Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or...
Patent
1995-12-14
1998-06-16
Leary, Louise
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
435326, 435 721, 435 724, 435 72, 435 4052, 435 405, 435 71, 436 63, 436803, 128668, 128670, 128672, 128749, C12Q 100, G01N 3353
Patent
active
057668388
DESCRIPTION:
BRIEF SUMMARY
The present invention relates to a leukocyte-adhesion assay, more particularly a method of assaying leukocyte binding to vascular tissue.
Atherosclerosis is a disease of the arteries which results in approximately half of the total deaths in Western countries. The disease involves thickening of the inner part (intima) of the artery wall by infiltration of monocytes (a type of leukocyte normally present in the blood). Following infiltration, the monocytes mature into large cells called macrophages, which is followed by the accumulation of cholesterol, first within the macrophages but later becoming more widespread. Smooth muscle cells also appear and proliferate, probably under the influence of growth factors released by the macrophages. All of these components contribute to the thickening of the artery wall reducing the passage of blood through the artery which may finally become blocked by a blood clot forming at the site.
In recent years much has been learned about the mechanisms by which monocytes and other types of leukocytes migrate from the blood into tissues suffering from inflammation. One such mechanism involves adhesion molecules present on the inner surface of the vessels at sites of inflammation. The inner surface of all blood vessels consists of thin delicate cells referred to as endothelial cells and these form protein adhesion molecules which are able to bind specifically to receptors on leukocytes. These receptors are themselves adhesion molecules and consist of either carbohydrates, proteins or sulphur compounds on the surface of the leukocyte. Leukocytes can also be bound by other cells and structures within tissues by a number of similar adhesion interactions.
Recent work has shown that certain adhesion molecules are present on the endothelial cells of the diseased parts of atherosclerotic arteries but not in the normal parts of these arteries (see Poston et al, Am. J. Pathol., 140(3), 665-673 (1992)). One adhesion molecule (ICAM-1) also appears on smooth muscle cells within the thickened area but is not seen on these cells elsewhere. These observations suggest that atherosclerotic lesions may resemble foci of inflammation and that monocytes leave the blood under the influence of adhesion molecules entering and adding to the disease process.
Whatever the precise mechanism, it is clear that the adhesion of leukocytes, and particularly monocytes, to the arterial wall plays an important role in the development of atherosclerotic lesions. For this reason, there is a need for a method of assaying leukocyte binding to vascular tissue.
One method which has previously been used for studying the binding of leukocytes to tissue is the so-called Stamper-Woodruff assay. This assay was developed originally to study the mechanism of lymphocytes binding to lymph nodes (Stamper & Woodruff, J. Exp. Med., 144, 828-833 (1976)). A suspension of rat thoracic duct lymphocytes in RPMI medium was contacted at about 7.degree. C. with thin sections of rat or mouse lymph node tissue, bound cells were fixed and visualised and then identified under a light microscope.
The assay has subsequently been used in other applications and, for example, a modified version was used to investigate experimental autoimmune encephalomyelitis which is an inflammatory condition of the nervous system with similarities to multiple sclerosis (Yednock et al, Nature, 356, 63-66 (1992)). A suspension of a human monocytic cell line (U937) in RCMI medium was contacted with thin sections of brain tissue from normal rats and rats suffering from experimental autoimmune encephalomyelitis, bound cells were fixed and visualised and then identified under a light microscope. The brain tissue was generally contacted with U937 cells on ice although there is reference to some experiments having been carried out at 25.degree. C.
It has now been found that a version of the Stamper-Woodruff assay can be applied to provide a functional assay for leukocyte adhesion to vascular tissue, for example tissue derived from atherosclerotic lesions.
The present inv
REFERENCES:
patent: 4925678 (1990-05-01), Ranney
patent: 4935234 (1990-06-01), Todd, III et al.
patent: 5108759 (1992-04-01), Ranney
patent: 5166133 (1992-11-01), Houston et al.
patent: 5217870 (1993-06-01), Hession et al.
patent: 5503982 (1996-04-01), Hendricks et al.
patent: 5514555 (1996-05-01), Springer et al.
DiCoreleto et al., J. Immunol., 143(ii), 3666-3672 (1989).
Gamble et al., J. Immunol., Methods, 109, 175-184 (1988).
Poston et al., Am. J. Pathol., 140(3), 665-673 (1992).
Stamper and Woodruff, J. Exp. Med., 144, 828-833 (1976).
Yednock et al., Nature, 356, 63-66 (1992).
Beekhuizen et al., J. Leukocyte Biol., 54, 363-378 (1993).
Bar-Shavit et al., J. Cell. Biol., 112(2), 335-344 (1991).
Allen M. Gown, et al., Human Atherosclerosis II. Immunocytochemical Analysis of the Cellular Composition of Human Atherosclerotic Lesions, Am. J. Pathol. Oct. 1986, 125, 191-207.
Yong-jian Geng et al., Interferon-gamma Inhibits Scavenger Receptor Expression and Foam Cell Formation in Human Monocyte-derived Macrophages, J. Clin. Invest. Apr. 1992, 89, 1322-1330.
Henry Beekhuizen et al., CD14 Contributes to the Adherence of Human Monocytes to Cytokine-Stimulated Endothelial Cells, J. Immunol. Dec. 1, 1991, 147, 3761-3767.
Henry Beekhuizen et al., Characterization of Monocyte Adherence to Human Macrovascular and Microvascular Endothelial Cells, J. Immunol. Jul. 15, 1990, 145, 510-518.
Allard C. van der Wal et al., Adhesion Molecules on the Endothelium and Mononuclear Cells in Human Atherosclerotic Lesions 1992, 141, 1427-1433. Dec. 1992.
Hultquist Steven J.
Leary Louise
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