FRINK Linked Data Fragments server

Matches in SemOpenAlex for { <https://semopenalex.org/work/W2924099150> ?p ?o ?g. }

• W2924099150 endingPage "5095" @default.
• W2924099150 startingPage "5095" @default.
• W2924099150 abstract "Abstract Background Mesenchymal stem/stromal cells (MSCs) are a promising treatment for acute graft-versus-host disease and other inflammatory diseases. MSCs express programmed death-ligand 1 (PD-L1) that has been reported to contribute to MSC-mediated immune suppression through T-cell receptor (TCR)-mediated inhibitory signaling. MSCs actively secrete extracellular vesicles (EVs) that have immune modulatory properties resembling their cell of origin. Although the release of EVs by MSCs has been extensively reported, the presence and functional impact of immune inhibitory checkpoint molecules in MSC-derived EVs is still being determined. We sought to determine if PD-L1 and other immune inhibitory checkpoint molecules are enriched in MSC-derived EVs. In this study we report that PD-L1 is enriched in MSC-EVs, and that PD-L1(+) EVs and PD-L1(-) EVs differ in their T cell immune suppressive capacity. Methods Umbilical cords and healthy donor blood samples were collected on separate IRB-approved protocols after written informed consent (HSC#1546/HSC#5929). Umbilical cord Wharton's jelly-MSCs (WJMSC) were isolated from umbilical cords under GMP conditions and characterized by morphology, multiparameter flow cytometry (MPFC), and differentiation capacity. Three clinical-grade WJMSC cell lines were cultured at low passage (2-5) in EV-depleted media. EVs were isolated from the media by differential ultracentrifugation and the 100K pellets enriched with exosomes were characterized by Western blot (WB), nanoparticle tracking analysis (NTA), and electron microscopy (EM). To further identify EV-enriched immune proteins, the WJMSC and EV proteomes were characterized by Tandem Mass Tag (TMT) labeling and LC MS/MS, using isobaric labels to compare protein quantities between samples. Gene disruption of PD-L1 was performed through site-specific CRISPR/Cas9 introducing a single cysteine nucleotide deletion in exon 3; the genetic mutation was confirmed by Sanger sequencing. The absence of PD-L1 transcript was confirmed by RT-PCR, and the absence of PD-L1 protein was confirmed by WB, immunofluorescence, MPFC, and IFNγ-response. PBMC were isolated from healthy donor blood samples; T cells were activated by CD3/CD28 beads and induced CD154 was measured on CD4(+) T cells by MPFC. Alternatively, CMV-experienced T cells were activated with CMV-pp65 peptide and induced IFNγ was measured on CD8(+) T cells by MPFC. Activated T cells were treated with PD-L1(+) and PD-L1(-) WJMSCs or their derived EVs. Phospho-Zap-70 (Tyr319)/Syk (Tyr352) MPFC was used to analyze T-cell receptor (TCR)-mediated signaling. Results WJMSCs were CD73(+), CD90(+), CD105(+), and CD34(-), CD45(-), CD11b(-), CD19(-) and HLA-DR(-); and inhibited proliferation in a mixed lymphocyte reaction. WJMSCs-derived EVs were cup-shaped 100-200nm nanoparticles by EM and were CD9(+), CD81(+), CD73(+), CD90(+), and CD105(+) by WB. Quantitative proteomic profiling revealed a total of 1,634 proteins across WJMSC-derived EV preparations. Proteins that were highly enriched in MSC-derived EVs and that were classified under negative regulation of lymphocyte activation (Gene Ontology Biological Processes; GOBP) included PDL1, CD276, DLG1, TNR6, ERBB2, TGFβ1, LYN, and HMGB3. Both WJMSCs and EVs suppressed CD4(+) T cell activation by CD3/CD28 beads and CD8(+) T cell activation by CMV peptide. CRISPR/Cas9 edited WJMSC exhibited normal morphology and differentiation characteristics. PD-L1 gene disruption efficiently reduced PD-L1 protein expression on WJMSC and their derived EVs, and reduced the ability of both to suppress T cell activation. Similarly, blocking PD-L1 with a specific internalization antibody decreased WJMSC and EV-mediated T cell activation. Conclusion Proteomic analyses reveal that PD-L1 is enriched in WJMSC-derived EVs. CRISPR/Cas9-mediated disruption of PD-L1 in WJMSCs alone did not affect the release of EVs in vitro. Gene-edited WJMSCs stably generate PD-L1(-) EVs. The PD-L1(-) EVs generated from PD-L1(-) WJMSC did not suppress T cell activation in co-culture. These findings indicate that PD-L1 is enriched in WJMSC EVs and is biologically active. WJMSC-EVs can deliver membrane-bound PD-L1 to T cells, resulting in attenuated TCR signaling. Disclosures McGuirk: Fresenius Biotech: Research Funding; Bellicum Pharmaceuticals: Research Funding; Astellas Pharma: Research Funding; Pluristem Ltd: Research Funding; Gamida Cell: Research Funding; Kite Pharma: Honoraria, Other: travel accommodations, expenses, speaker ; Novartis Pharmaceuticals Corporation: Honoraria, Other: speaker, Research Funding." @default.
