The human glomerular endothelial cells are potent pro-inflammatory contributors in an in vitro model of lupus nephritis
Identifieur interne : 000345 ( Main/Exploration ); précédent : 000344; suivant : 000346The human glomerular endothelial cells are potent pro-inflammatory contributors in an in vitro model of lupus nephritis
Auteurs : Paraskevi Dimou [Royaume-Uni] ; Rachael D. Wright [Royaume-Uni] ; Kelly L. Budge [Royaume-Uni] ; Angela Midgley [Royaume-Uni] ; Simon C. Satchell [Royaume-Uni] ; Matthew Peak [Royaume-Uni] ; Michael W. Beresford [Royaume-Uni]Source :
- Scientific Reports [ 2045-2322 ] ; 2019.
Abstract
Juvenile-onset lupus nephritis (LN) affects up to 80% of juvenile-onset systemic lupus erythematosus patients (JSLE). As the exact role of human renal glomerular endothelial cells (GEnCs) in LN has not been fully elucidated, the aim of this study was to investigate their involvement in LN. Conditionally immortalised human GEnCs (ciGEnCs) were treated with pro-inflammatory cytokines known to be involved in LN pathogenesis and also with LPS. Secretion and surface expression of pro-inflammatory proteins was quantified via ELISA and flow cytometry. NF-κΒ and STAT-1 activation was investigated via immunofluorescence. Serum samples from JSLE patients and from healthy controls were used to treat ciGEnCs to determine via qRT-PCR potential changes in the mRNA levels of pro-inflammatory genes. Our results identified TNF-α, IL-1β, IL-13, IFN-γ and LPS as robust
Url:
DOI: 10.1038/s41598-019-44868-y
PubMed: 31171837
PubMed Central: 6554346
Affiliations:
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Satchell</name><affiliation wicri:level="1"><nlm:aff id="Aff3"><institution-wrap><institution-id institution-id-type="ISNI">0000 0004 1936 7603</institution-id><institution-id institution-id-type="GRID">grid.5337.2</institution-id><institution>Academic Renal Unit, University of Bristol,</institution></institution-wrap>Bristol, UK</nlm:aff><country xml:lang="fr">Royaume-Uni</country><wicri:regionArea>Bristol</wicri:regionArea></affiliation></author><author><name sortKey="Peak, Matthew" sort="Peak, Matthew" uniqKey="Peak M" first="Matthew" last="Peak">Matthew Peak</name><affiliation wicri:level="1"><nlm:aff id="Aff2"><institution-wrap><institution-id institution-id-type="ISNI">0000 0004 0421 1374</institution-id><institution-id institution-id-type="GRID">grid.417858.7</institution-id><institution>NIHR Alder Hey Clinical Research Facility, Alder Hey Children’s NHS Foundation Trust,</institution></institution-wrap>Liverpool, UK</nlm:aff><country xml:lang="fr">Royaume-Uni</country><wicri:regionArea>Liverpool</wicri:regionArea></affiliation></author><author><name sortKey="Beresford, Michael W" sort="Beresford, Michael W" uniqKey="Beresford M" first="Michael W." last="Beresford">Michael W. Beresford</name><affiliation wicri:level="1"><nlm:aff id="Aff1"><institution-wrap><institution-id institution-id-type="ISNI">0000 0004 1936 8470</institution-id><institution-id institution-id-type="GRID">grid.10025.36</institution-id><institution>Department of Women’s and Children’s Health, Institute of Translational Medicine,</institution><institution>University of Liverpool,</institution></institution-wrap>Liverpool, UK</nlm:aff><country xml:lang="fr">Royaume-Uni</country><wicri:regionArea>Liverpool</wicri:regionArea></affiliation><affiliation wicri:level="1"><nlm:aff id="Aff2"><institution-wrap><institution-id institution-id-type="ISNI">0000 0004 0421 1374</institution-id><institution-id institution-id-type="GRID">grid.417858.7</institution-id><institution>NIHR Alder Hey Clinical Research Facility, Alder Hey Children’s NHS Foundation Trust,</institution></institution-wrap>Liverpool, UK</nlm:aff><country xml:lang="fr">Royaume-Uni</country><wicri:regionArea>Liverpool</wicri:regionArea></affiliation><affiliation wicri:level="1"><nlm:aff id="Aff4"><institution-wrap><institution-id institution-id-type="ISNI">0000 0004 0421 1374</institution-id><institution-id institution-id-type="GRID">grid.417858.7</institution-id><institution>Department of Paediatric Rheumatology,</institution><institution>Alder Hey Children’s NHS Foundation Trust,</institution></institution-wrap>Liverpool, UK</nlm:aff><country xml:lang="fr">Royaume-Uni</country><wicri:regionArea>Liverpool</wicri:regionArea></affiliation></author></analytic><series><title level="j">Scientific Reports</title><idno type="eISSN">2045-2322</idno><imprint><date when="2019">2019</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p id="Par1">Juvenile-onset