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Conference Paper: Anti-dsDNA antibodies from patients with lupus nephritis bind to annexin II on human mesangial cells

TitleAnti-dsDNA antibodies from patients with lupus nephritis bind to annexin II on human mesangial cells
Authors
Issue Date2010
PublisherAmerican Society of Nephrology. The Journal's web site is located at http://www.jasn.org
Citation
Renal Week 2010 - The 43rd Annual Meeting of the American Society of Nephrology, Denver, CO., 16-21 November 2010. In Journal of the American Society of Nephrology, 2010 meeting abstracts, p. 755A, SA-PO2815 How to Cite?
AbstractLupus nephritis is characterized by the production of anti-dsDNA antibodies and proliferative glomerulonephritis. The mechanism through which anti-dsDNA antibodies interact with resident renal cells remains to be elucidated. We and others have previously demonstrated that human anti-dsDNA antibodies can bind to human mesangial cells (HMC) without the need of bridging chromatin material. In this study, we characterized the cross-reactive antigen(s) on HMC that mediate the binding. Human polyclonal anti-dsDNA antibodies were isolated from patients with lupus nephritis using Protein A-Sepharose followed by DNA-cellulose affinity chromatography. The binding of anti-dsDNA antibodies to HMC was assessed by flow cytometry and cellular ELISA. HMC plasma membrane fractions were subjected to Western blotting, probed with purified anti-dsDNA antibodies, and analyzed with MALDI-TOF spectrometry to identify ‘cross-reactive’ membrane proteins. Renal biopsies were examined with immunohistochemistry. Limited trypsin but not DNase treatment of HMC significantly reduced anti-dsDNA antibody binding (P<0.05). Anti-dsDNA antibodies predominantly bound to a cell surface antigen with molecular weight of ∼36kDa. This band was identified as annexin II by MALDI-TOF spectrometry. The binding activity between anti-dsDNA antibodies and annexin II correlated with disease activity. Glomerular annexin II expression was significantly increased in active lupus nephritis compared with controls and non-lupus kidney diseases (P<0.05), and co-localized with IgG and C3 deposition. Our data demonstrate that annexin II on the plasma membrane of HMC mediates anti-dsDNA antibody binding.
DescriptionImmunology, Pathology: Basic/Experimental Immunology I (Poster): SA-PO2815
Abstract Supplement of JASN is located at http://www.asn-online.org/education/kidneyweek/archives/
Persistent Identifierhttp://hdl.handle.net/10722/135921
ISSN
2015 Impact Factor: 8.491
2015 SCImago Journal Rankings: 4.699

 

DC FieldValueLanguage
dc.contributor.authorYung, Sen_US
dc.contributor.authorCheung, KFen_US
dc.contributor.authorZhang, Qen_US
dc.contributor.authorChan, DTMen_US
dc.date.accessioned2011-07-27T01:59:43Z-
dc.date.available2011-07-27T01:59:43Z-
dc.date.issued2010en_US
dc.identifier.citationRenal Week 2010 - The 43rd Annual Meeting of the American Society of Nephrology, Denver, CO., 16-21 November 2010. In Journal of the American Society of Nephrology, 2010 meeting abstracts, p. 755A, SA-PO2815en_US
dc.identifier.issn1046-6673-
dc.identifier.urihttp://hdl.handle.net/10722/135921-
dc.descriptionImmunology, Pathology: Basic/Experimental Immunology I (Poster): SA-PO2815-
dc.descriptionAbstract Supplement of JASN is located at http://www.asn-online.org/education/kidneyweek/archives/-
dc.description.abstractLupus nephritis is characterized by the production of anti-dsDNA antibodies and proliferative glomerulonephritis. The mechanism through which anti-dsDNA antibodies interact with resident renal cells remains to be elucidated. We and others have previously demonstrated that human anti-dsDNA antibodies can bind to human mesangial cells (HMC) without the need of bridging chromatin material. In this study, we characterized the cross-reactive antigen(s) on HMC that mediate the binding. Human polyclonal anti-dsDNA antibodies were isolated from patients with lupus nephritis using Protein A-Sepharose followed by DNA-cellulose affinity chromatography. The binding of anti-dsDNA antibodies to HMC was assessed by flow cytometry and cellular ELISA. HMC plasma membrane fractions were subjected to Western blotting, probed with purified anti-dsDNA antibodies, and analyzed with MALDI-TOF spectrometry to identify ‘cross-reactive’ membrane proteins. Renal biopsies were examined with immunohistochemistry. Limited trypsin but not DNase treatment of HMC significantly reduced anti-dsDNA antibody binding (P<0.05). Anti-dsDNA antibodies predominantly bound to a cell surface antigen with molecular weight of ∼36kDa. This band was identified as annexin II by MALDI-TOF spectrometry. The binding activity between anti-dsDNA antibodies and annexin II correlated with disease activity. Glomerular annexin II expression was significantly increased in active lupus nephritis compared with controls and non-lupus kidney diseases (P<0.05), and co-localized with IgG and C3 deposition. Our data demonstrate that annexin II on the plasma membrane of HMC mediates anti-dsDNA antibody binding.-
dc.languageengen_US
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 Nephrologyen_US
dc.titleAnti-dsDNA antibodies from patients with lupus nephritis bind to annexin II on human mesangial cellsen_US
dc.typeConference_Paperen_US
dc.identifier.emailYung, S: ssyyung@hku.hken_US
dc.identifier.emailCheung, KF: skfc819@hku.hken_US
dc.identifier.emailZhang, Q: zhjhr@hkucc.hku.hken_US
dc.identifier.emailChan, DTM: dtmchan@hku.hk-
dc.identifier.authorityYung, S=rp00455en_US
dc.identifier.authorityChan, DTM=rp00394en_US
dc.description.naturelink_to_OA_fulltext-
dc.identifier.hkuros188593en_US
dc.identifier.volume2010en_US
dc.identifier.issuemeeting abstracts-
dc.identifier.spage755Aen_US
dc.identifier.epage755Aen_US
dc.publisher.placeUnited States-

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