Interleukin-33 Induces Neutrophil Extracellular Trap (NET) Formation and Macrophage Necroptosis via Enhancing Oxidative Stress and Secretion of Proatherogenic Factors in Advanced Atherosclerosis
Interleukin-33 (IL-33) acts as an ‘alarmin’, and its role has been demonstrated in driving immune regulation and inflammation in many human diseases. However, the precise mechanism of action of IL-33 in regulating neutrophil and macrophage functioning is not defined in advanced atherosclerosis (aAT)...
| Main Authors: | , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2022-11-01
|
| Series: | Antioxidants |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2076-3921/11/12/2343 |
| _version_ | 1827642121025224704 |
|---|---|
| author | Manoj Kumar Tembhre Mukesh Kumar Sriwastva Milind Padmakar Hote Shikha Srivastava Priyanka Solanki Shafaque Imran Ramakrishnan Lakshmy Alpana Sharma Kailash Jaiswal Ashish Datt Upadhyay |
| author_facet | Manoj Kumar Tembhre Mukesh Kumar Sriwastva Milind Padmakar Hote Shikha Srivastava Priyanka Solanki Shafaque Imran Ramakrishnan Lakshmy Alpana Sharma Kailash Jaiswal Ashish Datt Upadhyay |
| author_sort | Manoj Kumar Tembhre |
| collection | DOAJ |
| description | Interleukin-33 (IL-33) acts as an ‘alarmin’, and its role has been demonstrated in driving immune regulation and inflammation in many human diseases. However, the precise mechanism of action of IL-33 in regulating neutrophil and macrophage functioning is not defined in advanced atherosclerosis (aAT) patients. Further, the role of IL-33 in neutrophil extracellular trap (NET) formation in aAT and its consequent effect on macrophage function is not known. In the present study, we recruited <i>n</i> = 52 aAT patients and <i>n</i> = 52 control subjects. The neutrophils were isolated from both groups via ficoll/percoll-based density gradient centrifugation. The effect of IL-33 on the NET formation ability of the neutrophils was determined in both groups. Monocytes, isolated via a positive selection method, were used to differentiate them into macrophages from each of the study subjects and were challenged by IL-33-primed NETs, followed by the measurement of oxidative stress by calorimetric assay and the expression of the proinflammatory molecules by quantitative PCR (qPCR). Transcript and protein expression was determined by qPCR and immunofluorescence/ELISA, respectively. The increased expression of IL-33R (ST-2) was observed in the neutrophils, along with an increased serum concentration of IL-33 in aAT compared to the controls. IL-33 exacerbates NET formation via specifically upregulating CD16 expression in aAT. IL-33-primed NETs/neutrophils increased the cellular oxidative stress levels in the macrophages, leading to enhanced macrophage necroptosis and the release of atherogenic factors and matrix metalloproteinases (MMPs) in aAT compared to the controls. These findings suggested a pathogenic effect of the IL-33/ST-2 pathway in aAT patients by exacerbating NET formation and macrophage necroptosis, thereby facilitating the release of inflammatory factors and the release of MMPs that may be critical for the destabilization/rupture of atherosclerotic plaques in aAT. Targeting the IL-33/ST-2-NETs axis may be a promising therapeutic target for preventing plaque instability/rupture and its adverse complications in aAT. |
| first_indexed | 2024-03-09T17:24:08Z |
| format | Article |
| id | doaj.art-8a055628e24c41c8b6afea0143d56e39 |
| institution | Directory Open Access Journal |
| issn | 2076-3921 |
| language | English |
| last_indexed | 2024-03-09T17:24:08Z |
| publishDate | 2022-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Antioxidants |
