Studies on Huntington's disease (HD) demonstrated altered immune response in HD gene carriers. Using multiplexing immunoassay, we simultaneously investigated seven cytokines in secretomes of microglia and blood monocytes, cerebrospinal fluid (CSF) and serum collected from transgenic HD minipigs at pre-symptomatic disease stage. Decline in IFNα and IL-10 was observed in CSF and secretome of microglia whilst elevated IL-8 and IL-1β levels were secreted by microglia. Additionally, IL-8 was increased in serum. The proportion of mutant huntingtin in microglia may have causative impact on cytokine production. IFNα, IL-10, IL-8 and IL-1β represent promising biomarkers reflecting immuno-pathological mechanisms in porcine HD model.
- MeSH
- biologické markery metabolismus MeSH
- centrální nervový systém cytologie patologie MeSH
- cytokiny metabolismus MeSH
- DNA vazebné proteiny metabolismus MeSH
- geneticky modifikovaná zvířata MeSH
- Huntingtonova nemoc genetika metabolismus patologie MeSH
- interferon alfa metabolismus MeSH
- interleukin-10 metabolismus MeSH
- interleukin-1beta metabolismus MeSH
- interleukin-8 metabolismus MeSH
- kultivované buňky MeSH
- lidé MeSH
- lipopolysacharidy farmakologie MeSH
- mikroglie účinky léků metabolismus MeSH
- miniaturní prasata MeSH
- modely nemocí na zvířatech MeSH
- monocyty účinky léků metabolismus MeSH
- prasata MeSH
- proteiny nervové tkáně genetika metabolismus MeSH
- regulace genové exprese genetika MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Immunoassays represent valuable and broadly used techniques for detection and quantification of proteins. Thanks to their high sensitivity, such techniques are powerful for analyzing growth factors, trophic factors, angiogenic factors, hormones, cytokines, chemokines, soluble receptors, and other proteins which play key roles in intercellular communication and operate as potent regulators of stem cell survival, proliferation, differentiation, or cell death. Multiplex immunological assays, in contrast to ELISA, offer simultaneous quantification of tens of proteins across multiple samples, and have been developed to save time, costs, and sample volumes. Among them, planar antibody microarrays and xMAP(®) bead-based assays have become particularly popular for characterization of proteins secreted by stem cells, as they are relatively easy, highly accurate, multiplex to a high degree and a broad spectrum of analytes can be measured. Here, we describe protocols for multiplex quantification of secreted proteins using Quantibody(®) microarrays (RayBiotech) and xMAP(®) assays (Luminex and its partners).
- MeSH
- buněčné kultury MeSH
- čipová analýza proteinů metody MeSH
- cytokiny metabolismus MeSH
- imunoanalýza metody MeSH
- kmenové buňky fyziologie MeSH
- kultivační média speciální metabolismus MeSH
- kultivované buňky MeSH
- mezibuněčná komunikace * MeSH
- mezibuněčné signální peptidy a proteiny metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
