The Differential Reactive Oxygen Species Production of Tear Neutrophils in Response to Various Stimuli In Vitro
A large number of polymorphonuclear neutrophils (PMNs) invade the ocular surface during prolonged eye closure (sleep); these leukocytes are commonly referred as tear PMNs. PMNs contribute to homeostasis and possess an arsenal of inflammatory mediators to protect against pathogens and foreign materia...
| Main Authors: | , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2021-11-01
|
| Series: | International Journal of Molecular Sciences |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1422-0067/22/23/12899 |
| _version_ | 1797507726543159296 |
|---|---|
| author | Yutong Jin Brian Dixon Lyndon Jones Maud Gorbet |
| author_facet | Yutong Jin Brian Dixon Lyndon Jones Maud Gorbet |
| author_sort | Yutong Jin |
| collection | DOAJ |
| description | A large number of polymorphonuclear neutrophils (PMNs) invade the ocular surface during prolonged eye closure (sleep); these leukocytes are commonly referred as tear PMNs. PMNs contribute to homeostasis and possess an arsenal of inflammatory mediators to protect against pathogens and foreign materials. This study examined the ability of tear PMNs to generate reactive oxygen species (ROS), an essential killing mechanism for PMNs which can lead to oxidative stress and imbalance. Cells were collected after sleep from healthy participants using a gentle eye wash. ROS production in stimulated (phorbol-12-myristate-13-acetate (PMA), lipopolysaccharides (LPS) or N-Formylmethionyl-leucyl-phenylalanine (fMLP)) and unstimulated tear PMNs was measured using luminol-enhanced chemiluminescence for 60 min. A high level of constitutive/spontaneous ROS production was observed in tear PMNs in the absence of any stimulus. While tear PMNs were able to produce ROS in response to PMA, they failed to appropriately respond to LPS and fMLP, although fMLP-stimulated tear PMNs generated ROS extracellularly in the first three minutes. Higher ROS generation was observed in isolated tear PMNs which may be due to priming from the magnetic bead cell separation system. The differential responses of tear PMNs in ROS generation provide further evidence of their potential inflammatory roles in ocular complications involving oxidative stress. |
| first_indexed | 2024-03-10T04:53:33Z |
| format | Article |
| id | doaj.art-a3e0437c1b7242228474dd5c3ebea901 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T04:53:33Z |
| publishDate | 2021-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-a3e0437c1b7242228474dd5c3ebea9012023-11-23T02:29:56ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672021-11-0122231289910.3390/ijms222312899The Differential Reactive Oxygen Species Production of Tear Neutrophils in Response to Various Stimuli In VitroYutong Jin0Brian Dixon1Lyndon Jones2Maud Gorbet3School of Optometry and Vision Science, University of Waterloo, Waterloo, ON N2L 3G1, CanadaDepartment of Biology, University of Waterloo, Waterloo, ON N2L 3G1, CanadaSchool of Optometry and Vision Science, University of Waterloo, Waterloo, ON N2L 3G1, CanadaSchool of Optometry and Vision Science, University of Waterloo, Waterloo, ON N2L 3G1, CanadaA large number of polymorphonuclear neutrophils (PMNs) invade the ocular surface during prolonged eye closure (sleep); these leukocytes are commonly referred as tear PMNs. PMNs contribute to homeostasis and possess an arsenal of inflammatory mediators to protect against pathogens and foreign materials. This study examined the ability of tear PMNs to generate reactive oxygen species (ROS), an essential killing mechanism for PMNs which can lead to oxidative stress and imbalance. Cells were collected after sleep from healthy participants using a gentle eye wash. ROS production in stimulated (phorbol-12-myristate-13-acetate (PMA), lipopolysaccharides (LPS) or N-Formylmethionyl-leucyl-phenylalanine (fMLP)) and unstimulated tear PMNs was measured using luminol-enhanced chemiluminescence for 60 min. A high level of constitutive/spontaneous ROS production was observed in tear PMNs in the absence of any stimulus. While tear PMNs were able to produce ROS in response to PMA, they failed to appropriately respond to LPS and fMLP, although fMLP-stimulated tear PMNs generated ROS extracellularly in the first three minutes. Higher ROS generation was observed in isolated tear PMNs which may be due to priming from the magnetic bead cell separation system. The differential responses of tear PMNs in ROS generation provide further evidence of their potential inflammatory roles in ocular complications involving oxidative stress.https://www.mdpi.com/1422-0067/22/23/12899tear neutrophilsocular stressoxidative burstreactive oxygen speciesNADPH oxidaseintracellular pathway |
| spellingShingle | Yutong Jin Brian Dixon Lyndon Jones Maud Gorbet The Differential Reactive Oxygen Species Production of Tear Neutrophils in Response to Various Stimuli In Vitro International Journal of Molecular Sciences tear neutrophils ocular stress oxidative burst reactive oxygen species NADPH oxidase intracellular pathway |
| title | The Differential Reactive Oxygen Species Production of Tear Neutrophils in Response to Various Stimuli In Vitro |
| title_full | The Differential Reactive Oxygen Species Production of Tear Neutrophils in Response to Various Stimuli In Vitro |
| title_fullStr | The Differential Reactive Oxygen Species Production of Tear Neutrophils in Response to Various Stimuli In Vitro |
| title_full_unstemmed | The Differential Reactive Oxygen Species Production of Tear Neutrophils in Response to Various Stimuli In Vitro |
| title_short | The Differential Reactive Oxygen Species Production of Tear Neutrophils in Response to Various Stimuli In Vitro |
| title_sort | differential reactive oxygen species production of tear neutrophils in response to various stimuli in vitro |
| topic | tear neutrophils ocular stress oxidative burst reactive oxygen species NADPH oxidase intracellular pathway |
| url | https://www.mdpi.com/1422-0067/22/23/12899 |
| work_keys_str_mv | AT yutongjin thedifferentialreactiveoxygenspeciesproductionoftearneutrophilsinresponsetovariousstimuliinvitro AT briandixon thedifferentialreactiveoxygenspeciesproductionoftearneutrophilsinresponsetovariousstimuliinvitro AT lyndonjones thedifferentialreactiveoxygenspeciesproductionoftearneutrophilsinresponsetovariousstimuliinvitro AT maudgorbet thedifferentialreactiveoxygenspeciesproductionoftearneutrophilsinresponsetovariousstimuliinvitro AT yutongjin differentialreactiveoxygenspeciesproductionoftearneutrophilsinresponsetovariousstimuliinvitro AT briandixon differentialreactiveoxygenspeciesproductionoftearneutrophilsinresponsetovariousstimuliinvitro AT lyndonjones differentialreactiveoxygenspeciesproductionoftearneutrophilsinresponsetovariousstimuliinvitro AT maudgorbet differentialreactiveoxygenspeciesproductionoftearneutrophilsinresponsetovariousstimuliinvitro |