Bioanalytical chemistry of cytokines – A review

Stenken, Julie A.; Poschenrieder, Andreas

doi:10.1016/j.aca.2014.10.009

Cited by 229 publications

(179 citation statements)

References 279 publications

(277 reference statements)

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“…However, TNF-α was almost undetectable in the conditioned medium and cell extracts from non-stimulated astrocytes, confirming that TNF-α is essentially produced by stimulated astrocytes. Since TNF-α via NF-kB activation triggers inflammatory events such as cytokines production, we also found that upon short-term TNF-α stimulation (24 h), either modulatory cytokines IL-2, IL-4 and IL-6 or the chemotactic and inflammatory cytokine IL-8 had their levels increased from pg/mL to ng/ml range, which is typical for inflammatory responses or pathological conditions (Stenken & Poschenrieder, 2015).…”

Section: Discussionmentioning

confidence: 53%

Short and long TNF-alpha exposure recapitulates canonical astrogliosis events in human induced pluripotent stem cells-derived astrocytes

Trindade

Loiola

Gasparotto

et al. 2019

Preprint

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Astrogliosis comprises a variety of changes in astrocytes that occur in a context-specific manner, triggered by temporally diverse signaling events that vary with the nature and severity of brain insults. However, most mechanisms underlying astrogliosis were described using animal models, which fail to reproduce some aspects of human astroglial signaling. Here, we report an in vitro model to study astrogliosis using human induced pluripotent stem cells (iPSC)-derived astrocytes which replicates aspects temporally intertwined of reactive astrocytes in vivo. We analyzed the time course of astrogliosis by measuring nuclear translocation of NF-kB, secretion of cytokines and changes in morphological phenotypes of human iPSC-derived astrocytes exposed to TNF-α. It was observed the NF-kB nuclear translocation, increases either in the inflammation-related cytokines secretion and gene expression for IL-1β, IL-6 and TNF-α following 24 h TNF-α stimulation. After 5 days, human iPSCderived astrocytes exposed to TNF-α exhibited increases in vimentin and GFAP immunolabeling, elongated shape and shrinkage of nuclei, which is typical phenotypes of astrogliosis. Moreover, about a 50% decrease in D-[ 3 H] aspartate uptake was observed over the astrogliosis course with no evident cell damage, which suggests astrocytic dysfunction. Taken together, our results indicate that cultured human iPSC-derived astrocytes reproduce canonical events associated to astrogliosis in a time dependent fashion. Our findings may contribute to a better understanding of mechanisms governing human astrogliosis. Furthermore, the approach described here presents a potential applicability as a platform to uncover novel biomarkers and new drug targets to refrain astrogliosis associated to human brain disorders.

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Section: Discussionmentioning

confidence: 53%

Short and long TNF-alpha exposure recapitulates canonical astrogliosis events in human induced pluripotent stem cells-derived astrocytes

Trindade

Loiola

Gasparotto

et al. 2019

Preprint

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“…In order to make such measurements valuable for reliable assessments, the analytical requirements are very high. Typically, the most effective cytokines work at very low concentration ranges [29] and have to be determined out of complex biological matrices such as blood plasma consisting of thousands of different molecules [30]. Furthermore, it may be desirable to determine several proteins occurring at quite different concentration ranges within the same sample.…”

Section: Discussionmentioning

confidence: 99%

Quantification of cytokines secreted by primary human cells using multiple reaction monitoring: evaluation of analytical parameters

et al. 2015

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Determination of secreted proteins provides highly valuable information about cell functions. While the typical methods for the determination of biologically relevant but low abundant molecular species still rely on the use of specific antibodies, mass spectrometry-based methods are now gaining sufficient sensitivity to cope with such challenges as well. In the current study, we have identified several cytokines and chemokines which were induced in primary human umbilical vein endothelial cells upon inflammatory activation. Based on the high-resolution mass spectrometry data obtained with a Q Exactive orbitrap, we built an MRM method to quantify the most relevant molecules selected from the screening experiment. All experimental data are available via ProteomeXchange, PXD002211/12, and Panorama ( www.panoramaweb.org ). Using nano-flow Chip-HPLC coupled to a 6490 triple-quadrupole MS for MRM analyses, we achieved calibration curves covering a linear range of four orders of magnitude and detection limits in the low attomol per microliter concentration range. Carryover was consistently less than 0.005 %, the accuracy was between 80 and 120 %, and the median coefficient of variation for LC/MS was only 2.2 %. When including the variance of quantification introduced by cell culture and digestion, the coefficient of variation was less than 20 % for most peptides. With appropriate marker molecules, we monitored typical variations introduced by cell culture caused by differences in cell numbers, proliferative states, and cell death. As a result, here, we present a robust and efficient MRM-based assay for the accurate and sensitive determination of cytokines and chemokines representative for functional cell states and including comprehensive quality controls. Graphical Abstract Work flow diagram: Data processing steps beginning with orbitrap-based shotgun data acquisition and MaxQuant data analysis, followed by peptide and transition selection for MRM analysis using Skyline and experimental validation using triple quadrupole MS.

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“…Cytokines are low molecular weight bioactive proteins produced by many different cells and strongly associated with the immune system (Stenken and Poschenrieder, 2015) . There is enormous clinical interest in cytokines determination as elevated concentrations of these proteins are associated with inflammation or disease progression and, therefore, various types of cytokines are widely used as biomarkers to characterize the immune function, predict diseases and monitor their evolution and treatments.…”

Section: Introductionmentioning

confidence: 99%

Electrochemical immunosensor for sensitive determination of transforming growth factor (TGF) - β1 in urine

Sánchez‐Tirado

Pérez‐Quintanilla

González‐Cortés

et al. 2017

Biosensors and Bioelectronics

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The first amperometric immunosensor for the quantification of TGF-β1, a cytokine proposed as a biomarker for patients having or at risk for renal disease, is described in this work. The immunosensor design involves disposable devices using carboxylic acid-functionalized magnetic microparticles supported onto screen-printed carbon electrodes and covalent immobilization of the specific antibody for TGF-β1 using Mix&Go polymer. A sandwich-type immunoassay was performed using biotin-anti-TGF and conjugation with peroxidase-labeled streptavidin (poly-HRP-Strept) polymer. Amperometric measurements were carried out at -0.20V by adding hydrogen peroxide solution onto the electrode surface in the presence of hydroquinone as the redox mediator. The calibration plot allowed a range of linearity extending between 15 and 3000pg/mL TGF-β1 which is adequate for the determination of the cytokine in plasma and urine. The limit of detection, 10pg/mL, is notably improved with respect to those obtained with ELISA kits. The usefulness of the immunosensor for the determination of low TGF-β1 concentrations in real samples was evaluated by analyzing spiked urine at different pg/mL concentration levels.

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Bioanalytical chemistry of cytokines – A review

Cited by 229 publications

References 279 publications

Short and long TNF-alpha exposure recapitulates canonical astrogliosis events in human induced pluripotent stem cells-derived astrocytes

Short and long TNF-alpha exposure recapitulates canonical astrogliosis events in human induced pluripotent stem cells-derived astrocytes

Quantification of cytokines secreted by primary human cells using multiple reaction monitoring: evaluation of analytical parameters

Electrochemical immunosensor for sensitive determination of transforming growth factor (TGF) - β1 in urine

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