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Quantitative mass spectrometry to study inflammatory cartilage degradation and resulting interactions with the complement system

Furst, Camilla Melin LU ; Åhrman, Emma LU ; Bratteby, Klas; Waldemarson, Sofia LU ; Malmström, Johan LU and Blom, Anna M. LU (2016) In Journal of Immunology 197(8). p.3415-3424
Abstract

Joint diseases are often characterized by inflammatory processes that result in pathological changes in joint tissues, including cartilage degradation and release of components into the synovial fluid. The complement system plays a central role in promoting the inflammation. Because several cartilage proteins are known to interact with complement, causing either activation or inhibition of the system, we aimed to investigate these interactions comprehensively. Bovine cartilage explants were cultured with IL-1α to induce cartilage degradation, followed by incubation with human serum. Label-free selected reaction monitoring mass spectrometry was used to specifically quantify complement proteins interacting with the cartilage explant. In... (More)

Joint diseases are often characterized by inflammatory processes that result in pathological changes in joint tissues, including cartilage degradation and release of components into the synovial fluid. The complement system plays a central role in promoting the inflammation. Because several cartilage proteins are known to interact with complement, causing either activation or inhibition of the system, we aimed to investigate these interactions comprehensively. Bovine cartilage explants were cultured with IL-1α to induce cartilage degradation, followed by incubation with human serum. Label-free selected reaction monitoring mass spectrometry was used to specifically quantify complement proteins interacting with the cartilage explant. In parallel, the time-dependent degradation of cartilage was detected using mass spectrometry analysis (liquid chromatography-tandem mass spectrometry). Complement proteins resulting from activation of the classical, alternative, and terminal pathways were detected on IL-1α-stimulated cartilage at time points when clear alterations in extracellular matrix composition had occurred. Increased levels of the complement activation product C4d, as detected by ELISA in serum after incubation with IL-1α-stimulated cartilage, confirmed the selected reaction monitoring results indicating complement activation. Further, typical activated (cleaved) C3 fragments were detected by Western blotting in extracts of IL-1α-stimulated cartilage. No complement activation was triggered by cartilage cultured in the absence of IL-1α. Components released from IL-1α-stimulated cartilage during culture had an inhibitory effect on complement activation. These were released after a longer incubation period with IL-1α and may represent a feedback reaction to cartilage-triggered complement activation observed after a shorter incubation period.

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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Journal of Immunology
volume
197
issue
8
pages
10 pages
publisher
American Association of Immunologists
external identifiers
  • scopus:84991491214
  • wos:000387965100042
ISSN
0022-1767
DOI
10.4049/jimmunol.1601006
language
English
LU publication?
yes
id
43bd8191-ed16-46d0-9f6e-b730b13a15a2
date added to LUP
2016-10-31 09:19:29
date last changed
2017-09-03 05:14:21
@article{43bd8191-ed16-46d0-9f6e-b730b13a15a2,
  abstract     = {<p>Joint diseases are often characterized by inflammatory processes that result in pathological changes in joint tissues, including cartilage degradation and release of components into the synovial fluid. The complement system plays a central role in promoting the inflammation. Because several cartilage proteins are known to interact with complement, causing either activation or inhibition of the system, we aimed to investigate these interactions comprehensively. Bovine cartilage explants were cultured with IL-1α to induce cartilage degradation, followed by incubation with human serum. Label-free selected reaction monitoring mass spectrometry was used to specifically quantify complement proteins interacting with the cartilage explant. In parallel, the time-dependent degradation of cartilage was detected using mass spectrometry analysis (liquid chromatography-tandem mass spectrometry). Complement proteins resulting from activation of the classical, alternative, and terminal pathways were detected on IL-1α-stimulated cartilage at time points when clear alterations in extracellular matrix composition had occurred. Increased levels of the complement activation product C4d, as detected by ELISA in serum after incubation with IL-1α-stimulated cartilage, confirmed the selected reaction monitoring results indicating complement activation. Further, typical activated (cleaved) C3 fragments were detected by Western blotting in extracts of IL-1α-stimulated cartilage. No complement activation was triggered by cartilage cultured in the absence of IL-1α. Components released from IL-1α-stimulated cartilage during culture had an inhibitory effect on complement activation. These were released after a longer incubation period with IL-1α and may represent a feedback reaction to cartilage-triggered complement activation observed after a shorter incubation period.</p>},
  author       = {Furst, Camilla Melin and Åhrman, Emma and Bratteby, Klas and Waldemarson, Sofia and Malmström, Johan and Blom, Anna M.},
  issn         = {0022-1767},
  language     = {eng},
  month        = {10},
  number       = {8},
  pages        = {3415--3424},
  publisher    = {American Association of Immunologists},
  series       = {Journal of Immunology},
  title        = {Quantitative mass spectrometry to study inflammatory cartilage degradation and resulting interactions with the complement system},
  url          = {http://dx.doi.org/10.4049/jimmunol.1601006},
  volume       = {197},
  year         = {2016},
}