Production of tumour necrosis factor-α by cultured human peripheral blood leucocytes in response to the anti-tumour agent 5,6-dimethylxanthenone-4-acetic acid (NSC 640488)

Background:In our previous study we found that rhsIL-6R, along with recombinant human interleukin-6, plays a regulatory role in the immune response by modulating the tumour necrosis factor-α (TNF-α) expression and its production by peripheral blood mononuclearcells (PBMC). We also suggested that sIL-6R with IL-6 secreted by human PMN (neutrophils) influenced the TNF-α expression and its production by autologous PBMC.Aims:Since soluble gp130 (sgp130) is a natural inhibitor for sIL-6R/interleukin-6 responses, in the present study we estimated an effect of exogenous recombinant human sgp130 and sgp130 secreted by PMN on the TNF-α expression and its production by PBMC.Methods:Cells were isolated from whole blood of healthy persons. The PMN were cultured in 96-well plates for 1 h at 37°C in a humidified incubator with 5% CO2. After the incubation, the culture supernatant of PMN was removed and added to the PBMC. PBMC were incubated for 1 h at 37°C in the same conditions. Cytoplasmic protein fractions of PMN and, for comparative purpose of PBMC, were analysed for presence of sgp130 by western blotting with the use of monoclonal antibody capable of detecting this protein. In the culture supernatants of PMN we examined the concentrations of sgp130 by human enzyme-linked immunosorbent assay. TNF-α was measured at the protein levels as well as the mRNA levels.Results and conclusions:The present results revealed that exogenous recombinant human sgp130 modulates the TNF-α expression and production by PBMC. In contrast, we did not find any effect of sgp130 secreted by PMN on the TNF-α expression and its production by autologous PBMC.

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Immunomodulatory and Anti-Inflammatory Activity of Selected Osthole Derivatives

Zeitschrift für Naturforschung C ◽

10.1515/znc-2009-5-610 ◽

2009 ◽

Vol 64 (5-6) ◽

pp. 361-368 ◽

Cited By ~ 32

Author(s):

Michał Zimecki ◽

Jolanta Artym ◽

Wojciech Cisowski ◽

Irena Mażol ◽

Maciej Włodarczyk ◽

...

Keyword(s):

Tumour Necrosis Factor ◽

Interleukin 6 ◽

Tumour Necrosis ◽

Mononuclear Cells ◽

Human Peripheral Blood ◽

Pokeweed Mitogen ◽

Tumour Necrosis Factor Α ◽

Anti Inflammatory ◽

Factor Α ◽

Necrosis Factor

IFrom osthole [7-methoxy-8-(3-methyl-but-2-enyl)-chromen-2-one] (), obtained by selective extraction of Peucedanum ostruthium (L.) W. Koch roots, ostholic acid (II) was synthetized as a result of its oxidation with chromium trioxide. From ostholic acid, through its chloride, four amides were obtained: the morpholide 1, the p-chloro-benzylamide 2, the piperidine 3 and the N-methyl-piperazide 4. Except for 1, other compounds have not been described before. The amides 1 - 4 and their precursor osthole (I) were tested for their potential activities in selected immunological assays. The compounds showed moderate inhibitory activity in the humoral immune response to sheep erythrocytes in mice in vitro, and 4 was the most suppressive. The effects of 1 and 3 on concanavalin A- and pokeweed mitogen-induced mouse splenocyte proliferation were inhibitory and those of 4 stimulatory. The compounds were also tested for their activity on tumour necrosis factor α and interleukin 6 production, induced by lipopolysaccharide, in cultures of rat peritoneal cells and human peripheral blood mononuclear cells. Compounds 1, 3 and 4 inhibited tumour necrosis factor α (rat cells), whereas compound 2 stimulated the production of both cytokines. Compounds 1, 2 and 3 were also strongly inhibitory on tumour necrosis factor α production in human blood cells (73, 78 and 80% inhibition at 10 μg/ml, respectively). On the other hand, 2 and 4 stimulated the interleukin 6 production (2- to 3-fold stimulation). In addition, 2 and 4 suppressed the carrageenan-induced inflammation in mice (56.5% and 68.3% inhibition, respectively). In summary, the compounds predominantly displayed suppressive and anti inflammatory activities in the investigated models

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Activated tumour necrosis factor-α shedding process is associated with in-hospital complication in patients with acute myocardial infarction

Clinical Science ◽

10.1042/cs20040229 ◽

2005 ◽

Vol 108 (4) ◽

pp. 339-347 ◽

Cited By ~ 20

Author(s):

Yudai SHIMODA ◽

Mamoru SATOH ◽

Motoyuki NAKAMURA ◽

Tomonari AKATSU ◽

Katsuhiko HIRAMORI

Keyword(s):

Myocardial Infarction ◽

Acute Myocardial Infarction ◽

Tumour Necrosis Factor ◽

Peripheral Blood ◽

Tumour Necrosis ◽

Mrna Levels ◽

Tumour Necrosis Factor Α ◽

Tnf Α ◽

Factor Α ◽

Necrosis Factor

TACE [TNF-α (tumour necrosis factor-α)-converting enzyme] plays an essential role in the shedding of TNF-α, which could affect the outcome of AMI (acute myocardial infarction). To investigate the clinical significance of the TACE–TNF-α system in AMI, we examined TACE-mediated TNF-α synthesis in PBMCs (peripheral blood mononuclear cells), which are a possible source of TNF-α in AMI. Forty-one patients with AMI and 15 healthy subjects (HS) were enrolled in the present study. PBMCs were isolated from peripheral blood on day 1 and 14 after the onset of AMI. TACE and TNF-α mRNA levels and intracellular median fluorescence intensity were measured by real-time RT (reverse transcriptase)–PCR and flow cytometry respectively. TACE-mediated TNF-α production was evaluated in cultured PBMCs with PMA, which is known to activate TACE. Spontaneous TACE and TNF-α levels were higher in AMI patients than in HS (P<0.001). TACE and TNF-α levels in PMA-stimulated PMBCs were markedly increased in AMI patients compared with HS (P<0.001). There was a positive correlation between TACE and TNF-α levels in AMI. Although spontaneous and stimulated levels of TACE and TNF-α decreased 14 days after the onset of AMI, levels in AMI patients were higher than in HS. In AMI patients with in-hospital complications (n=15; pump failure in ten, recurrent myocardial infarction in one, malignant ventricular arrhythmia in three and cardiac death in one), spontaneous and stimulated levels of TACE and TNF-α were higher than in patients without complications (P<0.01). These levels were higher in AMI patients with in-hospital complications 14 days after onset. These results demonstrate that TACE-mediated TNF-α maturation in PBMCs may play an important role in poor outcomes from AMI, suggesting that TACE may be a potential target for the inhibition of cellular TNF-α production in AMI.

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