lupus nephritis (LN) affects up to 80% of juvenile-onset systemic lupus erythematosus patients (JSLE). As the exact role of human renal glomerular endothelial cells (GEnCs) in LN has not been fully elucidated, the aim of this study was to investigate their involvement in LN. Conditionally immortalised human GEnCs (ciGEnCs) were treated with pro-inflammatory cytokines known to be involved in LN pathogenesis and also with LPS. Secretion and surface expression of pro-inflammatory proteins was quantified via ELISA and flow cytometry. NF-κΒ and STAT-1 activation was investigated via immunofluorescence. Serum samples from JSLE patients and from healthy controls were used to treat ciGEnCs to determine via qRT-PCR potential changes in the mRNA levels of pro-inflammatory genes. Our results identified TNF-α, IL-1β, IL-13, IFN-γ and LPS as robust <italic>in vitro</italic> stimuli of ciGEnCs. Each of them led to significantly increased production of different pro-inflammatory proteins, including; IL-6, IL-10, MCP-1, sVCAM-1, MIP-1α, IP-10, GM-CSF, M-CSF, TNF-α, IFN-γ, VCAM-1, ICAM-1, PD-L1 and ICOS-L. TNF-α and IL-1β were shown to activate NF-κB, whilst IFN-γ activated STAT-1. JSLE patient serum promoted IL-6 and IL-1β mRNA expression. In conclusion, our <italic>in vitro</italic> model provides evidence that human GEnCs play a pivotal role in LN-associated inflammatory process.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Kamphuis, S" uniqKey="Kamphuis S">S Kamphuis</name></author><author><name sortKey="Silverman, Ed" uniqKey="Silverman E">ED Silverman</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Sprangers, B" uniqKey="Sprangers B">B Sprangers</name></author><author><name sortKey="Monahan, M" uniqKey="Monahan M">M Monahan</name></author><author><name sortKey="Appel, Gb" uniqKey="Appel G">GB Appel</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Davidson, A" uniqKey="Davidson A">A Davidson</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Lewis, Ej" uniqKey="Lewis E">EJ Lewis</name></author><author><name sortKey="Schwartz, Mm" uniqKey="Schwartz M">MM Schwartz</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Levy, Dm" uniqKey="Levy D">DM Levy</name></author><author><name sortKey="Kamphuis, S" uniqKey="Kamphuis S">S Kamphuis</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Bennett, M" uniqKey="Bennett M">M Bennett</name></author><author><name sortKey="Brunner, Hi" uniqKey="Brunner H">HI Brunner</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Cameron, Js" uniqKey="Cameron J">JS Cameron</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Lee, Bs" uniqKey="Lee B">BS Lee</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Hagelberg, S" uniqKey="Hagelberg S">S Hagelberg</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Garin, Eh" uniqKey="Garin E">EH Garin</name></author><author><name sortKey="Donnelly, Wh" uniqKey="Donnelly W">WH Donnelly</name></author><author><name sortKey="Fennell, Rs" uniqKey="Fennell R">RS Fennell</name></author><author><name sortKey="Richard, Ga" uniqKey="Richard G">GA Richard</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Yang, Ly" uniqKey="Yang L">LY Yang</name></author><author><name sortKey="Chen, Wp" uniqKey="Chen W">WP Chen</name></author><author><name sortKey="Lin, Cy" uniqKey="Lin C">CY Lin</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Baqi, N" uniqKey="Baqi N">N Baqi</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Scott, Rizaldy P" uniqKey="Scott R">Rizaldy P. 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Beresford</name><name sortKey="Beresford, Michael W" sort="Beresford, Michael W" uniqKey="Beresford M" first="Michael W." last="Beresford">Michael W. Beresford</name><name sortKey="Beresford, Michael W" sort="Beresford, Michael W" uniqKey="Beresford M" first="Michael W." last="Beresford">Michael W. Beresford</name><name sortKey="Budge, Kelly L" sort="Budge, Kelly L" uniqKey="Budge K" first="Kelly L." last="Budge">Kelly L. Budge</name><name sortKey="Midgley, Angela" sort="Midgley, Angela" uniqKey="Midgley A" first="Angela" last="Midgley">Angela Midgley</name><name sortKey="Peak, Matthew" sort="Peak, Matthew" uniqKey="Peak M" first="Matthew" last="Peak">Matthew Peak</name><name sortKey="Satchell, Simon C" sort="Satchell, Simon C" uniqKey="Satchell S" first="Simon C." last="Satchell">Simon C. Satchell</name><name sortKey="Wright, Rachael D" sort="Wright, Rachael D" uniqKey="Wright R" first="Rachael D." last="Wright">Rachael D. 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