Lack of Functional P110δ Affects Expression of Activation Marker CD80 but Does Not Influence Functions of Neutrophils
Neutrophils are specialized immune cells that are essential constituents of the innate immune response. They defend the organism against pathogens through various mechanisms. It was reported that phosphatidylinositols are key players in neutrophil functions, especially in the activity of class-I pho...
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MDPI AG
2022-06-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/23/12/6361 |
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| author | Aneta Manda-Handzlik Agnieszka Mroczek Weronika Kuźmicka Adrianna Cieloch Zuzanna Homoncik Angelika Muchowicz Urszula Demkow Małgorzata Wachowska |
| author_facet | Aneta Manda-Handzlik Agnieszka Mroczek Weronika Kuźmicka Adrianna Cieloch Zuzanna Homoncik Angelika Muchowicz Urszula Demkow Małgorzata Wachowska |
| author_sort | Aneta Manda-Handzlik |
| collection | DOAJ |
| description | Neutrophils are specialized immune cells that are essential constituents of the innate immune response. They defend the organism against pathogens through various mechanisms. It was reported that phosphatidylinositols are key players in neutrophil functions, especially in the activity of class-I phosphoinositide 3-kinases (PI3Ks). P110δ, one of the PI3K subunits, is mostly expressed in immune cells, and its activity plays an important role in inflammatory responses. The aim of this study was to investigate the role of p110δ in neutrophil antimicrobial functions, activation status and cytokine production. To this end, we used bone marrow and splenic neutrophils isolated from a murine model expressing catalytically inactive p110δ<sup>D910A/D910A</sup>. The level of phagocytosis and degranulation, the expressions of activation markers and cytokine production were determined by flow cytometry. ROS generation and NET release were assessed by fluorometry and fluorescent microscopy. We observed a significantly higher percentage of CD80-positive cells among the splenic granulocytes and found granulocytes subpopulations of differing phenotypes between WT and p110δ<sup>D910A/D910A</sup> mice by multiparametric tSNE analysis. Moreover, we detected some differences in the expressions of activation markers, intracellular production of cytokines and bacterial killing. However, we did not observe any alterations in the selected neutrophil functions in p110δ mutant mice. Altogether, our data suggest that the catalytic p110 subunit(s), other than p110δ, is a key player in most neutrophil functions in mice. A follow-up study to correlate these in vitro results with in vivo observations is highly recommended. |
| first_indexed | 2024-03-09T23:37:25Z |
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| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-09T23:37:25Z |
| publishDate | 2022-06-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-d4b450fcac3d4923bf7cc0b787a12d612023-11-23T16:59:22ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-06-012312636110.3390/ijms23126361Lack of Functional P110δ Affects Expression of Activation Marker CD80 but Does Not Influence Functions of NeutrophilsAneta Manda-Handzlik0Agnieszka Mroczek1Weronika Kuźmicka2Adrianna Cieloch3Zuzanna Homoncik4Angelika Muchowicz5Urszula Demkow6Małgorzata Wachowska7Department of Laboratory Diagnostics and Clinical Immunology of Developmental Age, Medical University of Warsaw, Zwirki i Wigury 63a Street, 02-091 Warsaw, PolandDepartment of Laboratory Diagnostics and Clinical Immunology of Developmental Age, Medical University of Warsaw, Zwirki i Wigury 63a Street, 02-091 Warsaw, PolandDepartment of Laboratory Diagnostics and Clinical Immunology of Developmental Age, Medical University of Warsaw, Zwirki i Wigury 63a Street, 02-091 Warsaw, PolandDepartment of Laboratory Diagnostics and Clinical Immunology of Developmental Age, Medical University of Warsaw, Zwirki i Wigury 63a Street, 02-091 Warsaw, PolandDepartment of Laboratory Diagnostics and Clinical Immunology of Developmental Age, Medical University of Warsaw, Zwirki i Wigury 63a Street, 02-091 Warsaw, PolandDepartment of Immunology, Medical University of Warsaw, Jana Nielubowicza 5 Street, 02-097 Warsaw, PolandDepartment of Laboratory Diagnostics and Clinical Immunology of Developmental Age, Medical University of Warsaw, Zwirki i Wigury 63a Street, 02-091 Warsaw, PolandDepartment of Laboratory Diagnostics and Clinical Immunology of Developmental Age, Medical University of Warsaw, Zwirki i Wigury 63a Street, 02-091 Warsaw, PolandNeutrophils are specialized immune cells that are essential constituents of the innate immune response. They defend the organism against pathogens through various mechanisms. It was reported that phosphatidylinositols are key players in neutrophil functions, especially in the activity of class-I phosphoinositide 3-kinases (PI3Ks). P110δ, one of the PI3K subunits, is mostly expressed in immune cells, and its activity plays an important role in inflammatory responses. The aim of this study was to investigate the role of p110δ in neutrophil antimicrobial functions, activation status and cytokine production. To this end, we used bone marrow and splenic neutrophils isolated from a murine model expressing catalytically inactive p110δ<sup>D910A/D910A</sup>. The level of phagocytosis and degranulation, the expressions of activation markers and cytokine production were determined by flow cytometry. ROS generation and NET release were assessed by fluorometry and fluorescent microscopy. We observed a significantly higher percentage of CD80-positive cells among the splenic granulocytes and found granulocytes subpopulations of differing phenotypes between WT and p110δ<sup>D910A/D910A</sup> mice by multiparametric tSNE analysis. Moreover, we detected some differences in the expressions of activation markers, intracellular production of cytokines and bacterial killing. However, we did not observe any alterations in the selected neutrophil functions in p110δ mutant mice. Altogether, our data suggest that the catalytic p110 subunit(s), other than p110δ, is a key player in most neutrophil functions in mice. A follow-up study to correlate these in vitro results with in vivo observations is highly recommended.https://www.mdpi.com/1422-0067/23/12/6361degranulationneutrophilsneutrophil extracellular trapsp110δ subunitphagocytosisphosphoinositide 3-kinase |
| spellingShingle | Aneta Manda-Handzlik Agnieszka Mroczek Weronika Kuźmicka Adrianna Cieloch Zuzanna Homoncik Angelika Muchowicz Urszula Demkow Małgorzata Wachowska Lack of Functional P110δ Affects Expression of Activation Marker CD80 but Does Not Influence Functions of Neutrophils International Journal of Molecular Sciences degranulation neutrophils neutrophil extracellular traps p110δ subunit phagocytosis phosphoinositide 3-kinase |
| title | Lack of Functional P110δ Affects Expression of Activation Marker CD80 but Does Not Influence Functions of Neutrophils |
| title_full | Lack of Functional P110δ Affects Expression of Activation Marker CD80 but Does Not Influence Functions of Neutrophils |
| title_fullStr | Lack of Functional P110δ Affects Expression of Activation Marker CD80 but Does Not Influence Functions of Neutrophils |
| title_full_unstemmed | Lack of Functional P110δ Affects Expression of Activation Marker CD80 but Does Not Influence Functions of Neutrophils |
| title_short | Lack of Functional P110δ Affects Expression of Activation Marker CD80 but Does Not Influence Functions of Neutrophils |
| title_sort | lack of functional p110δ affects expression of activation marker cd80 but does not influence functions of neutrophils |
| topic | degranulation neutrophils neutrophil extracellular traps p110δ subunit phagocytosis phosphoinositide 3-kinase |
| url | https://www.mdpi.com/1422-0067/23/12/6361 |
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