S‐Endo 1, a pan‐endothelial monoclonal antibody recognizing a novel human endothelial antigen

Bardin, Nathalie; George, F.; Mutin, M; Brisson, Christine; Horschowski, N; Francès, V.; Lesaule, G.; Sampol, José

doi:10.1111/j.1399-0039.1996.tb02666.x

Cited by 127 publications

(91 citation statements)

References 15 publications

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“…CD146 is ubiquitously expressed on endothelial cells [28] and melanoma cells [29]. A correlation with poor prognosis has recently been found in breast cancer; of 635 primary breast tumors, 7% were CD146-positive, and CD146 expression was associated with poor prognosis [12].…”

Section: Discussionmentioning

confidence: 99%

Detection of circulating tumor cells in breast cancer may improve through enrichment with anti-CD146

Mostert

Kraan

Vries

et al. 2010

Breast Cancer Res Treat

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PurposeMost assays to detect circulating tumor cells (CTCs) rely on EpCAM expression on tumor cells.Recently, our group reported that in contrast to other molecular breast cancer subtypes, "normallike" cell lines lack EpCAM expression and are thus missed when CTCs are captured with EpCAM-based technology [1]. Here, the use of CD146 is introduced to detect EpCAM-negative CTCs, thereby improving CTC detection. Methods CD146and EpCAM expression were assessed in our panel of 41 breast cancer cell lines. Cells from 14 cell lines, 9 of which normal-like, were spiked into healthy donor blood. Using CellSearch™ technology, 7.5 mL whole blood was enriched for CTCs by adding ferrofluids loaded with antibodies against EpCAM and/or CD146 followed by staining for Cytokeratin and DAPI. Hematopoietic cells and circulating endothelial cells (CECs) were counterstained with CD45 and CD34, respectively. A similar approach was applied for blood samples of 20 advanced breast cancer patients. ResultsEight of 9 normal-like breast cancer cell lines lacked EpCAM expression but did express CD146.Five of these 8 could be adequately recovered by anti-CD146 ferrofluids. Of 20 advanced breast cancer patients whose CTCs were enumerated with anti-EpCAM and anti-CD146 ferrofluids, 9 had CD146+ CTCs. ConclusionsCells from breast cancer cell lines that lack EpCAM expression frequently express CD146 and can be recovered by anti-CD146 ferrofluids. CD146+ CTCs are present in the peripheral blood of breast cancer patients with advanced disease. Combined use of anti-CD146 and anti-EpCAM is likely to improve CTC detection in breast cancer patients.3

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Section: Discussionmentioning

confidence: 99%

Detection of circulating tumor cells in breast cancer may improve through enrichment with anti-CD146

Mostert

Kraan

Vries

et al. 2010

Breast Cancer Res Treat

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“…Sickle cell disease is a vascular disease that causes tissue ischemia and exaggerated endothelial turnover, both of which are characterized by evident endothelial injury and regeneration [34]. In different patient groups, the number of ECs/mL of peripheral blood may vary between zero and 1,500; most studies found 1 to 100 circulating ECs/mL in healthy people [34][35][36][37][38][39][40]. We found much higher numbers of circulating ECs in both sickle cell patients and controls, probably due to methodological differences in sample preparation and cell quantification.…”

Section: Discussionmentioning

confidence: 99%

Flow cytometric evaluation of circulating endothelial cells: A new protocol for identifying endothelial cells at several stages of differentiation

et al. 2007

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Several factors may influence the analysis of endothelial cells (ECs) by flow cytometry: separation of mononuclear cell, washing and centrifugation steps, panel of monoclonal antibodies, and the lack of standardization of gating technique. Therefore, the reliable quantification of ECs remains a technical challenge. The purpose of this study is to define a new flow cytometric protocol to characterize and quantitate ECs. In previous investigations, increased numbers of circulating ECs have been found in sickle cell disease. The patients with sickle cell disease might provide useful material for the study. We performed flow cytometry on whole blood from 20 normal controls and 31 patients with sickle cell disease (20 patients with steadystate disease and 11 patients with vaso-occlusive crises) using a lyse/no-wash procedure, specific and nonspecific antibody combinations (CD146, CD144, CD34, and CD117), and broad gating. This protocol produced much higher values for the number of circulating ECs (a mean of 2,396.55 ± 658.37 ECs/mL in controls vs 6,709.60 ± 1,772.32 ECs/mL in the steady-state group, or 18,213.50 ± 8,451 in the vaso-occlusive crises group, P < 0.001 for both), and also showed variable EC size and granularity, which may reflect activated, or early release ECs. This novel protocol performed comparably in terms of reproducibility, reliability, and dilution linearity with a previously described protocol. This approach has significant advantages for the characterization and quantitation ECs compatible with the pathophysiology. Using the specific antibodies, CD146 and CD144, together may give more informative EC data than the general approach used. Am. J. Hematol. 82:706-711, 2007. V

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“…Typically, 100 μl of bead suspension is then non-covalently coated with 10 μg/ml CD-146 antibody (Novus Biologicals, Littleton, CO, USA). CD-146 is expressed almost exclusively in mature endothelial cells with the exception of some tumour cell lines [23,24].…”

Section: Methodsmentioning

confidence: 99%

Circulating endothelial cells are elevated in patients with type 2 diabetes mellitus independently of HbA1c

et al. 2005

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Aims/hypothesis: Patients with diabetes mellitus are well known to be at high risk for vascular disease. Circulating endothelial cells (CECs) have been reported to be an ex vivo indicator of vascular injury. We investigated the presence of CECs in the peripheral blood of 25 patients with diabetes mellitus and in nine nondiabetic control donors. Methods: Endothelial cells were isolated from peripheral blood with anti-CD-146-coated immunomagnetic Dynabeads, and were stained with acridine orange dye and counted by fluorescence microscopy. The cells were also stained for von Willebrand factor and Ulex europaeus lectin 1. Results: Patients with diabetes mellitus had an elevated number of CECs (mean 69±30 cells/ml, range 35-126) compared with healthy controls (mean 10±5 cells/ml, range 3-18) (p<0.001). The increase in CECs did not correlate with the levels of HbA 1 c. Circulating endothelial cell numbers were elevated regardless of glucose levels, suggesting that, even with control of glucose levels, there is increased endothelial cell sloughing. Conclusions: Our study suggests that the higher number of CECs in patients with type 2 diabetes may reflect ongoing vascular injury that is not directly dependent on glucose control.

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S‐Endo 1, a pan‐endothelial monoclonal antibody recognizing a novel human endothelial antigen

Cited by 127 publications

References 15 publications

Detection of circulating tumor cells in breast cancer may improve through enrichment with anti-CD146

Detection of circulating tumor cells in breast cancer may improve through enrichment with anti-CD146

Flow cytometric evaluation of circulating endothelial cells: A new protocol for identifying endothelial cells at several stages of differentiation

Circulating endothelial cells are elevated in patients with type 2 diabetes mellitus independently of HbA1c

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