Sensitive ELISA for interleukin-6

Helle, M.; Boeije, L. C. M.; Groot, Els de; Vos, Anneke S. de; Aarden, Lucien A.

doi:10.1016/0022-1759(91)90063-l

Cited by 213 publications

(19 citation statements)

References 23 publications

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“…TNF and IL-6 levels were determined using an immunoradiometric assay kit (IRE Medgenix, Bruxelles, Belgium) and an ELISA (14), respectively. Routine hematologic and coagulation tests were performed using standard laboratory methods.…”

Section: Methodsmentioning

confidence: 99%

Inhibition of endotoxin-induced activation of coagulation and fibrinolysis by pentoxifylline or by a monoclonal anti-tissue factor antibody in chimpanzees.

Levi¹,

Cate²,

Bauer³

et al. 1994

J. Clin. Invest.

412

243

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Introduction Knowledge of the pathogenetic mechanisms responsible for the activation of the coagulation system associated with endotoxemia is important for the development of improved modalities for prevention and treatment. We analyzed the appearance in plasma of TNF, IL-6, and indices of coagulation and fibrinolytic system activation in normal chimpanzees after intravenous infusion of endotoxin.Endotoxin infusion elicited reproducible and dose-dependent elevations in serum TNF and IL-6, as well as marked increases in thrombin generation in vivo as measured by immunoassays for prothrombin activation fragment F1+2, thrombinantithrombin III complexes, and fibrinopeptide A. Activation of the fibrinolytic mechanism was monitored with assays for plasminogen activator activity and plasmin-a2-antiplasmin complexes. To potentially intervene in the molecular pathways elicited by endotoxin, pentoxifylline, an agent that interrupts "immediate early" gene activation by monocytes, or a potent monoclonal antibody that neutralizes tissue factor-mediated initiation of coagulation, were infused shortly before endotoxin. Pentoxifylline markedly inhibited increases in the levels of TNF and IL-6, as well as the effects on coagulation and fibrinolysis. In contrast, the monoclonal antibody to tissue factor completely abrogated the augmentation in thrombin generation, but had no effect on cytokine levels or fibrinolysis.We conclude that the endotoxin-induced activation ofcoagulation appears to be mediated by the tissue factor-dependent pathway, the fibrinolytic response triggered by endotoxin is not dependent on the generation of thrombin, and that the release of cytokines may be important in mediating the activation of both the coagulation and the fibrinolytic mechanisms in vivo. (J. Clin. Invest. 1994.93:114-120.)

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Section: Methodsmentioning

confidence: 99%

Inhibition of endotoxin-induced activation of coagulation and fibrinolysis by pentoxifylline or by a monoclonal anti-tissue factor antibody in chimpanzees.

Levi¹,

Cate²,

Bauer³

et al. 1994

J. Clin. Invest.

412

243

View full text Add to dashboard Cite

show abstract

“…ShIL6R expressed in baculovirus-infected insect cells [23] was used as a standard. IL-6 was quantified by a sandwich ELISA as described [25].…”

Section: Methodsmentioning

confidence: 99%

Purification and Characterization of the Soluble Interleukin‐6 Receptor from Human Plasma and Identification of An Isoform Generated through Alternative Splicing

Müller‐Newen

Köhne

Keul

et al. 1996

European Journal of Biochemistry

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The soluble human interleukin-6 receptor (shIL6R) was purified from human plasma. In a single immunoaffinity purification step a 140000-fold enrichment with a yield of 95% was achieved. A subsequent IL-6 affinity chromatography resulted in a homogeneous receptor preparation but only in a yield of less than 5%. The biological activity of the soluble receptor was clearly demonstrated by its ability to induce the synthesis of the acute-phase protein a, -antichymotrypsin in HepG2 cells stably transfected with IL-6. Upon gel filtration, the native shIL6R showed an apparent molecular mass of 93 kDa. Analysis by SDSPAGE revealed an apparent molecular mass of 65 kDa for the soluble receptor. Deglycosylation with peptide N-glycosidase F led to a shift in molecular mass from 65 kDa to 45 kDa. It has previously been shown that the shIL6R can be generated by shedding the membrane-bound form or by expression of an alternatively spliced mRNA. Here we show that the shIL6R isolated from human plasma is recognized by an affinity-purified peptide antibody raised against an amino acid sequence unique for the alternatively spliced isoform. Thus, the shIL6R isoform generated through alternative splicing which has been previously detected in supernatants of cultured cell lines is also an in vivo product circulating in human plasma.

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“…Procedures were identical to the IL-12 ELISA, except that the blocking step was eliminated and for coating anti-IL-6, mAb CLB.IL6/16 was used at 1 g/ml in PBS. Affinity-purified biotinylated polyclonal sheep anti-IL-6 was used for detection at 0.25 g/ ml (33). rhIL-6 (34) was used as a standard.…”

Section: Assays For Cytokinesmentioning

confidence: 99%

Histamine inhibits the production of interleukin-12 through interaction with H2 receptors.

Kraan¹,

Snijders²,

Boeije³

et al. 1998

J. Clin. Invest.

190

113

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Sensitive ELISA for interleukin-6

Cited by 213 publications

References 23 publications

Inhibition of endotoxin-induced activation of coagulation and fibrinolysis by pentoxifylline or by a monoclonal anti-tissue factor antibody in chimpanzees.

Inhibition of endotoxin-induced activation of coagulation and fibrinolysis by pentoxifylline or by a monoclonal anti-tissue factor antibody in chimpanzees.

Purification and Characterization of the Soluble Interleukin‐6 Receptor from Human Plasma and Identification of An Isoform Generated through Alternative Splicing

Histamine inhibits the production of interleukin-12 through interaction with H2 receptors.

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