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Article: Synthesis of TNF-α by mesangial cells cultured with polymeric anionic IgA - Role of MAPK and NF-κB

TitleSynthesis of TNF-α by mesangial cells cultured with polymeric anionic IgA - Role of MAPK and NF-κB
Authors
KeywordsAnionic IgA
Human mesangial cell
IgA nephropathy
Polymeric IgA
Issue Date2008
PublisherOxford University Press. The Journal's web site is located at http://ndt.oxfordjournals.org/
Citation
Nephrology Dialysis Transplantation, 2008, v. 23 n. 1, p. 72-81 How to Cite?
AbstractBackground. Deposition of polymeric IgA1 (pIgA) in kidney mesangium is the hallmark of IgA nephropathy (IgAN). Current consensus is that a fraction of IgA1 molecules in the circulation of IgAN patients exhibit aberrant structures or properties that may lead to their deposition. Our previous findings suggest that the anionic property of IgA1 may play a role in mesangial IgA1 deposition in patients with IgAN. In the present study, the functional consequences of the binding of anionic polymeric IgA1 to human mesangial cells (HMCs) were investigated. Methods. Anionic polymeric IgA1 from IgAN patients and healthy subjects was isolated by sequential jacalin affinity chromatography, size exclusion chromatography using size exclusion and MonoQ ion exchange chromatography. HMCs were cultured with purified anionic polymeric IgA1 and the release of tumour necrosis factor-α (TNF-α) and interleukin-6 (IL-6) was examined by enzyme-linked immunosorbent assay. The signalling pathways involved in anionic pIgA-mediated HMC activation were examined by immunoblotting. Standard electrophoretic mobility shift assay (EMSA) was used to further examine whether the transcriptional factor NF-κB is associated in the signalling process. To define the mechanism of TNF-α and IL-6 production, HMCs were cultured with anionic pIgA in the presence or absence of p42/p44 mitogen-activated protein kinase (MAPK) inhibitor (PD98059), NF-κB inhibitor pyrrolidine dithiocarbamate (PDTC) or NF-κB blocking permeable peptides. Results. Compared with less anionic pIgA or monomeric IgA1 (mIgA), anionic pIgA from patient with IgAN significantly increased cell proliferation (P < 0.05) when cultured with HMC. These anionic pIgA significantly increased the synthesis of TNF-α (P < 0.05) and IL-6 (P < 0.05) in a dose and time-dependent manner. Furthermore, the increased synthesis of IL-6 and TNF-α by anionic pIgA in HMC was significantly diminished (P < 0.01) in the presence of NF-κB inhibitor pyrrolidine dithiocarbamate and NF-κB blocking permeable peptides SN50 (P < 0.01). The increased synthesis of IL-6 by anionic pIgA in HMC was reduced by inhibitor to NF-κB or p42/p44 MAPK and was abolished by the simultaneous presence of inhibitors to p42/p44 MAPK and NF-κB. The up-regulation of TNF-α was partially suppressed by inhibitor to NF-κB but not PD98059. Conclusion. Our results suggest that polymeric anionic IgA1 could activate HMC and increase the synthesis of TNF-α and IL-6. While both the p42/p44 MAPK and NF-κB pathways are essential in regulating the anionic pIgA-induced synthesis of IL-6, TNF-α synthesis mediated by anionic pIgA is partly dependent on NF-κB. © The Author [2007]. Published by Oxford University Press on behalf of ERA-EDTA. All rights reserved.
