Determination of Chondroitin-6-Sulphate by a Competitive Enzyme Immunoassay Using a Biotinylated Antigen

Kähnert, Heike; Brinkmann, Thomas; Gässler, Norbert; Kleesiek, Knut

doi:10.1515/cclm.1994.32.4.293

Cited by 3 publications

(3 citation statements)

References 9 publications

(11 reference statements)

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“…Quantitation of chondroitin-6-sulfate Chondroitin-6-sulfate concentrations in cell lysates and cell culture medium were determined with a competitive enzyme immunoassay using biotinylated chondroitin-6-sulfate antigen as described before (Ka ¨hnert et al, 1994).…”

Section: Determination Of Xt Activitymentioning

confidence: 99%

Serum Xylosyltransferase: a New Biochemical Marker of the Sclerotic Process in Systemic Sclerosis

Götting

Sollberg

Kühn

et al. 1999

Journal of Investigative Dermatology

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UDP-D-xylose:proteoglycan core protein beta-D-xylosyltransferase (EC2.4.2.26) is the initial enzyme in the biosynthesis of chondroitin sulfate and dermatan sulfate proteoglycans in fibroblasts and chondrocytes. Secretion of xylosyltransferase into the extracellular space was determined in cultured human dermal fibroblasts. A more than 6-fold accumulation of xylosyltransferase activity in cell culture supernatant was observed (day 1, 0.6 microU per 106 cells; day 9, 4.1 microU per 106 cells); however, intracellular xylosyltransferase activity remained at a constant level (0.4 microU per 106 cells). Exposure of human chondrocytes to colchicine led to a 3-fold decreased level of xylosyltransferase and chondroitin-6-sulfate concentration in cell culture. Specific xylosyltransferase activity and chondroitin-6-sulfate concentration decreased in a concentration-dependent manner and in parallel in culture medium and accumulated 5-fold in cell lysates indicating that xylosyltransferase is secreted simultaneously into the extracellular space with chondroitin sulfate proteoglycans. Xylosyltransferase activities were determined in serum samples of 30 patients with systemic sclerosis. Xylosyltransferase activities in female (mean value 1.28 mU per liter, 90% range 1.10-1.55 mU per liter) and male patients (mean 1.39 mU per liter, 90% range 1.16-1. 57 mU per liter) with systemic sclerosis were significantly increased in comparison with blood donors of a corresponding age. Furthermore, xylosyltransferase activity was correlated with the clinical classification of systemic sclerosis. Female patients with diffuse cutaneous systemic sclerosis showed higher serum xylosyltransferase activities than patients with limited systemic sclerosis. These results confirm that the increase of proteoglycan biosynthesis in sclerotic processes of scleroderma is closely related to an elevated xylosyltransferase activity in blood and demonstrate the validity of xylosyltransferase as an additional diagnostic marker for determination of sclerotic activity in systemic sclerosis.

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Section: Determination Of Xt Activitymentioning

confidence: 99%

Serum Xylosyltransferase: a New Biochemical Marker of the Sclerotic Process in Systemic Sclerosis

Götting

Sollberg

Kühn

et al. 1999

Journal of Investigative Dermatology

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“…In solid tumors such as colonic tumors, the stromata are rich in decorin (Adany et al, 1990) or CS (Wegrowski and Maquart, 2004). The overall CS‐concentration in human blood serum ranges between 5 and 30 µg/ml (Baici et al, 1992), and the concentration of CS‐6 sulfate (CS‐6S) was found to range between 55 and 169 ng/L in sera of healthy blood donors (Kähnert et al, 1994). The CS chains interact with a large number of ligands and receptors, which activate signaling pathways (Taylor and Gallo, 2006) and stimulate tumor growth.…”

mentioning

confidence: 99%

Decorin and chondroitin‐6 sulfate inhibit B16V melanoma cell migration and invasion by cellular acidification

Stock¹,

Jungmann

Seidler

2011

Journal Cellular Physiology

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In vivo, cells are embedded in an environment generated and maintained by multiple cell-cell and cell-matrix interactions. While transiting the dermis metastasizing melanoma cells interact with the extracellular matrix (ECM) and fibroblasts. To study the roles of ECM components and fibroblasts in melanoma (B16V) cell migration and invasion, we established a co-culture system consisting of fibroblasts, their collagen-rich matrix and B16V cells. The crosstalk between B16V cells and fibroblasts was indicated by a clear increase in release and activity of matrix-metallo-protease-2. Time-lapse microscopy revealed that in B16V cells exposed to either decorin or chondroitin sulfates migration and invasion decreased by more than 50%. Decorin led to a reversible, chondroitin-6-sulfate to an irreversible, cytosolic acidification of B16V cells. Interestingly, decorin lowered NHE1 activity whereas chondroitin-6-sulfate did not. Furthermore, decorin and chondroitin-6-sulfate also acidified the pH at the cell surface which might prevent migration due to strong adhesion. In conclusion, the present co-culture system is an appropriate tool to analyze migration, invasion, and MMP release depending on cell-matrix interactions and the crosstalk between the invasive cells and those surrounded by their self-made matrix. We show a so far unknown function of decorin and chondroitin-6-sulfate: their ability to inhibit B16V cell migration by intracellular acidification.

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“…All chemicals used were of analytical grade or the highest grade commercially available. phate (9), hyaluronan) and markers specific for macrophages (neopterin, sCD14) (data not shown).…”

Section: Methodsmentioning

confidence: 89%

Synthesis of Tissue Factor Pathway Inhibitor in Human Synovial Cells and Chondrocytes Makes Joints the Predilected Site of Bleeding in Haemophiliacs

Brinkmann

Kähnert²,

Prohaska³

et al. 1994

CCLM

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Summary:The synthesis of tissue factor pathway inhibitor (TFPI) was investigated in cloned human synovial cells and human chondrocytes. TFPI-specific DNA transcription products of these cells were isolated, and a full-length cDNA of about 1000 base pairs was amplified by reverse transcription and polymerase chain reaction. The amplified DNA was cloned into the vector pUC 18. The TFPI coding sequence was confirmed by double stranded sequencing and was identical with that previously published for human TFPI coding nucleotide sequence frorn human placental cDNA (1).The inhibitory activity of TFPI in the cell medium of cultivated human chondrocytes and cloned human synovial cells was determined by a specific chromogenic Substrate assay of factor Xa activity. The inhibitory activity of TFPI in the medium of human chondrocytes and cloned human synovial cells was 630-720 mU/10 8 cells and 1080-1665 mU/10 8 cells, respectively.In addition, TFPI activity in cell culture media of human chondrocytes and cloned human synovial cells was suppressed by a polyclonal goat anti-TFPI antibody directed against the inhibitory domain I and domain II. In the chromogenic Substrate assay, the anti-TFPl antibody completely suppressed the inhibitory activity of TFPI in the samples.

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Determination of Chondroitin-6-Sulphate by a Competitive Enzyme Immunoassay Using a Biotinylated Antigen

Cited by 3 publications

References 9 publications

Serum Xylosyltransferase: a New Biochemical Marker of the Sclerotic Process in Systemic Sclerosis

Serum Xylosyltransferase: a New Biochemical Marker of the Sclerotic Process in Systemic Sclerosis

Decorin and chondroitin‐6 sulfate inhibit B16V melanoma cell migration and invasion by cellular acidification

Synthesis of Tissue Factor Pathway Inhibitor in Human Synovial Cells and Chondrocytes Makes Joints the Predilected Site of Bleeding in Haemophiliacs

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