PERSPECTIVES OF APPLICATION OF ANTIBODIES AGAINST ENDOGLIN (CD105) FOR VISUALIZATION AND ANTI-ANGIOGENE THERAPY OF TUMORS

During last years monoclonal antibodies (MAB) directed against vascular endothelium markers demonstrated their efficiency for visualization and targeted delivery of therapeutic drugs to tumors. Endoglin (CD105) which serves as a key element that determines endothelial cells quiescence or activation is one of such markers. Endoglin is highly expressed on the vascular endothelium of growing tumors. A first panel of MAB against endoglin in our country was produced at the hybridoma technology laboratory of RRC RST named after A.M. Granov. On the basis of these MAB ELISA was created allowing detection of endoglin in human plasma and other biological fluids. Several MAB had been shown to bind endoglin on the membrane of the cultured endothelial cells and to persist there for several hours. During the first 30 min after binding some of the immune complexes “endoglin-MAB” were internalized into the cytoplasm and were found included in the endosomes. In future these MAB can be used to create the reagents for the addressed delivery of isotope tags both on the membrane and into the cytoplasm of endothelial cells.

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Determination of immunoreactive endothelin in medium from cultured endothelial cells and human plasma

Biochemical and Biophysical Research Communications ◽

10.1016/0006-291x(89)91721-x ◽

1989 ◽

Vol 164 (1) ◽

pp. 326-332 ◽

Cited By ~ 26

Author(s):

Yu-Ting Xuan ◽

A.R. Whorton ◽

E. Shearer-Poor ◽

J. Boyd ◽

W.D. Watkins

Keyword(s):

Endothelial Cells ◽

Human Plasma ◽

Cultured Endothelial Cells

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Synthesis of fibronectin by cultured human endothelial cells.

Journal of Experimental Medicine ◽

10.1084/jem.147.6.1779 ◽

1978 ◽

Vol 147 (6) ◽

pp. 1779-1791 ◽

Cited By ~ 311

Author(s):

E A Jaffe ◽

D F Mosher

Keyword(s):

Extracellular Matrix ◽

Endothelial Cells ◽

Human Plasma ◽

Culture Medium ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Basement Membranes ◽

Plasma Fibronectin ◽

Human Endothelial Cells ◽

Cultured Endothelial Cells

Plasma fibronectin is probably the major nonimmune particulate opsonin in blood and is cross-linked to fibrin during the final stage of blood coagulation. Fibronectin also occurs in an insoluble form in basement membranes especially those underlying endothelial cells and in loose connective tissue. Fibronectin was demonstrated in cultured human endothelial cells and in the surrounding extracellular matrix by immunofluorescence microscopy by using antibody to human plasma fibronectin. Cultured human endothelial cells released fibronectin into the culture medium which was immunologically identical to the fibronectin in human plasma. Cultured human endothelial cells were labeled with [3H] leucine. The radioactive fibronectin present in the endothelial postculture medium and in urea extracts of cellular monolayers was isolated with either anti-fibronectin coupled to Protein A-Sepharose or double antibody immunoprecipitation and characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. When reduced, the [3H] fibronectin synthesized by cultured endothelial cells had the same mol wt (approximately 200,000) as plasma fibronectin. Unreduced, the [3H] fibronectin synthesized by endothelial cells migrated as a dimer, as did plasma fibronectin. Fibronectin accounted for approximately 15% of the protein synthesized and released by endothelial cells into the culture medium. Thus, cultured endothelial cells synthesize fibronectin, secrete it into the culture medium, and incorporate it into extracellular matrix. The results suggest that the endothelial cell is potentially a major site of synthesis of circulating plasma fibronectin. In addition, fibronectin derived from endothelial cells may be an important structural component of the subendothelium.

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The Presence in Normal Plasma, Serum and Platelets of Factors That Stimulate the Production of Prostacyclin (PGI2) by Cultured Endothelial Cells

Clinical Science ◽

10.1042/cs0640387 ◽

1983 ◽

Vol 64 (4) ◽

pp. 387-394 ◽

Cited By ~ 24

Author(s):

J. M. Seid ◽

P. B. B. Jones ◽

R. G. G. Russell

Keyword(s):

