Novel method for the measurement of cytokine production by a one-stage procedure

Groote, Donat De; Gevaert, Y.; López, Miguel Fraile; Gathy, R.; Fauchet, F.; Dehart, I.; Jadoul, Michel; Radoux, D.; Franchimont, P

doi:10.1016/0022-1759(93)90130-y

Cited by 57 publications

(23 citation statements)

References 16 publications

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“…Methods for separating peripheral blood mononuclear cells (PBMC), such as Ficoll-Hypaque separation, have previously been found to alter the composition of T-cell subsets and decrease cell proliferation and cytokine production [23][24][25].…”

Section: Discussionmentioning

confidence: 99%

Stimulation of whole blood cultures in patients with ankylosing spondylitis by a mitogen derived fromMycoplasma arthritidis (MAS) and other mitogens

et al. 1997

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In this study we compared cytokine production and cell proliferation of immunocompetent cells derived from patients with ankylosing spondylitis (AS) to those from healthy blood donors using a whole blood assay. To this end, blood cell cultures were stimulated with the superantigens MAS (Mycoplasma arthritidis supernatant) and staphylococcal enterotoxin B (SEB) and the plant lectins phytohaemagglutinin (PHA) and concanavalin A (Con A). The number of white blood cells (WBC) and lymphocyte subsets were also determined. Cell proliferation and levels of interferon-gamma (IFN-gamma), interleukin-1 beta (IL-1 beta) and interleukin-6 (IL-6) were measured after stimulation with the different mitogens. An ELISA test was used to analyse supernatant cytokine levels. Individuals with AS showed significantly lower IFN-gamma concentrations and markedly lower cell proliferation rates with all tested mitogens than healthy controls, while there was no significant difference in IL-6 synthesis. IL-1 beta levels were slightly impaired in the patient group, but only blood cell cultures stimulates with MAS showed a statistical significance. Furthermore, there was a significant elevation of leucocytes and lymphocytes in patients with AS resulting in higher numbers of CD4-positive cells, which implies a higher CD4:CD8 cell ratio. CD19- and CD8-positive cells were not significantly distinct compared to healthy controls. This deviation in cytokine levels and cell proliferation points to a suppression of T lymphocytes. A disturbed T-lymphocyte function may play a part in the pathogenesis of AS.

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Section: Discussionmentioning

confidence: 99%

Stimulation of whole blood cultures in patients with ankylosing spondylitis by a mitogen derived fromMycoplasma arthritidis (MAS) and other mitogens

et al. 1997

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“…The same authors, found that there was a trend toward a significant decreased secretion of IFNy by stimulated T lymphocytes preincubated with the above antidepressants. Since diluted whole blood stimulated with phytohemagglutinin (PHA) plus lipopolysaccharide (LPS) offers the most appropriate and reproducible culture condition in order to measure cytokines, such as IFNy, IL-6, and IL-1 (reviews: De Groote, Zangerle, Gevaert, Fassotte, Beguin, Noizat-Pirenne, Pirenne, Gathy, Lopez, Dehart, Igot, Baudrihaye, Delacroix, & Franchimont, 1992;De Groote, Gevaert, Lopez, Gathy, Fauchet, Dehart, Jadoul, Radoux, & Franchimont, 1993), our laboratory has examined the effects of antidepressive agents on cytokine production by diluted whole blood cultures. The focus of interest was the IFNy/IL-10 ratio, since it is of critical importance in determining the capacity of supernatants to activate or inhibit monocytic and T lymphocytic functions (Katsikis et al, 1995).…”

Section: In Vitro Effects Of Antidepressantsmentioning

confidence: 99%

Major Depression and Activation of The Inflammatory Response System

Maes¹

1999

Advances in Experimental Medicine and Biology

539

345

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“…The most promising approaches for the development of CsA bioassays are based on whole blood stimulation and the subsequent measurement of direct or indirect targets of CsA action in T cells. Among them are assays determining calcineurin activity (27,28), cytokine, in particular IL-2 expression (16,(29)(30)(31)(32), proliferation (11,33), and adenosine triphosphate (ATP) production (34)(35)(36).…”

mentioning

confidence: 99%

Whole blood flow cytometric measurement of NFATc1 and IL‐2 expression to analyze cyclosporine A‐mediated effects in T cells

et al. 2010

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The calcineurin inhibitor Cyclosporine A (CsA) is one of the crucial immunosuppressive drugs given after organ transplantation. The small therapeutic window of CsA generates the dilemma that efficient and toxic drug doses differ only slightly. Moreover, these threshold concentrations differ considerably between individuals; therefore, functional assays are urgently needed. We explored whether the transcription factor NFATc1, a direct as well as indirect target of CsA, can be used as a potential biomarker to determine the individual immunosuppressive activity of CsA. First, in isolated human T cells we showed that flow cytometry is practicable to measure NFATc1, the most abundant NFATc isoform in activated T cells. Second, for whole blood we developed a flow cytometric assay to determine in parallel the inducible transcription factor NFATc1 and the cytokine IL-2 in stimulated T cells. We found that added CsA inhibits both the expression of NFATc1 and IL-2 in T cells of stimulated whole blood samples with IC 50 values of 200 and 150 nM, respectively. The intra-and inter-assay variability was low, and clinical practicability was good. Further experiments have to demonstrate whether the parallel cytometric measurement of NFATc1 and IL-2 in whole blood is a good predictor of individual CsA efficacy and toxicity in CsA-treated patients. '

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Novel method for the measurement of cytokine production by a one-stage procedure

Cited by 57 publications

References 16 publications

Stimulation of whole blood cultures in patients with ankylosing spondylitis by a mitogen derived fromMycoplasma arthritidis (MAS) and other mitogens

Stimulation of whole blood cultures in patients with ankylosing spondylitis by a mitogen derived fromMycoplasma arthritidis (MAS) and other mitogens

Major Depression and Activation of The Inflammatory Response System

Whole blood flow cytometric measurement of NFATc1 and IL‐2 expression to analyze cyclosporine A‐mediated effects in T cells

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