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The ex vivo effects of macrophage colony-stimulating factor (M-CSF) on antifungal and antibacterial activities of human elutriated monocytes were studied. Cells were isolated prior to the initiation of therapy, on day 3 and at week 7, in six patients with an advanced malignancy receiving M-CSF in a phase I study. Superoxide anion production by monocytes in response to N-formyl methionyl leucyl phenylalanine...
We examined the mRNA levels for various cytokines, including IL-1α, IL-1β, TNFα, TGF-β 1 , GM-CSF, IL-6, IL-8, bFGF, PDGF-A, PDGF-B and IL-1ra, and IL-1β converting enzyme, and the protein levels of some of these cytokines in 19 SV40-transformed synovial cell clones. Among those tested, the mRNA levels for IL-6, bFGF and PDGF-A in rheumatoid arthritis (RA) cell clones were greater than those...
Interleukin(IL-)6 is overproduced in psoriatic lesions. We investigated the contribution of dermal fibroblasts to the local IL-6 production. Fibroblasts (passageno. 2 to 6) derived from lesional psoriatic (PP) and normal human (NN) skin were used to analyse the secretion of IL-6, and the related cytokines IL-1, IL-8 and TNF-α, and the expression of their corresponding mRNA by bioassay, ELISA and Northern...
The N-terminal amino acid sequence of the anti-neoplastic urinary protein (ANUP), a unique cytokine present in human granulocytes, was determined to be: Pyroglu-Leu-Lys-X-Tyr-Thr-X-Lys-Glu-Pro-Met-Thr-Ser(Thr)-Ala-Ala . . . This sequence showed no significant homology with any other protein when used in database searches. Furthermore, a synthetic nonapeptide corresponding to the first nine residues,...
We have developed a simple, rapid, sensitive bioassay for megakaryocyte growth and development factor (MGDF) or thrombopoietin (TPO) based on its ability to stimulate the proliferation of the human megakaryoblastic cell line MO7e. The bioassay takes only 24 h, is reproducible and sensitive to less than 10 pg/ml of MGDF. It can be made specific for MGDF by including neutralizing antibodies specific...
Recombinant chicken stem cell factor (SCF) was produced in bacteria as a histidine-tagged protein (HisΔSCF) and purified by affinity chromatography on an Ni 2+ -NTA agarose column. HisΔSCF is devoid of signal peptide and transmembrane domains and therefore mimics a soluble form of SCF. We demonstrate that the factor is biologically fully active in supporting sustained proliferation of...
Multiple cytokines and growth factors are present at sites of inflammation, and each of these can potentially influence the nature of the inflammatory response. Vascular endothelial cells (ECs) must integrate the signals generated by these multiple factors to effectively regulate the immune response and homeostasis. IL-6 and IL-8 are endothelial-derived products which play an important role as regulators...
Human ADF (adult T cell leukaemia-derived factor), an isoform of thioredoxin, promotes proliferation of certain human lymphoid cell lines and is involved in many thiol-dependent reducing reactions. To study functional aspects of the murine homologue, we established inducible overexpression of murine ADF inE. coliand a purification method which led to an apparently homogeneous 14 kDa protein. This...
The specific expression of pro-inflammatory cytokines may affect the functioning of organs in different ways. The results of specific cytokine bioassays used in this study show a distinct pattern of tissue expression of IL-1 IL-6 and CSF-1. Cytokine activity was assessed in conditioned media (CM) and lysates (LYS), obtained from different organs of control or lipopolysaccharide (LPS)-injected mice;...
Monocytes cultured in vitro differentiate to a macrophage-like phenotype and undergo functional changes, including reduced capacity for release of TNF-α and the soluble p55 receptor for TNF (sTNF-R55) but enhanced capacity for release of the soluble p75 receptor (sTNF-R75). The cytokines IL-4 and IL-10 act on monocytes to suppress the release of pro-inflammatory cytokines, including TNF-α, and to...
To investigate the spectrum of cytokines expressed by peripheral blood mononuclear cells (PBMC) from cynomolgus macaques (Macaca fascicularis), we used a semi-quantitative RT-PCR to determine levels of mRNA coding for IL-1β, IL-2, IL-4, IL-6, IL-10, IFN-γ, and TNF-α. The PCR products were labelled and quantified using a new fluorescent tag TOTO-1 (thiazole orange dimer) and an automated fluorescence-based...
Psoriatic fibroblasts produce enhanced amounts of IL-6 in vitro. This state of activation may reflect an altered expression of cytokine receptors, involved in auto/paracrine induction of IL-6. Cultures of dermal fibroblasts derived from lesional psoriatic (PP) and normal control (NN) skin were therefore analysed for their ability to bind biotinylated recombinant human cytokines using flow cytometry...
Monocyte cytotoxicity inducing factor (MCF) is a cytokine derived from CD4+ lymphocytes which was isolated using its ability to cause human blood monocytes to become cytotoxic for tumour targets. An N-terminal derived biologically active peptide (GAAVLEDSQ) of the intact molecule was previously used to demonstrate a single class of high affinity binding sites on human blood monocytes and U937 cells...
Pro-inflammatory cytokines are produced after systemic or local inflammation by a wide variety of cell types including monocytes, macrophages, Kupffer and endothelial cells. Previous studies have shown that IL-6 gene expression does not occur in liver from rats undergoing an acute phase response after turpentine injection or controls. These data do not rule out the possibility that delivery of a pathogen...
To investigate the physiological significance of hepatocyte growth factor (HGF) after trans-catheter arterial chemo-embolization (TACE), we measured serum HGF levels in 23 patients who underwent angiography (n= 3) or TACE (n= 20). HGF levels increased biphasically after TACE . It rapidly increased to 10-fold higher levels immediately after heparin-injection even before embolization. Serum HGF then...
A murine model of haemorrhagic shock was used to investigate bacterial translocation from the gut and subsequent systemic immunoreduction. Anaesthetized mice were bled from the femoral artery, and held at a mean arterial blood pressure of 35 mm Hg for one hour then resuscitated with shed blood and two-fold volume lactated Ringer's solution. Upon awakening, they were given cytokines or control media...
We have examined the lipopolysaccharide (LPS) induced release of interleukin 1β (IL-1β), IL-6, IL-8 and tumour necrosis factor alpha (TNF-α), in EDTA or heparin anticoagulated whole blood, from persons with high or low levels of high density lipoprotein (HDL), incubated at 37 o C for up to 20 h. In general, 100 ng LPS/ml gave two to twenty times higher responses than 1 ng LPS/ml, and heparin...
The anti-inflammatory properties of IL-4, IL-10, IL-13 and TGF-β are associated with their ability to repress the production of pro-inflammatory cytokines and to favour the release of interleukin-1 receptor antagonist (IL-1ra). Here, we investigate their actions on activated human polymorphonuclear cells (PMN). IL-4 and TGF-β were able to increase the production of IL-1ra, however only IL-4 were able...
Intraarticular injection of zymosan (1 mg) into the knee joints of male Wistar rats led to infiltration of neutrophils and monocytes, joint swelling and elevation of tumour necrosis factor (TNF-α) in joint lavage fluids. Neutrophil infiltration was maximal at 5 h after induction of arthritis but had declined by 24 h post injection. Monocyte infiltration was maximal around the same time as that of...
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