St. Georg, Germany
St. Georg, Germany

Time filter

Source Type

Raspe C.,Martin Luther University of Halle Wittenberg | Czeslick E.,Critical Care and Pain Therapy | Leimert A.,Martin Luther University of Halle Wittenberg | Rossberg T.,St Georg Medical Center | And 4 more authors.
International Journal of Interferon, Cytokine and Mediator Research | Year: 2013

Objective: The study reported here aimed to analyze the anti-inflammatory and immunmodulatory impact of arginine, glycine, glutamine, and the combination of these amino acids on the intracellular expression of proinflammatory cytokines during sepsis. These amino acids were tested on lipopolysaccharide (LPS)-stimulated human monocytes in a whole-blood system and examined using flow cytometry. Materials and methods: The whole blood of twelve healthy volunteers processed immediately after withdrawal was incubated with arginine (2 and 5 mM), glycine (2 and 5 mM), glycyl-glutamine (2 and 5 mM), and the amino acid and dipeptide solution Glamin® (Fresenius Kabi, Bad Homburg, Germany) at three concentrations (5%, 10%, 20%), with or without LPS (0.2 ng/mL) stimulation. Cytokine-producing monocytes were phenotyped in whole blood and the intracellular expression of interleukin (IL)-6, IL-8, and tumor necrosis factor (TNF)-α was assessed by flow cytometry. Results: In whole-blood samples from volunteers, used to best imitate physiological cellular interactions, the amino acid and dipeptide solution Glamin, containing arginine, glycine, glycyl-glutamine, was able to significantly and dose-dependently diminish LPS-induced production of proinflammatory cytokines such as IL-6, IL-8, and TNF-α in human monocytes. However, single incubation with the amino acids arginine, glycine, and glycyl-glutamine individually did not affect alterations in the expression of IL-6, IL-8, or TNF-α. Conclusion: The amino acid and dipeptide solution Glamin, composed of glycine, arginine, and glycyl-glutamine, had strong anti-inflammatory and immunomodulatory effects during the induced experimental sepsis, as it significantly downregulated intracellular expression of proinflammatory cytokines in human whole-blood monocytes. However, only incubation with a combination of amino acids (ie, Glamin), rather than individual amino acids, demonstrated an inhibitory impact on cytokine production in LPS-stimulated monocytes. The results indicate that a combination of amino acids may potentiate the anti-inflammatory response, leading to a marked suppression of TNF-α, IL-6, and IL-8 during sepsis. © 2013 Raspé et al, publisher and licensee Dove Medical Press Ltd.


Raspe C.,Martin Luther University of Halle Wittenberg | Czeslick E.,St Georg Medical Center | Weimann A.,St Georg Medical Center | Schinke C.,Jacobi Medical Center | And 5 more authors.
Cytokine | Year: 2013

To investigate the effects of the commonly-used immunomodulators l-glutamine, l-alanine, and the combination of both l-alanyl-l-glutamine (Dipeptamin®) on intracellular expression of IL-6, IL-8, and TNF-α during endotoxemia, lipopolysaccharide (LPS)-stimulated human monocytes in a whole blood system were investigated by flow cytometry. Whole blood of twenty-seven healthy volunteers was stimulated with LPS and incubated with three different amino acid solutions (1. l-glutamine, 2. l-alanine, 3. l-alanyl-l-glutamine, each concentration 2mM, 5mM, incubation time 3h). CD14+ monocytes were phenotyped in whole-blood and intracellular expression of cytokines was assessed by flow cytometry. Our investigations showed for the first time in whole blood probes, imitating best physiologically present cellular interactions, that l-glutamine caused a dose-independent inhibitory effect on IL-6 and TNF-α production in human monocytes stimulated with LPS. However, l-alanine had contrary effects on IL-6 expression, significantly upregulating expression of IL-6 in LPS-treated monocytes. The impact of l-alanine on the expression of TNF-α was comparable with glutamine. Neither amino acid was able to affect IL-8 production in LPS-stimulated monocytes. The combination of both did not influence significantly IL-6 and IL-8 expression in monocytes during endotoxemia, however strongly reduced TNF-α production. For the regulation of TNF-α, l-glutamine, l-alanine and the combination of both show a congruent and exponentiated downregulating effect during endotoxemia, for the modulation of IL-6, l-glutamine and l-alanine featured opposite regulation leading to a canceling impact of each other when recombining both amino acids. © 2013 Elsevier Ltd.


