395887a7979919af7da22bef4aa7c2a6 302129.pdf 00f6276a2575c65887bbb5ced03361c397f48fc6 302129.pdf 1893365afd46487677e723fb5bcb8488f0a0e1bcd9ccfafe44e50a77befbc8eb 302129.pdf Title: Levels of Lysozyme and SLPI in Bronchoalveolar Lavage: Exploring Their Role in Interstitial Lung Disease Subject: Interstitial lung diseases (ILDs) are characterized by inflammation or fibrosis of the pulmonary parenchyma. Despite the involvement of immune cells and soluble mediators in pulmonary fibrosis, the influence of antimicrobial peptides (AMPs) remains underexplored. These effector molecules display a range of activities, which include immunomodulation and wound repair. Here, we investigate the role of AMPs in the development of fibrosis in ILD. We compare the concentration of different AMPs and different cytokines in 46 fibrotic (F-ILD) and 17 non-fibrotic (NF-ILD) patients by ELISA and using peripheral blood mononuclear cells from in vitro stimulation in the presence of lysozyme or secretory leukocyte protease inhibitor (SLPI) from 10 healthy donors. We observed that bronchoalveolar lavage (BAL) levels of AMPs were decreased in F-ILD patients (lysozyme: p < 0.001; SLPI: p < 0.001; LL-37: p < 0.001; lactoferrin: p = 0.47) and were negatively correlated with levels of TGF- (lysozyme: p = 0.02; SLPI: p < 0.001) and IL-17 (lysozyme: p < 0.001; SLPI: p < 0.001). We observed that lysozyme increased the percentage of CD86+ macrophages (p < 0.001) and the production of TNF- (p < 0.001). We showed that lysozyme and SLPI were associated with clinical parameters (lysozyme: p < 0.001; SLPI: p < 0.001) and disease progression (lysozyme: p < 0.001; SLPI: p = 0.01). These results suggest that AMPs may play an important role in the anti-fibrotic response, regulating the effect of pro-fibrotic cytokines. In addition, levels of lysozyme in BAL may be a potential biomarker to predict the progression in F-ILD patients. Keywords: fibrosis; AMPs; immune cells; lysozyme; SLPI Author: Rubén Osuna-Gómez, Maria Mulet, Silvia Barril, Elisabet Cantó, Paloma Millan-Billi, Ana Pardessus, David de la Rosa-Carrillo, Diego Castillo and Silvia Vidal Creator: LaTeX with hyperref Producer: pdfTeX-1.40.25 CreationDate: Mon Apr 15 10:39:21 2024 CEST ModDate: Mon Apr 15 10:49:44 2024 CEST Custom Metadata: no Metadata Stream: no 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: 2980016 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- HJOMHV+VnURWPalladioL-Bold Type 1 Custom yes yes yes 10 0 VVSYYB+EURM10 Type 1 Builtin yes yes yes 16 0 ITSABZ+URWPalladioL-Bold Type 1 Custom yes yes yes 21 0 YFOKPG+URWPalladioL-Roma Type 1 Custom yes yes yes 27 0 WSZDKR+URWPalladioL-Ital Type 1 Custom yes yes yes 32 0 SQEGTJ+VnURWPalladioL Type 1 Custom yes yes yes 37 0 IRWHZX+TimesNewRomanPSMT Type 1C WinAnsi yes yes no 53 0 CKUPDE+TimesNewRomanPS-BoldItalicMT Type 1C WinAnsi yes yes no 56 0 WTAUCZ+CMSY10 Type 1 Builtin yes yes yes 73 0 OEGNEB+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 144 0 OEGNEC+PalatinoLinotype CID TrueType Identity-H yes yes yes 147 0 OEGNED+PalatinoLinotype TrueType WinAnsi yes yes no 153 0 OEGNFE+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 156 0 OEGNLD+PalatinoLinotype TrueType MacRoman yes yes no 159 0 OEGNEB+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 256 0 OEGNED+PalatinoLinotype TrueType WinAnsi yes yes no 259 0 OEGNFE+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 262 0 OEGNLD+PalatinoLinotype TrueType MacRoman yes yes no 265 0 OEGNEB+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 356 0 OEGNEC+PalatinoLinotype CID TrueType Identity-H yes yes yes 359 0 OEGNED+PalatinoLinotype TrueType WinAnsi yes yes no 365 0 OEGNFE+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 368 0 OEGNLD+PalatinoLinotype TrueType MacRoman yes yes no 371 0 OEHKBI+PalatinoLinotype,Italic TrueType MacRoman yes yes no 374 0 OEGNEB+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 417 0 OEGNEC+PalatinoLinotype CID TrueType Identity-H yes yes yes 420 0 OEGNED+PalatinoLinotype TrueType WinAnsi yes yes no 426 0 OEGNFE+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 429 0 OEGNEB+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 