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Conference Paper: Anti-dsDNA antibodies bind to Ku70 in proximal renal tubular epithelial cells and increased matrix protein and cytokine secretion

TitleAnti-dsDNA antibodies bind to Ku70 in proximal renal tubular epithelial cells and increased matrix protein and cytokine secretion
Authors
Issue Date2018
PublisherAmerican Society of Nephrology. The Journal's web site is located at http://www.jasn.org
Citation
American Society of Nephrology (ASN) Kidney Week 2018, San Diego, California, USA, 23-28 October 2018. In Journal of the American Society of Nephrology, 2018, v. 29 n. Abstract Suppl., p. 5 How to Cite?
AbstractBackground: Anti-dsDNA antibodies can lead to immune-mediated kidney injury through complement activation or trigger downstream pro-inflammatory or fibrotic responses after cellular binding. Immune deposition along tubular basement membrane is commonly observed. We investigated the binding of anti-dsDNA antibodies to proximal renal tubular epithelial cells (PTEC). Methods: Human polyclonal anti-dsDNA antibodies were isolated from the sera of lupus nephritis patients using affinity chromatography and samples demonstrating high binding affinity to PTEC were studied. Cultured PTEC were incubated with serum-free medium, control IgG, or anti-dsDNA antibodies for up to 48h. PTEC plasma membrane proteins were isolated and immuno-precipitated with anti-dsDNA antibodies to identify cross-reactive antigens using LC-MS/MS. The role of Ku70 in inflammatory and fibrotic processes was investigated by gene silencing with RNAi. Results: The 70 kDa on PTEC cell membrane that bound anti-dsDNA antibodies was identified as Ku70 by LC-MS/MS. Exogenous DNA, histones, or nucleosomes did not affect its binding by anti-dsDNA antibodies. Anti-dsDNA antibodies binding to Ku70 was accompanied by increased fibronectin and laminin expression, and increased MCP-1 and IL-6 secretion (P<0.05, for all). Ku70 gene silencing with ninety percent efficacy resulted in reduced PTEC binding by anti-dsDNA antibodies as shown by immunohistochemistry. Ku70 knockdown significantly decreased the expression of fibronectin, laminin, and Bax, and and also the secretion of MCP-1 and IL-6, induced by TGF-β1, MCP-1, IL-1β, and IL-6 (P<0.05, for all). Kidney specimens from lupus nephritis patients and NZB/W F1 mice showed markedly increased Ku70 expression in the proximal tubules Conclusion: Our data shows that Ku70 mediates the binding of anti-dsDNA antibodies to PTEC, which is accompanied by downstream pro-inflammatory and pro-fibrotic cellular responses.
DescriptionOral Presentation - Session: CKD: Mechanisms of Kidney Fibrosis - abstract no. TH-OR020
Persistent Identifierhttp://hdl.handle.net/10722/273060
ISSN
2019 Impact Factor: 9.274
2015 SCImago Journal Rankings: 4.699

 

DC FieldValueLanguage
dc.contributor.authorChan, DTM-
dc.contributor.authorHo, SSK-
dc.contributor.authorTai, CP-
dc.contributor.authorChan, CYC-
dc.contributor.authorCheng, WM-
dc.contributor.authorChau, KM-
dc.contributor.authorYung, SSY-
dc.date.accessioned2019-08-06T09:21:48Z-
dc.date.available2019-08-06T09:21:48Z-
dc.date.issued2018-
dc.identifier.citationAmerican Society of Nephrology (ASN) Kidney Week 2018, San Diego, California, USA, 23-28 October 2018. In Journal of the American Society of Nephrology, 2018, v. 29 n. Abstract Suppl., p. 5-
dc.identifier.issn1046-6673-
dc.identifier.urihttp://hdl.handle.net/10722/273060-
dc.descriptionOral Presentation - Session: CKD: Mechanisms of Kidney Fibrosis - abstract no. TH-OR020-
dc.description.abstractBackground: Anti-dsDNA antibodies can lead to immune-mediated kidney injury through complement activation or trigger downstream pro-inflammatory or fibrotic responses after cellular binding. Immune deposition along tubular basement membrane is commonly observed. We investigated the binding of anti-dsDNA antibodies to proximal renal tubular epithelial cells (PTEC). Methods: Human polyclonal anti-dsDNA antibodies were isolated from the sera of lupus nephritis patients using affinity chromatography and samples demonstrating high binding affinity to PTEC were studied. Cultured PTEC were incubated with serum-free medium, control IgG, or anti-dsDNA antibodies for up to 48h. PTEC plasma membrane proteins were isolated and immuno-precipitated with anti-dsDNA antibodies to identify cross-reactive antigens using LC-MS/MS. The role of Ku70 in inflammatory and fibrotic processes was investigated by gene silencing with RNAi. Results: The 70 kDa on PTEC cell membrane that bound anti-dsDNA antibodies was identified as Ku70 by LC-MS/MS. Exogenous DNA, histones, or nucleosomes did not affect its binding by anti-dsDNA antibodies. Anti-dsDNA antibodies binding to Ku70 was accompanied by increased fibronectin and laminin expression, and increased MCP-1 and IL-6 secretion (P<0.05, for all). Ku70 gene silencing with ninety percent efficacy resulted in reduced PTEC binding by anti-dsDNA antibodies as shown by immunohistochemistry. Ku70 knockdown significantly decreased the expression of fibronectin, laminin, and Bax, and and also the secretion of MCP-1 and IL-6, induced by TGF-β1, MCP-1, IL-1β, and IL-6 (P<0.05, for all). Kidney specimens from lupus nephritis patients and NZB/W F1 mice showed markedly increased Ku70 expression in the proximal tubules Conclusion: Our data shows that Ku70 mediates the binding of anti-dsDNA antibodies to PTEC, which is accompanied by downstream pro-inflammatory and pro-fibrotic cellular responses.-
dc.languageeng-
dc.publisherAmerican Society of Nephrology. The Journal's web site is located at http://www.jasn.org-
dc.relation.ispartofJournal of the American Society of Nephrology-
dc.relation.ispartofAmerican Society of Nephrology Kidney Week 2018-
dc.titleAnti-dsDNA antibodies bind to Ku70 in proximal renal tubular epithelial cells and increased matrix protein and cytokine secretion-
dc.typeConference_Paper-
dc.identifier.emailChan, DTM: dtmchan@hkucc.hku.hk-
dc.identifier.emailTai, CP: cpandrew@hku.hk-
dc.identifier.emailChan, CYC: calebccy@hku.hk-
dc.identifier.emailYung, SSY: ssyyung@hku.hk-
dc.identifier.authorityChan, DTM=rp00394-
dc.identifier.authorityYung, SSY=rp00455-
dc.identifier.hkuros299784-
dc.identifier.volume29-
dc.identifier.issueAbstract Suppl.-
dc.identifier.spage5-
dc.identifier.epage5-
dc.publisher.placeUnited States-

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