Persistent Identifierhttp://hdl.handle.net/10722/163172
ISSN
2015 Impact Factor: 4.085
2015 SCImago Journal Rankings: 1.780
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorLeung, JCKen_HK
dc.contributor.authorTang, SCWen_HK
dc.contributor.authorChan, LYYen_HK
dc.contributor.authorChan, WLen_HK
dc.contributor.authorLai, KNen_HK
dc.date.accessioned2012-09-05T05:28:25Z-
dc.date.available2012-09-05T05:28:25Z-
dc.date.issued2008en_HK
dc.identifier.citationNephrology Dialysis Transplantation, 2008, v. 23 n. 1, p. 72-81en_HK
dc.identifier.issn0931-0509en_HK
dc.identifier.urihttp://hdl.handle.net/10722/163172-
dc.description.abstractBackground. Deposition of polymeric IgA1 (pIgA) in kidney mesangium is the hallmark of IgA nephropathy (IgAN). Current consensus is that a fraction of IgA1 molecules in the circulation of IgAN patients exhibit aberrant structures or properties that may lead to their deposition. Our previous findings suggest that the anionic property of IgA1 may play a role in mesangial IgA1 deposition in patients with IgAN. In the present study, the functional consequences of the binding of anionic polymeric IgA1 to human mesangial cells (HMCs) were investigated. Methods. Anionic polymeric IgA1 from IgAN patients and healthy subjects was isolated by sequential jacalin affinity chromatography, size exclusion chromatography using size exclusion and MonoQ ion exchange chromatography. HMCs were cultured with purified anionic polymeric IgA1 and the release of tumour necrosis factor-α (TNF-α) and interleukin-6 (IL-6) was examined by enzyme-linked immunosorbent assay. The signalling pathways involved in anionic pIgA-mediated HMC activation were examined by immunoblotting. Standard electrophoretic mobility shift assay (EMSA) was used to further examine whether the transcriptional factor NF-κB is associated in the signalling process. To define the mechanism of TNF-α and IL-6 production, HMCs were cultured with anionic pIgA in the presence or absence of p42/p44 mitogen-activated protein kinase (MAPK) inhibitor (PD98059), NF-κB inhibitor pyrrolidine dithiocarbamate (PDTC) or NF-κB blocking permeable peptides. Results. Compared with less anionic pIgA or monomeric IgA1 (mIgA), anionic pIgA from patient with IgAN significantly increased cell proliferation (P < 0.05) when cultured with HMC. These anionic pIgA significantly increased the synthesis of TNF-α (P < 0.05) and IL-6 (P < 0.05) in a dose and time-dependent manner. Furthermore, the increased synthesis of IL-6 and TNF-α by anionic pIgA in HMC was significantly diminished (P < 0.01) in the presence of NF-κB inhibitor pyrrolidine dithiocarbamate and NF-κB blocking permeable peptides SN50 (P < 0.01). The increased synthesis of IL-6 by anionic pIgA in HMC was reduced by inhibitor to NF-κB or p42/p44 MAPK and was abolished by the simultaneous presence of inhibitors to p42/p44 MAPK and NF-κB. The up-regulation of TNF-α was partially suppressed by inhibitor to NF-κB but not PD98059. Conclusion. Our results suggest that polymeric anionic IgA1 could activate HMC and increase the synthesis of TNF-α and IL-6. While both the p42/p44 MAPK and NF-κB pathways are essential in regulating the anionic pIgA-induced synthesis of IL-6, TNF-α synthesis mediated by anionic pIgA is partly dependent on NF-κB. © The Author [2007]. Published by Oxford University Press on behalf of ERA-EDTA. All rights reserved.en_HK
dc.languageengen_US
dc.publisherOxford University Press. The Journal's web site is located at http://ndt.oxfordjournals.org/en_HK
dc.relation.ispartofNephrology Dialysis Transplantationen_HK
dc.rightsNephrology, Dialysis, Transplantation. Copyright © Oxford University Press.-
dc.subjectAnionic IgAen_HK
dc.subjectHuman mesangial cellen_HK
dc.subjectIgA nephropathyen_HK
dc.subjectPolymeric IgAen_HK
dc.titleSynthesis of TNF-α by mesangial cells cultured with polymeric anionic IgA - Role of MAPK and NF-κBen_HK
dc.typeArticleen_HK
dc.identifier.emailLeung, JCK: jckleung@hku.hken_HK
dc.identifier.emailTang, SCW: scwtang@hku.hken_HK
dc.identifier.emailLai, KN: knlai@hku.hken_HK
dc.identifier.authorityLeung, JCK=rp00448en_HK
dc.identifier.authorityTang, SCW=rp00480en_HK
dc.identifier.authorityLai, KN=rp00324en_HK
dc.description.naturelink_to_OA_fulltexten_US
dc.identifier.doi10.1093/ndt/gfm581en_HK
dc.identifier.pmid17898021-
dc.identifier.scopuseid_2-s2.0-44449168619en_HK
dc.identifier.hkuros143117-
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-44449168619&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume23en_HK
dc.identifier.issue1en_HK
dc.identifier.spage72en_HK
dc.identifier.epage81en_HK
dc.identifier.isiWOS:000253022100017-
dc.publisher.placeUnited Kingdomen_HK
dc.identifier.scopusauthoridLeung, JCK=7202180349en_HK
dc.identifier.scopusauthoridTang, SCW=7403437082en_HK
dc.identifier.scopusauthoridChan, LYY=55182644100en_HK
dc.identifier.scopusauthoridChan, WL=24333977900en_HK
dc.identifier.scopusauthoridLai, KN=7402135706en_HK

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