| spelling | doaj.art-8a055628e24c41c8b6afea0143d56e392023-11-24T12:56:26ZengMDPI AGAntioxidants2076-39212022-11-011112234310.3390/antiox11122343Interleukin-33 Induces Neutrophil Extracellular Trap (NET) Formation and Macrophage Necroptosis via Enhancing Oxidative Stress and Secretion of Proatherogenic Factors in Advanced AtherosclerosisManoj Kumar Tembhre0Mukesh Kumar Sriwastva1Milind Padmakar Hote2Shikha Srivastava3Priyanka Solanki4Shafaque Imran5Ramakrishnan Lakshmy6Alpana Sharma7Kailash Jaiswal8Ashish Datt Upadhyay9Department of Cardiac Biochemistry, All India Institute of Medical Sciences (AIIMS), New Delhi 110029, IndiaBrown Cancer Center, University of Louisville, Louisville, KY 40202, USADepartment of Cardiothoracic & Vascular Surgery, C. T. Centre, AIIMS, New Delhi 110029, IndiaDepartment of Microbiology & Immunology, University of Louisville, Louisville, KY 40202, USADepartment of Cardiac Biochemistry, All India Institute of Medical Sciences (AIIMS), New Delhi 110029, IndiaDepartment of Cardiac Biochemistry, All India Institute of Medical Sciences (AIIMS), New Delhi 110029, IndiaDepartment of Cardiac Biochemistry, All India Institute of Medical Sciences (AIIMS), New Delhi 110029, IndiaDepartment of Biochemistry, AIIMS, New Delhi 110029, IndiaDepartment of Cardiac Biochemistry, All India Institute of Medical Sciences (AIIMS), New Delhi 110029, IndiaClinical Research Unit (CRU), Biostatistics, AIIMS, New Delhi 110029, IndiaInterleukin-33 (IL-33) acts as an ‘alarmin’, and its role has been demonstrated in driving immune regulation and inflammation in many human diseases. However, the precise mechanism of action of IL-33 in regulating neutrophil and macrophage functioning is not defined in advanced atherosclerosis (aAT) patients. Further, the role of IL-33 in neutrophil extracellular trap (NET) formation in aAT and its consequent effect on macrophage function is not known. In the present study, we recruited <i>n</i> = 52 aAT patients and <i>n</i> = 52 control subjects. The neutrophils were isolated from both groups via ficoll/percoll-based density gradient centrifugation. The effect of IL-33 on the NET formation ability of the neutrophils was determined in both groups. Monocytes, isolated via a positive selection method, were used to differentiate them into macrophages from each of the study subjects and were challenged by IL-33-primed NETs, followed by the measurement of oxidative stress by calorimetric assay and the expression of the proinflammatory molecules by quantitative PCR (qPCR). Transcript and protein expression was determined by qPCR and immunofluorescence/ELISA, respectively. The increased expression of IL-33R (ST-2) was observed in the neutrophils, along with an increased serum concentration of IL-33 in aAT compared to the controls. IL-33 exacerbates NET formation via specifically upregulating CD16 expression in aAT. IL-33-primed NETs/neutrophils increased the cellular oxidative stress levels in the macrophages, leading to enhanced macrophage necroptosis and the release of atherogenic factors and matrix metalloproteinases (MMPs) in aAT compared to the controls. These findings suggested a pathogenic effect of the IL-33/ST-2 pathway in aAT patients by exacerbating NET formation and macrophage necroptosis, thereby facilitating the release of inflammatory factors and the release of MMPs that may be critical for the destabilization/rupture of atherosclerotic plaques in aAT. Targeting the IL-33/ST-2-NETs axis may be a promising therapeutic target for preventing plaque instability/rupture and its adverse complications in aAT.https://www.mdpi.com/2076-3921/11/12/2343Interleukin-33NETsnecroptosisadvanced atherosclerosismyeloperoxidases (MPO)macrophages |
