink>. 12/31/2024, Vol. 19 Issue 12, p1-27. 27p. – Name: Subject Label: Subject Terms Group: Su Data: *<searchLink fieldCode="DE" term="%22EPITHELIAL+cell+culture%22">EPITHELIAL cell culture</searchLink><br />*<searchLink fieldCode="DE" term="%22AIRWAY+%28Anatomy%29%22">AIRWAY (Anatomy)</searchLink><br />*<searchLink fieldCode="DE" term="%22PEPTIDES%22">PEPTIDES</searchLink><br />*<searchLink fieldCode="DE" term="%22CIGARETTE+smoke%22">CIGARETTE smoke</searchLink><br />*<searchLink fieldCode="DE" term="%22NASAL+irrigation%22">NASAL irrigation</searchLink><br />*<searchLink fieldCode="DE" term="%22PROTEOLYTIC+enzymes%22">PROTEOLYTIC enzymes</searchLink> – Name: Abstract Label: Abstract Group: Ab Data: Imbalance of airway proteases and antiproteases has been implicated in diseases such as COPD and environmental exposures including cigarette smoke and ozone. To initiate infection, endogenous proteases are commandeered by respiratory viruses upon encountering the airway epithelium. The airway proteolytic environment likely contains redundant antiproteases and proteases with diverse catalytic mechanisms, however a proteomic profile of these enzymes and inhibitors in airway samples has not been reported. The objective of this study was to first profile extracellular proteases and antiproteases using human airway epithelial cell cultures and ex vivo nasal epithelial lining fluid (NELF) samples. Secondly, we present an optimized method for probing the proteolytic environment of airway surface liquid samples (in vitro and ex vivo) using fluorogenic peptides modeling the cleavage sites of respiratory viruses. We detected 48 proteases in the apical wash of cultured human nasal epithelial cells (HNECs) (n = 6) and 57 in NELF (n = 13) samples from healthy human subjects using mass-spectrometry based proteomics. Additionally, we detected 29 and 48 antiproteases in the HNEC apical washes and NELF, respectively. We observed large interindividual variability in rate of cleavage of an Influenza H1 peptide in the ex vivo clinical samples. Since protease and antiprotease levels have been found to be altered in the airways of smokers, we compared proteolytic cleavage in ex vivo nasal lavage samples from male/female smokers and non-smokers. There was a statistically significant increase in proteolysis of Influenza H1 in NLF from male smokers compared to female smokers. Furthermore, we measured cleavage of the S1/S2 site of SARS-CoV, SARS-CoV-2, and SARS-CoV-2 Delta peptides in various airway samples, suggesting the method could be used for other viruses of public health relevance. This assay presents a direct and efficient method of evaluating the proteolytic environment of human airway samples in assessment of therapeutic treatment, exposure, or underlying disease. [ABSTRACT FROM AUTHOR] – Name: AbstractSuppliedCopyright Label: Group: Ab Data: <i>Copyright of PLoS ONE is the property of Public Library of Science and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract.</i> (Copyright applies to all Abstracts.) |
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| RecordInfo | BibRecord: BibEntity: Identifiers: – Type: doi Value: 10.1371/journal.pone.0306197 Languages: – Code: eng Text: English PhysicalDescription: Pagination: PageCount: 27 StartPage: 1 Subjects: – SubjectFull: EPITHELIAL cell culture Type: general – SubjectFull: AIRWAY (Anatomy) Type: general – SubjectFull: PEPTIDES Type: general – SubjectFull: CIGARETTE smoke Type: general – SubjectFull: NASAL irrigation Type: general – SubjectFull: PROTEOLYTIC enzymes Type: general Titles: – TitleFull: Profiling endogenous airway proteases and antiproteases and modeling proteolytic activation of Influenza HA using in vitro and ex vivo human airway surface liquid samples. Type: main BibRelationships: HasContributorRelationships: – PersonEntity: Name: NameFull: Brocke, Stephanie A. – PersonEntity: Name: NameFull: Reidel, Boris – PersonEntity: Name: NameFull: Ehre, Camille – PersonEntity: Name: NameFull: Rebuli, Meghan E. – PersonEntity: Name: NameFull: Robinette, Carole – PersonEntity: Name: NameFull: Schichlein, Kevin D. – PersonEntity: Name: NameFull: Brooks, Christian A. – PersonEntity: Name: NameFull: Jaspers, Ilona IsPartOfRelationships: – BibEntity: Dates: – D: 31 M: 12 Text: 12/31/2024 Type: published Y: 2024 Identifiers: – Type: issn-print Value: 19326203 Numbering: – Type: volume Value: 19 – Type: issue Value: 12 Titles: – TitleFull: PLoS ONE Type: main |
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