• W2924099150 created "2019-04-01" @default.
• W2924099150 creator A5026868791 @default.
• W2924099150 creator A5030605391 @default.
• W2924099150 creator A5033625395 @default.
• W2924099150 creator A5051905584 @default.
• W2924099150 creator A5052064208 @default.
• W2924099150 creator A5078398693 @default.
• W2924099150 creator A5079800586 @default.
• W2924099150 creator A5086255337 @default.
• W2924099150 creator A5086587597 @default.
• W2924099150 date "2018-11-29" @default.
• W2924099150 modified "2023-10-16" @default.
• W2924099150 title "CRISPR/Cas9-Mediated Disruption of PD-L1 Reduces the T Cell Suppressive Effect of Wharton's Jelly Mesenchymal Stromal Cells and Their Extracellular Vesicles" @default.
• W2924099150 doi "https://doi.org/10.1182/blood-2018-99-116921" @default.
• W2924099150 hasPublicationYear "2018" @default.
• W2924099150 type Work @default.
• W2924099150 sameAs 2924099150 @default.
• W2924099150 citedByCount "0" @default.
• W2924099150 crossrefType "journal-article" @default.
• W2924099150 hasAuthorship W2924099150A5026868791 @default.
• W2924099150 hasAuthorship W2924099150A5030605391 @default.
• W2924099150 hasAuthorship W2924099150A5033625395 @default.
• W2924099150 hasAuthorship W2924099150A5051905584 @default.
• W2924099150 hasAuthorship W2924099150A5052064208 @default.
• W2924099150 hasAuthorship W2924099150A5078398693 @default.
• W2924099150 hasAuthorship W2924099150A5079800586 @default.
• W2924099150 hasAuthorship W2924099150A5086255337 @default.
• W2924099150 hasAuthorship W2924099150A5086587597 @default.
• W2924099150 hasBestOaLocation W29240991501 @default.
• W2924099150 hasConcept C104317684 @default.
• W2924099150 hasConcept C119577978 @default.
• W2924099150 hasConcept C145059251 @default.
• W2924099150 hasConcept C16930146 @default.
• W2924099150 hasConcept C185592680 @default.
• W2924099150 hasConcept C198826908 @default.
• W2924099150 hasConcept C203014093 @default.
• W2924099150 hasConcept C20518536 @default.
• W2924099150 hasConcept C2781261824 @default.
• W2924099150 hasConcept C2908689518 @default.
• W2924099150 hasConcept C502942594 @default.
• W2924099150 hasConcept C55493867 @default.
• W2924099150 hasConcept C86803240 @default.
• W2924099150 hasConcept C8891405 @default.
• W2924099150 hasConcept C95444343 @default.
• W2924099150 hasConceptScore W2924099150C104317684 @default.
• W2924099150 hasConceptScore W2924099150C119577978 @default.
• W2924099150 hasConceptScore W2924099150C145059251 @default.
• W2924099150 hasConceptScore W2924099150C16930146 @default.
• W2924099150 hasConceptScore W2924099150C185592680 @default.
• W2924099150 hasConceptScore W2924099150C198826908 @default.
• W2924099150 hasConceptScore W2924099150C203014093 @default.
• W2924099150 hasConceptScore W2924099150C20518536 @default.
• W2924099150 hasConceptScore W2924099150C2781261824 @default.
• W2924099150 hasConceptScore W2924099150C2908689518 @default.
• W2924099150 hasConceptScore W2924099150C502942594 @default.
• W2924099150 hasConceptScore W2924099150C55493867 @default.
• W2924099150 hasConceptScore W2924099150C86803240 @default.
• W2924099150 hasConceptScore W2924099150C8891405 @default.
• W2924099150 hasConceptScore W2924099150C95444343 @default.
• W2924099150 hasIssue "Supplement 1" @default.
• W2924099150 hasLocation W29240991501 @default.
• W2924099150 hasOpenAccess W2924099150 @default.
• W2924099150 hasPrimaryLocation W29240991501 @default.
• W2924099150 hasRelatedWork W2907368035 @default.
• W2924099150 hasRelatedWork W2924099150 @default.
• W2924099150 hasRelatedWork W2944096190 @default.
• W2924099150 hasRelatedWork W2945013784 @default.
• W2924099150 hasRelatedWork W2955875101 @default.
• W2924099150 hasRelatedWork W2964419615 @default.
• W2924099150 hasRelatedWork W3009964850 @default.
• W2924099150 hasRelatedWork W3139087999 @default.
• W2924099150 hasRelatedWork W3207489770 @default.
• W2924099150 hasRelatedWork W4224750750 @default.
• W2924099150 hasVolume "132" @default.
• W2924099150 isParatext "false" @default.
• W2924099150 isRetracted "false" @default.
• W2924099150 magId "2924099150" @default.
• W2924099150 workType "article" @default.