Endothelial Cells ◽

Vascular Endothelium ◽

Normal Subjects ◽

Disease States ◽

Normal Plasma ◽

Cultured Endothelial Cells ◽

Porcine Endothelial Cells ◽

Dose Dependent ◽

Prostacyclin Production

1. The effect of plasma and serum from normal subjects on the production of prostacyclin by cultured porcine endothelial cells was investigated. 2. Both plasma and serum from all subjects studied significantly stimulated the production of prostacyclin by cultured endothelial cells, measured by the radioimmunoassay of its stable metabolite 6-oxoprostaglandin F1α. 3. Serum caused a consistently greater stimulation than plasma from the same individual. The stimulation was dose-dependent and inhibited by indomethacin. Heparin added to serum also inhibited this response. 4. Extracts from isolated washed platelets were tested for their ability to increase prostacyclin production. Extracts from platelets which had been induced to aggregate and release their granule contents in response to thrombin, caused stimulation. 5. These results indicated the invariable presence in plasma and serum of factors that stimulate the production of prostacyclin by endothelial cells in vitro. At least one of these factors is derived from platelets. These factors may be involved in the regulation of prostacyclin production by the vascular endothelium under normal conditions and in disease states.

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Effect of human plasma lipoproteins on prostacyclin production by cultured endothelial cells.

The Journal of Lipid Research ◽

10.1016/s0022-2275(20)34371-6 ◽

1985 ◽

Vol 26 (3) ◽

pp. 288-297 ◽

Cited By ~ 1

Author(s):

A A Spector ◽

A M Scanu ◽

T L Kaduce ◽

P H Figard ◽

G M Fless ◽

...

Keyword(s):

Endothelial Cells ◽

Human Plasma ◽

Plasma Lipoproteins ◽

Cultured Endothelial Cells ◽

Prostacyclin Production

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Th-P15:20 Thiazolidinediones inhibit endothelial lipase expression in cultured endothelial cells and human plasma

Atherosclerosis Supplements ◽

10.1016/s1567-5688(06)81980-x ◽

2006 ◽

Vol 7 (3) ◽

pp. 496

Author(s):

K. Badellino ◽

K. Trindade ◽

P. Szapary ◽

D.J. Rader

Keyword(s):

Endothelial Cells ◽

Human Plasma ◽

Endothelial Lipase ◽

Cultured Endothelial Cells ◽

Lipase Expression

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The hybridoma-an immunochemical laser.

Clinical Chemistry ◽

10.1093/clinchem/27.9.1580 ◽

1981 ◽

Vol 27 (9) ◽

pp. 1580-1585 ◽

Cited By ~ 24

Author(s):

G S David ◽

R Wang ◽

R Bartholomew ◽

E D Sevier ◽

T H Adams ◽

...

Keyword(s):

Monoclonal Antibodies ◽

Electrophoretic Mobility ◽

Calibration Curve ◽

Immune Complexes ◽

The Other ◽

Immunometric Assay ◽

Specific Affinity ◽

Hybridoma Technology ◽

Antibody Reaction ◽

Antigen Antibody

Abstract The advent of hybridoma technology has provided the immunotechnologist a finely tunable instrument that should permit a marked advance in the immunologic sciences. The ability to choose the precise antibody required and the virtually unlimited availability of easily purified antibodies have already resulted in the simultaneous immunometric assay and potential reagents for immunoscintigraphy and immunotherapy. Using an antibody with a specific affinity and recognition for a specific antigenc determinant can greatly influence the shape and range of a radioimmunometric calibration curve. With monoclonal antibodies, assay systems based on immune complexes (e.g., turbidimetric assays and counterimmunoelectrophoresis) can be made more precise, thus permitting study of the basic physicochemical principles underlying the antigen-antibody reaction and the development of greatly improved quantitative assays. On the other hand, the ability to select an antibody exhibiting specific characteristics implies the necessity to select. One no longer has the luxury of using a mixture of antibodies, hoping to take advantage of the fact that some will have desirable secondary characteristics such as electrophoretic mobility or the ability to absorb to plastics.

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Trophic Effects of Platelets on Cultured Endothelial Cells are Mediated by Platelet-associated Fibroblast Growth Factor-2 (FGF-2) and Vascular Endothelial Growth Factor (VEGF)

Thrombosis and Haemostasis ◽

10.1055/s-0037-1613311 ◽

2002 ◽

Vol 88 (11) ◽

pp. 834-842 ◽

Cited By ~ 68

Author(s):

Giuseppe Pintucci ◽

Scott Froum ◽

Jared Pinnell ◽

Paolo Mignatti ◽

Shahin Rafii ◽

...

