0f1a1effaa3ece2bd260ad6576a33d10 302823.pdf 2bf44d57d7ecf734e043e6dde33c8f9e30125dcd 302823.pdf 31a292b7afcee7e9f892f75ca12ffb10a776374209b9a62be65b157f944b9352 302823.pdf Title: The Effects of Interstitial Lung Diseases on Alveolar Extracellular Vesicles Profile: A Multicenter Study Subject: Diagnosis of interstitial lung diseases (ILD) is difficult to perform. Extracellular vesicles (EVs) facilitate cell-to-cell communication, and they are released by a variety of cells. Our goal aimed to investigate EV markers in bronchoalveolar lavage (BAL) from idiopathic pulmonary fibrosis (IPF), sarcoidosis and hypersensitivity pneumonitis (HP) cohorts. ILD patients followed at Siena, Barcelona and Foggia University Hospitals were enrolled. BAL supernatants were used to isolate the EVs. They were characterized by flow cytometry assay through MACSPlex Exsome KIT. The majority of alveolar EV markers were related to the fibrotic damage. CD56, CD105, CD142, CD31 and CD49e were exclusively expressed by alveolar samples from IPF patients, while HP showed only CD86 and CD24. Some EV markers were common between HP and sarcoidosis (CD11c, CD1c, CD209, CD4, CD40, CD44, CD8). Principal component analysis distinguished the three groups based on EV markers with total variance of 60.08%. This study has demonstrated the validity of the flow cytometric method to phenotype and characterize EV surface markers in BAL samples. The two granulomatous diseases, sarcoidosis and HP, cohorts shared alveolar EV markers not revealed in IPF patients. Our findings demonstrated the viability of the alveolar compartment allowing identification of lung-specific markers for IPF and HP. Keywords: extracellular vesicles; interstitial lung diseases; bronchoalveolar lavage; idiopathicpulmonary fibrosis; diagnosis Author: Miriana d’Alessandro, Sara Gangi, Piera Soccio, Elisabet Cantó, Rubén Osuna-Gómez, Laura Bergantini, Paolo Cameli, Gaia Fabbri, Sara Croce, Giulia Scioscia, Giusy Montuori, Matteo Fanetti, Giorgia Moriondo, Fabrizio Mezzasalma, Diego Castillo, Donato Lacedonia, Silvia Vidal and Elena Bargagli Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Fri Feb 17 10:18:34 2023 CET ModDate: Fri Feb 17 10:34:41 2023 CET Custom Metadata: no Metadata Stream: no Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 12 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 3115749 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- XKZCWQ+URWPalladioL-Bold Type 1 Custom yes yes no 10 0 OFKWDN+URWPalladioL-Roma Type 1 Custom yes yes no 15 0 KTNPUB+URWPalladioL-Ital Type 1 Custom yes yes no 19 0 HJOMHV+VnURWPalladioL-Bold Type 1 Custom yes yes no 23 0 IRWHZX+TimesNewRomanPSMT Type 1C WinAnsi yes yes no 42 0 CKUPDE+TimesNewRomanPS-BoldItalicMT Type 1C WinAnsi yes yes no 45 0 QMDDXF+CMSY10 Type 1 Builtin yes yes no 70 0 URYLRZ+URWPalladioL-BoldItal Type 1 Custom yes yes no 74 0 MLPBPF+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 88 0 MLPBPH+PalatinoLinotype TrueType WinAnsi yes yes no 91 0 MLPBPJ+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 94 0 MLPBPF+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 105 0 MLPBPH+PalatinoLinotype TrueType WinAnsi yes yes no 108 0 MLPBPJ+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 111 0 MLPBPF+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 126 0 MLPBPH+PalatinoLinotype TrueType WinAnsi yes yes no 129 0 MLPBPJ+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 132 0 MLPBPF+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 144 0 MLPBPG+PalatinoLinotype CID TrueType Identity-H yes yes yes 147 0 MLPBPH+PalatinoLinotype TrueType WinAnsi yes yes no 153 0 MLPBPJ+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 156 0 BCDEEE+Calibri-Bold TrueType WinAnsi yes yes no 171 0 OVNEXW+EURM10 Type 1 Builtin yes yes no 205 0 YWZDMF+VnURWPalladioL Type 1 Custom yes yes no 215 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2024-11-19 02:45:34 CET RepresentationInformation: 302823.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2024-11-18 10:01:11 CET Size: 3115749 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 402 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: Patients Destination: subsection.2.1 Item: Title: EVs Surface Markers Destination: subsection.2.2 Item: Title: Discussion Destination: section.3 Item: Title: Materials and Methods Destination: section.4 Children: Item: Title: Study Cohort Destination: subsection.4.1 Item: Title: Validation Cohort Destination: subsection.4.2 Item: Title: Pulmonary Function Test Parameters Destination: subsection.4.3 Item: Title: Bronchoscopy Destination: subsection.4.4 Item: Title: EVs Isolation and Characterization Destination: subsection.4.5 Item: Title: Statistical Analysis Destination: subsection.4.6 Item: Title: Conclusions Destination: section.5 Item: Title: References Destination: section.5 Info: Title: The Effects of Interstitial Lung Diseases on Alveolar Extracellular Vesicles Profile: A Multicenter Study Author: Miriana d’Alessandro, Sara Gangi, Piera Soccio, Elisabet Cantó, Rubén Osuna-Gómez, Laura Bergantini, Paolo Cameli, Gaia Fabbri, Sara Croce, Giulia Scioscia, Giusy Montuori, Matteo Fanetti, Giorgia Moriondo, Fabrizio Mezzasalma, Diego Castillo, Donato Lacedonia, Silvia Vidal and Elena Bargagli Subject: Diagnosis of interstitial lung diseases (ILD) is difficult to perform. Extracellular vesicles (EVs) facilitate cell-to-cell communication, and they are released by a variety of cells. Our goal aimed to investigate EV markers in bronchoalveolar lavage (BAL) from idiopathic pulmonary fibrosis (IPF), sarcoidosis and hypersensitivity pneumonitis (HP) cohorts. ILD patients followed at Siena, Barcelona and Foggia University Hospitals were enrolled. BAL supernatants were used to isolate the EVs. They were characterized by flow cytometry assay through MACSPlex Exsome KIT. The majority of alveolar EV markers were related to the fibrotic damage. CD56, CD105, CD142, CD31 and CD49e were exclusively expressed by alveolar samples from IPF patients, while HP showed only CD86 and CD24. Some EV markers were common between HP and sarcoidosis (CD11c, CD1c, CD209, CD4, CD40, CD44, CD8). Principal component analysis distinguished the three groups based on EV markers with total variance of 60.08%. This study has demonstrated the validity of the flow cytometric method to phenotype and characterize EV surface markers in BAL samples. The two granulomatous diseases, sarcoidosis and HP, cohorts shared alveolar EV markers not revealed in IPF patients. Our findings demonstrated the viability of the alveolar compartment allowing identification of lung-specific markers for IPF and HP. 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