| spellingShingle | Manoj Kumar Tembhre Mukesh Kumar Sriwastva Milind Padmakar Hote Shikha Srivastava Priyanka Solanki Shafaque Imran Ramakrishnan Lakshmy Alpana Sharma Kailash Jaiswal Ashish Datt Upadhyay Interleukin-33 Induces Neutrophil Extracellular Trap (NET) Formation and Macrophage Necroptosis via Enhancing Oxidative Stress and Secretion of Proatherogenic Factors in Advanced Atherosclerosis Antioxidants Interleukin-33 NETs necroptosis advanced atherosclerosis myeloperoxidases (MPO) macrophages |
| title | Interleukin-33 Induces Neutrophil Extracellular Trap (NET) Formation and Macrophage Necroptosis via Enhancing Oxidative Stress and Secretion of Proatherogenic Factors in Advanced Atherosclerosis |
| title_full | Interleukin-33 Induces Neutrophil Extracellular Trap (NET) Formation and Macrophage Necroptosis via Enhancing Oxidative Stress and Secretion of Proatherogenic Factors in Advanced Atherosclerosis |
| title_fullStr | Interleukin-33 Induces Neutrophil Extracellular Trap (NET) Formation and Macrophage Necroptosis via Enhancing Oxidative Stress and Secretion of Proatherogenic Factors in Advanced Atherosclerosis |
| title_full_unstemmed | Interleukin-33 Induces Neutrophil Extracellular Trap (NET) Formation and Macrophage Necroptosis via Enhancing Oxidative Stress and Secretion of Proatherogenic Factors in Advanced Atherosclerosis |
| title_short | Interleukin-33 Induces Neutrophil Extracellular Trap (NET) Formation and Macrophage Necroptosis via Enhancing Oxidative Stress and Secretion of Proatherogenic Factors in Advanced Atherosclerosis |
| title_sort | interleukin 33 induces neutrophil extracellular trap net formation and macrophage necroptosis via enhancing oxidative stress and secretion of proatherogenic factors in advanced atherosclerosis |
| topic | Interleukin-33 NETs necroptosis advanced atherosclerosis myeloperoxidases (MPO) macrophages |
| url | https://www.mdpi.com/2076-3921/11/12/2343 |
| work_keys_str_mv | AT manojkumartembhre interleukin33inducesneutrophilextracellulartrapnetformationandmacrophagenecroptosisviaenhancingoxidativestressandsecretionofproatherogenicfactorsinadvancedatherosclerosis AT mukeshkumarsriwastva interleukin33inducesneutrophilextracellulartrapnetformationandmacrophagenecroptosisviaenhancingoxidativestressandsecretionofproatherogenicfactorsinadvancedatherosclerosis AT milindpadmakarhote interleukin33inducesneutrophilextracellulartrapnetformationandmacrophagenecroptosisviaenhancingoxidativestressandsecretionofproatherogenicfactorsinadvancedatherosclerosis AT shikhasrivastava interleukin33inducesneutrophilextracellulartrapnetformationandmacrophagenecroptosisviaenhancingoxidativestressandsecretionofproatherogenicfactorsinadvancedatherosclerosis AT priyankasolanki interleukin33inducesneutrophilextracellulartrapnetformationandmacrophagenecroptosisviaenhancingoxidativestressandsecretionofproatherogenicfactorsinadvancedatherosclerosis AT shafaqueimran interleukin33inducesneutrophilextracellulartrapnetformationandmacrophagenecroptosisviaenhancingoxidativestressandsecretionofproatherogenicfactorsinadvancedatherosclerosis AT ramakrishnanlakshmy interleukin33inducesneutrophilextracellulartrapnetformationandmacrophagenecroptosisviaenhancingoxidativestressandsecretionofproatherogenicfactorsinadvancedatherosclerosis AT alpanasharma interleukin33inducesneutrophilextracellulartrapnetformationandmacrophagenecroptosisviaenhancingoxidativestressandsecretionofproatherogenicfactorsinadvancedatherosclerosis AT kailashjaiswal interleukin33inducesneutrophilextracellulartrapnetformationandmacrophagenecroptosisviaenhancingoxidativestressandsecretionofproatherogenicfactorsinadvancedatherosclerosis AT ashishdattupadhyay interleukin33inducesneutrophilextracellulartrapnetformationandmacrophagenecroptosisviaenhancingoxidativestressandsecretionofproatherogenicfactorsinadvancedatherosclerosis |