Kellner P.,Martin Luther University of Halle Wittenberg | Nestler F.,Martin Luther University of Halle Wittenberg | Leimert A.,Martin Luther University of Halle Wittenberg | Bucher M.,Martin Luther University of Halle Wittenberg | And 4 more authors.
Cytokine | Year: 2014

In order to examine the immunomodulatory effects of antithrombin III (AT-III) and C1 esterase inhibitor (C1-INH) in human monocytes, we investigated the intracellular expression of interleukin (IL)-6, IL-8, and tumor necrosis factor (TNF)-α in an ex-vivo laboratory study in a whole blood setting. Heparinized whole blood samples from 23 healthy male and female volunteers (mean age: 27±7years) were pre-incubated with clinically relevant concentrations of AT-III (n=11) and C1-INH (n=12), then stimulated with 0.2ng/mL lipopolysaccharide (LPS) for 3h. After phenotyping CD14+ monocytes, intracellular expression of IL-6, IL-8, and TNF-α was assessed using flow cytometry. In addition, 12 whole blood samples (AT-III and C1-INH, n=6 each) were examined using hirudin for anticoagulation; all samples were processed in the same way. To exclude cytotoxicity effects, 7-amino-actinomycin D and Nonidet P40 staining were used to investigate probes. This study is the first to demonstrate the influence of C1-INH and AT-III on the monocytic inflammatory response in a whole blood setting, which mimics the optimal physiological setting. Cells treated with AT-III exhibited significant downregulation of the proportion of gated CD14+ monocytes for IL-6 and IL-8, in a dose-dependent manner; downregulation for TNF-α did not reach statistical significance. There were no significant effects on mean fluorescence intensity (MFI). In contrast, C1-INH did not significantly reduce the proportion of gated CD14+ monocytes or the MFI regarding IL-6, TNF-α, and IL-8. When using hirudin for anticoagulation, no difference in the anti-inflammatory properties of AT-III and C1-INH in monocytes occurs. Taken together, in contrast to TNF-α, IL-6 and IL-8 were significantly downregulated in monocytes in an ex-vivo setting of human whole blood when treated with AT-III. This finding implicates monocytes as an important point of action regarding the anti-inflammatory properties of AT-III in sepsis. C1-INH was unable to attenuate the monocytic response, which supports the hypothesis that other cellular components in whole blood (e.g., neutrophils) might be responsible for the known effects of C1-INH in inflammation. © 2014 Elsevier Ltd.


PubMed | Martin Luther University of Halle Wittenberg and St Georg Medical Center
Type: Journal Article | Journal: Cytokine | Year: 2014

In order to examine the immunomodulatory effects of antithrombin III (AT-III) and C1 esterase inhibitor (C1-INH) in human monocytes, we investigated the intracellular expression of interleukin (IL)-6, IL-8, and tumor necrosis factor (TNF)- in an ex-vivo laboratory study in a whole blood setting. Heparinized whole blood samples from 23 healthy male and female volunteers (mean age: 277years) were pre-incubated with clinically relevant concentrations of AT-III (n=11) and C1-INH (n=12), then stimulated with 0.2 ng/mL lipopolysaccharide (LPS) for 3h. After phenotyping CD14 monocytes, intracellular expression of IL-6, IL-8, and TNF- was assessed using flow cytometry. In addition, 12 whole blood samples (AT-III and C1-INH, n=6 each) were examined using hirudin for anticoagulation; all samples were processed in the same way. To exclude cytotoxicity effects, 7-amino-actinomycin D and Nonidet P40 staining were used to investigate probes. This study is the first to demonstrate the influence of C1-INH and AT-III on the monocytic inflammatory response in a whole blood setting, which mimics the optimal physiological setting. Cells treated with AT-III exhibited significant downregulation of the proportion of gated CD14 monocytes for IL-6 and IL-8, in a dose-dependent manner; downregulation for TNF- did not reach statistical significance. There were no significant effects on mean fluorescence intensity (MFI). In contrast, C1-INH did not significantly reduce the proportion of gated CD14 monocytes or the MFI regarding IL-6, TNF-, and IL-8. When using hirudin for anticoagulation, no difference in the anti-inflammatory properties of AT-III and C1-INH in monocytes occurs. Taken together, in contrast to TNF-, IL-6 and IL-8 were significantly downregulated in monocytes in an ex-vivo setting of human whole blood when treated with AT-III. This finding implicates monocytes as an important point of action regarding the anti-inflammatory properties of AT-III in sepsis. C1-INH was unable to attenuate the monocytic response, which supports the hypothesis that other cellular components in whole blood (e.g., neutrophils) might be responsible for the known effects of C1-INH in inflammation.

Loading St Georg Medical Center collaborators
Loading St Georg Medical Center collaborators