887e96af684437a7cf188c26f9a93db1 284180.pdf 2f1f185d5ec40f49157b238178005ca755beda0e 284180.pdf f39119a11784a07bb6cb45672e05b559a60c9416a9b9b830f3e3f4757ecd076b 284180.pdf Title: Influence of Malignant Pleural Fluid from Lung Adenocarcinoma Patients on Neutrophil Response Subject: Malignant pleural effusion (MPE) is a common severe complication of advanced lung adenocarcinoma (LAC). Neutrophils, an essential component of tumor infiltrates, contribute to tumor progression and their counts in MPE have been associated with worse outcome in LAC. This study aimed to evaluate phenotypical and functional changes of neutrophils induced by MPE to determine the influence of MPE immunomodulatory factors in neutrophil response and to find a possible association between neutrophil functions and clinical outcomes. Pleural fluid samples were collected from 47 LAC and 25 heart failure (HF) patients. We measured neutrophil degranulation products by ELISA, oxidative burst capacity and apoptosis by flow cytometry, and NETosis by fluorescence. The concentration of degranulation products was higher in MPE-LAC than in PE-HF. Functionally, neutrophils cultured with MPE-LAC had enhanced survival and neutrophil extracellular trap (NET) formation but had reduced oxidative burst capacity. In MPE, NETosis was positively associated with MMP-9, P-selectin, and sPD-L1 and clinically related to a worse outcome. This is the first study associating NETs with a worse outcome in MPE. Neutrophils likely contribute to tumor progression through the release of NETs, suggesting that they are a potential therapeutic target in LAC. Keywords: malignant pleural fluid; lung adenocarcinoma; neutrophils; NETosis Author: Maria Mulet, Rubén Osuna-Gómez, Carlos Zamora, José M. Porcel, Juan C. Nieto, Lídia Perea, Virginia Pajares, Ana M. Muñoz-Fernandez, Nuria Calvo, Maria Alba Sorolla and Silvia Vidal Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Mon May 23 11:05:11 2022 CEST ModDate: Mon May 23 11:09:17 2022 CEST Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 14 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 1782207 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- HIDLLI+URWPalladioL-Bold Type 1 Custom yes yes no 10 0 KXZIMK+URWPalladioL-Roma Type 1 Custom yes yes no 15 0 CSZMWO+URWPalladioL-Ital Type 1 Custom yes yes no 19 0 XWOCXD+VnURWPalladioL-Bold Type 1 Custom yes yes no 23 0 LCJEVJ+TimesNewRomanPS-BoldItalicMT Type 1C WinAnsi yes yes no 43 0 QMDDXF+CMSY10 Type 1 Builtin yes yes no 68 0 WBVKDJ+URWPalladioL-BoldItal Type 1 Custom yes yes no 72 0 BBSDEQ+EURM10 Type 1 Builtin yes yes no 76 0 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BKPAMC+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 207 0 XEFGQJ+VnURWPalladioL Type 1 Custom yes yes no 224 0 Jhove (Rel. 1.22.1, 2019-04-17) Date: 2023-11-30 02:36:40 CET RepresentationInformation: 284180.pdf ReportingModule: PDF-hul, Rel. 1.12.1 (2019-04-17) LastModified: 2023-11-29 13:19:29 CET Size: 1782207 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 404 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@4d3167f4 Item: Title: Materials and Methods Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@ed9d034 Children: Item: Title: Patients and Pleural Fluid Samples Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@6121c9d6 Item: Title: Phenotyping Neutrophils from Pleural Effusions by Flow Cytometry Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@87f383f Item: Title: Isolation of Neutrophils from Healthy Donors Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@4eb7f003 Item: Title: Induction and Quantification of NETosis in the Presence of Cell-Free Pleural Fluids Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@eafc191 Item: Title: Measurement of Oxidative Burst in the Presence of Cell-Free Pleural Fluids Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@612fc6eb Item: Title: Induction of Neutrophil Apoptosis in the Presence of Cell-Free Pleural Fluids Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1060b431 Item: Title: Determination of Biochemical Parameters and Soluble Mediators in Pleural Fluids Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@612679d6 Item: Title: Statistics Analysis Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@11758f2a Item: Title: Results Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@e720b71 Children: Item: Title: Phenotype of Pleural Neutrophils and Concentration of Neutrophil-Associated Molecules in Pleural Fluids Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1b26f7b2 Item: Title: Effect of Cell-Free Pleural Fluid on Neutrophil Viability and Function Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@491cc5c9 Item: Title: Association between Immunomodulatory Factors in Pleural Fluid and Neutrophil Function Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@74ad1f1f Item: Title: Association between Neutrophil Functions and the Clinical Outcome of LAC Patients Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@6a1aab78 Item: Title: Discussion Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@462d5aee Item: Title: Conclusions Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@69b0fd6f Item: Title: References Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@757942a1 Info: Title: Influence of Malignant Pleural Fluid from Lung Adenocarcinoma Patients on Neutrophil Response Author: Maria Mulet, Rubén Osuna-Gómez, Carlos Zamora, José M. Porcel, Juan C. Nieto, Lídia Perea, Virginia Pajares, Ana M. Muñoz-Fernandez, Nuria Calvo, Maria Alba Sorolla and Silvia Vidal Subject: Malignant pleural effusion (MPE) is a common severe complication of advanced lung adenocarcinoma (LAC). Neutrophils, an essential component of tumor infiltrates, contribute to tumor progression and their counts in MPE have been associated with worse outcome in LAC. This study aimed to evaluate phenotypical and functional changes of neutrophils induced by MPE to determine the influence of MPE immunomodulatory factors in neutrophil response and to find a possible association between neutrophil functions and clinical outcomes. Pleural fluid samples were collected from 47 LAC and 25 heart failure (HF) patients. We measured neutrophil degranulation products by ELISA, oxidative burst capacity and apoptosis by flow cytometry, and NETosis by fluorescence. The concentration of degranulation products was higher in MPE-LAC than in PE-HF. Functionally, neutrophils cultured with MPE-LAC had enhanced survival and neutrophil extracellular trap (NET) formation but had reduced oxidative burst capacity. In MPE, NETosis was positively associated with MMP-9, P-selectin, and sPD-L1 and clinically related to a worse outcome. This is the first study associating NETs with a worse outcome in MPE. Neutrophils likely contribute to tumor progression through the release of NETs, suggesting that they are a potential therapeutic target in LAC. 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