489 0 OEGNEC+PalatinoLinotype CID TrueType Identity-H yes yes yes 492 0 OEGNED+PalatinoLinotype TrueType WinAnsi yes yes no 498 0 OEGNFE+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 501 0 OEGNLD+PalatinoLinotype TrueType MacRoman yes yes no 504 0 MPUDFL+PazoMath Type 1 Builtin yes yes yes 535 0 OEGNEB+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 581 0 OEGNEC+PalatinoLinotype CID TrueType Identity-H yes yes yes 584 0 OEGNED+PalatinoLinotype TrueType WinAnsi yes yes no 590 0 OEGNFE+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 593 0 OEGNLD+PalatinoLinotype TrueType MacRoman yes yes no 596 0 DUJUUF+PazoMath-Italic Type 1 Builtin yes yes yes 631 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2024-11-07 02:43:43 CET RepresentationInformation: 302129.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2024-11-06 14:08:41 CET Size: 2980016 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 857 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: section.1 Item: Title: Results Destination: section.2 Children: Item: Title: Population Destination: subsection.2.1 Item: Title: BAL Lysozyme, SLPI, and LL-37 Were Decreased in F-ILD Patients and Associated with Clinical Features Destination: subsection.2.2 Item: Title: Association of Lysozyme and SLPI with Pro- and Anti-Inflammatory Cytokines in the BAL of ILD Patients Destination: subsection.2.3 Item: Title: Lysozyme and SLPI Increased CD86+ Macrophages and Decreased IL-17 Production In Vitro Destination: subsection.2.4 Item: Title: Lysozyme and SLPI Levels Were Associated with Clinical Disease Progression Destination: subsection.2.5 Item: Title: Discussion Destination: section.3 Item: Title: Materials and Methods Destination: section.4 Children: Item: Title: Patients and Design Destination: subsection.4.1 Item: Title: Processing of BAL Destination: subsection.4.2 Item: Title: Cell Culture and Phenotyping by Flow Cytometry Destination: subsection.4.3 Item: Title: Determination of AMPs, IL-10, TGF-, TNF-, and IL-17A in BAL and Culture Supernatants Destination: subsection.4.4 Item: Title: Statistics Destination: subsection.4.5 Item: Title: Conclusions Destination: section.5 Item: Title: References Destination: appendix.A. Info: Title: Levels of Lysozyme and SLPI in Bronchoalveolar Lavage: Exploring Their Role in Interstitial Lung Disease Author: Rubén Osuna-Gómez, Maria Mulet, Silvia Barril, Elisabet Cantó, Paloma Millan-Billi, Ana Pardessus, David de la Rosa-Carrillo, Diego Castillo and Silvia Vidal Subject: Interstitial lung diseases (ILDs) are characterized by inflammation or fibrosis of the pulmonary parenchyma. Despite the involvement of immune cells and soluble mediators in pulmonary fibrosis, the influence of antimicrobial peptides (AMPs) remains underexplored. These effector molecules display a range of activities, which include immunomodulation and wound repair. Here, we investigate the role of AMPs in the development of fibrosis in ILD. We compare the concentration of different AMPs and different cytokines in 46 fibrotic (F-ILD) and 17 non-fibrotic (NF-ILD) patients by ELISA and using peripheral blood mononuclear cells from in vitro stimulation in the presence of lysozyme or secretory leukocyte protease inhibitor (SLPI) from 10 healthy donors. We observed that bronchoalveolar lavage (BAL) levels of AMPs were decreased in F-ILD patients (lysozyme: p < 0.001; SLPI: p < 0.001; LL-37: p < 0.001; lactoferrin: p = 0.47) and were negatively correlated with levels of TGF- (lysozyme: p = 0.02; SLPI: p < 0.001) and IL-17 (lysozyme: p < 0.001; SLPI: p < 0.001). We observed that lysozyme increased the percentage of CD86+ macrophages (p < 0.001) and the production of TNF- (p < 0.001). We showed that lysozyme and SLPI were associated with clinical parameters (lysozyme: p < 0.001; SLPI: p < 0.001) and disease progression (lysozyme: p < 0.001; SLPI: p = 0.01). These results suggest that AMPs may play an important role in the anti-fibrotic response, regulating the effect of pro-fibrotic cytokines. In addition, levels of lysozyme in BAL may be a potential biomarker to predict the progression in F-ILD patients. 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