Keyword(s):

Endothelial Cells ◽

Growth Factors ◽

Endothelial Cell ◽

Growth Factor ◽

Cell Survival ◽

Vascular Endothelium ◽

Endothelial Cell Proliferation ◽

Nuclear Staining ◽

Endothelial Cell Survival ◽

Cultured Endothelial Cells

SummaryIn addition to their role in primary hemostasis, platelets serve to support and maintain the vascular endothelium. Platelets contain numerous growth factors including the potent angiogenic inducers VEGF and FGF-2. To characterize the function of these two plateletassociated growth factors, the effects of the addition of purified platelets to cultured endothelial cells were examined. The survival and proliferation of endothelial cells were markedly stimulated (2-3-fold and 5-15-fold respectively) by the addition of gel-filtered platelets. Acetylsalicylic acid-treated or lyophilized fixed-platelets were ineffective in supporting endothelial cell proliferation. In Transwell assays, the stimulatory effect of platelets on endothelial cells was preserved, consistent with an effect mediated by secreted factors. The combined inhibition of VEGF and FGF-2 by neutralizing antibodies, in contrast to inhibition of either alone, abrogated both platelet-induced endothelial cell survival and proliferation. FGF-2 isoforms were detected in platelet lysates, as well as in the releasates of agonist-stimulated platelets. Megakaryocytes generated by ex vivo expansion of hematopoietic progenitor cells with kit ligand and thrombopoietin were analyzed for expression of FGF-2. Punctate cytoplasmic staining but no nuclear staining was observed by immunocytochemistry consistent with possible localization of the growth factor to cytoplasmic granules. The addition of platelets to cultured endothelial cells activated extracellular signal-regulated kinase (ERK) in a dose and time-dependent manner. This effect was abrogated by both anti-FGF-2 and anti-VEGF antibody. Since FGF-2 and VEGF are potent angiogenic factors and known endothelial cell survival factors, their release by platelets provides a plausible mechanism for the platelet support of vascular endothelium.

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Human Prostaglandin Transporter Gene ( hPGT ) is Regulated by Fluid Mechanical Stimuli in Cultured Endothelial Cells and Expressed in Vascular Endothelium in Vivo

Circulation ◽

10.1161/01.cir.98.22.2396 ◽

1998 ◽

Vol 98 (22) ◽

pp. 2396-2403 ◽

Cited By ~ 37

Author(s):

James N. Topper ◽

Jiexing Cai ◽

George Stavrakis ◽

Keith R. Anderson ◽

Elizabeth A. Woolf ◽

...

Keyword(s):

Endothelial Cells ◽

Vascular Endothelium ◽

Mechanical Stimuli ◽

Transporter Gene ◽

Cultured Endothelial Cells

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Cross-Linked αsγt-Chain Hybrids in Plasma Clots Studied by 1D- and 2D Electrophoresis and Western Blotting

Thrombosis and Haemostasis ◽

10.1055/s-0038-1649601 ◽

1993 ◽

Vol 70 (03) ◽

pp. 438-442 ◽

Cited By ~ 6

Author(s):

B Grøn ◽

C Filion-Myklebust ◽

S Bjørnsen ◽

P Haidaris ◽

F Brosstad

Keyword(s):

Molecular Weight ◽

Monoclonal Antibodies ◽

Human Plasma ◽

Western Blotting ◽

Isoelectric Point ◽

Factor Xiii ◽

Cross Linking ◽

2D Electrophoresis ◽

Complex Phenomenon ◽

Fibrinopeptide A

SummaryFibrinogen and fibrin related chains in reduced human plasma as well as the bonds interlinking partially cross-linked fibrin from plasma clots have been studied by means of 1D- and 2D electrophoresis and Western blotting. Immunovisualization of reduced plasma or partially cross-linked fibrin with monoclonal antibodies specific for the α-chains or the γ-chains have shown that several bands represent material belonging to both chains. In order to decide whether these bands constitute αγ-chain hybrids or superimposed α- and γ-chain dimers, the cross-linked material was separated according to both isoelectric point (pI) and molecular weight (MW) using Pharmacia’s Multiphor II system. Western blotting of the second dimension gels revealed that partially cross-linked fibrin contains αsγt-chain hybrids and γ- polymers, in addition to the well-known γ-dimers and α-polymers. The main αsγt-chain hybrid has a pI between that of the α- and the γ-chains, a MW of about 200 kDa and contains Aα-chains with intact fibrinopeptide A (FPA). It was also observed that soluble fibrinogen/fibrin complexes as well as partially cross-linked fibrin contain degraded α-dimers with MWs close to the γ-dimers. These findings demonstrate that factor XIII-catalyzed cross-linking of fibrin is a more complex phenomenon than earlier recognized.

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