Enzyme-Linked Immunoassay for Human Granulocyte Elastase in Complex with α1 -Proteinase Inhibitor

Neumann, Siegfried; Hennrich, Norbert; Gunzer, Gerhard; Lang, Hermann

doi:10.1007/978-1-4615-9355-3_33

Cited by 69 publications

(32 citation statements)

References 10 publications

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“…In our hands, the HLE levels in control plasma were compatible with the previously published values of 84 _+ 25 ng/ml (20) and 60 ng/ml (median value) (24) using the same assay system. The majority of the plasma samples from RA patients had clearly elevated HLE-a PI levels (Fig.…”

Section: Resultssupporting

confidence: 91%

“…The design of the enzyme-linked immunoassay (20) ensures that only HLE bound to a -proteinase inhibitor was measured (Method A). In our hands, the HLE levels in control plasma were compatible with the previously published values of 84 _+ 25 ng/ml (20) and 60 ng/ml (median value) (24) using the same assay system.…”

Section: Resultsmentioning

confidence: 99%

“…Levels of immunreactive elastase in plasma were determined with a newly developed enzyme-linked immunoassay (ELISA) (12,20). This method (Method A) measures exclusively the elastase complexed to a^PI.…”

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

Plasma Levels of Inhibitor-Bound Leukocytic Elastase in Rheumatoid Arthritis Patients

Schnebli

Christen

Jochum

et al. 1984

Advances in Experimental Medicine and Biology

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Section: Resultssupporting

confidence: 91%

Section: Resultsmentioning

confidence: 99%

See 1 more Smart Citation

Plasma Levels of Inhibitor-Bound Leukocytic Elastase in Rheumatoid Arthritis Patients

Schnebli

Christen

Jochum

et al. 1984

Advances in Experimental Medicine and Biology

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“…The justification for the assay method using this kit has been reported by one of the co-authors (Ikei et al, 1992). This is a sensitive assay that enables rapid measurement of both PMN-E-complexed al-proteinase inhibitor and the free form of PMN-E, which may be present in the enzyme samples, in contrast to the conventional Merck EIA kit (E. Merck, Darmstadt, Germany), which detects only PMN-E complexed with xl-proteinase inhibitor (Neumann et al, 1984). When 0.1 ml of tissue extract was used, the detection limit of ir-PMN-E was 0.063 1ig per 100 mg of protein.…”

Section: Cell Culturementioning

confidence: 99%

Production of immunoreactive polymorphonuclear leucocyte elastase in human breast cancer cells: possible role of polymorphonuclear leucocyte elastase in the progression of human breast cancer

Yamashita

Ogawa²,

Ikei³

et al. 1994

Br J Cancer

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SummaryBreast cancer cells are known to express various proteolytic enzymes, which make them invasive and favour their dissemination to distant sites. However, it is unclear whether breast cancer cells have the ability to produce polymorphonuclear leucocyte elastase (PMN-E). We measured immunoreactive (ir) PMN-E content in the conditioned medium of two breast cancer cell lines, MCF-7 and ZR-75-1, and two normal breast epithelial cell lines, HBL-100 and Hs 578Bst, using a highly specific and sensitive enzyme immunoassay. (4.23 ± 3.74 gg 100 mg-' protein) patients. Breast cancer patients with high levels of ir-PMN-E showed significantly shorter disease-free survival and overall survival than those with low levels of ir-PMN-E at the cut-off point of 8.99 lg 100 mg-' protein. In the multivariate analysis, ir-PMN-E content was found to be a significant prognostic factor for disease recurrence and death in human breast cancer.Considerable evidence suggests that proteolytic enzymes are involved in cancer invasion and metastasis (Mignatti et al., 1986;Persky et al., 1986). Production of tumour cell proteinases, including plasminogen activator (Duffy et al., 1987;Reich et al., 1988;Yamashita et al., 1993a), collagenase (Turpeenniemi-Hujanen et al., 1985), and cathepsin B (Recklies et al., 1980) has been implicated in tumour cell invasion into adjacent tissues and metastasis. Another proteinase that has attracted attention as a mediator of these processes is elastase. Elastase is the only proteinase that is able to degrade insoluble elastin (Janoff & Schere, 1968), a structural component of elastic tissues such as blood vessels, skin, lung and breast tissue.An elastinolytic activity has been demonstrated previously by Hornebeck et al. (1977) in tissue extracts from human breast cancer, but in their study it was not determined whether the activity could be attributed to tumour cells. Thereafter, several investigators have reported that elastinolytic activity is found in the culture medium of mouse mammary tumour cells (Grant et al., 1990;Zeydel et al., 1986) and human breast cancer cells (Kao et al., 1982), suggesting that elastinolytic enzymes are produced by the tumour cells themselves.Very recently, a highly specific and sensitive enzyme immunoassay (EIA) was established for the measurement of human polymorphonuclear leucocyte elastase (PMN-E) (Ikei et al., 1992). Labelled antibody in this assay recognises not only PMN-E complexed a,-proteinase inhibitor but also the free form of PMN-E, which may present in the enzyme samples. In the present study, we examined whether breast cancer cells have the ability to produce immunoreactive (ir)-PMN-E. Furthermore, we have determined the concentration of ir-PMN-E in tissue extracts from human breast cancer, and elucidated the relationship between the tissue content of ir-PMN-E and clinicopathological status in human breast cancer. Materials and methods Cell cultureThe human breast cancer cell line MCF-7 was kindly provided by Y. Nomura (National Kyushu Cancer Center Hospital, Fu...

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“…Plasma concentrations of alpha2-macroglobulin (o^M) and alphas-protease inhibitor ('oqPI) were evaluated by a radial immunodiffusion technique with standardized immunodiffusion plates (Behringwerke, Marburg, FRG). Quantitative estimation of the plasma levels of the elastase-alphaiprotease inhibitor (E-^PI) complex was carried out with a highly sensitive enzyme-linked immunoassay 9 . We report here the results obtained in: 12 healthy subjects aged from 22 -39 years (seven males, five females), 12 chronically uremic patients aged from 22 -66 years undergoing regular dialysis therapy (RDT) for 41.3 + 11.6 month (nine males, three females) and 10 patients with posttraumatic ARF aged from 19 -45 years.…”

Section: Methodsmentioning

confidence: 99%

Proteolytic Activity in Patients with Hypercatabolic Renal Failure

Hörl

Schäfer

Scheidhauer

et al. 1984

Advances in Experimental Medicine and Biology

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Enzyme-Linked Immunoassay for Human Granulocyte Elastase in Complex with α1 -Proteinase Inhibitor

Cited by 69 publications

References 10 publications

Plasma Levels of Inhibitor-Bound Leukocytic Elastase in Rheumatoid Arthritis Patients

Plasma Levels of Inhibitor-Bound Leukocytic Elastase in Rheumatoid Arthritis Patients

Production of immunoreactive polymorphonuclear leucocyte elastase in human breast cancer cells: possible role of polymorphonuclear leucocyte elastase in the progression of human breast cancer

Proteolytic Activity in Patients with Hypercatabolic Renal Failure

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