Detection and Characterization of Rare Circulating Endothelial Cells by Imaging Flow Cytometry.

Circulating endothelial cells (CECs) are angiogenic cells that appear in increased numbers in the peripheral circulation either as a result of vascular injury or in response to angiogenic stimuli. Elevated levels of CECs have been correlated with various disease states, indicating the use of CECs as a biomarker of disease. Flow cytometry is a widely accepted method for detecting and quantitating CECs. Flow cytometry provides statistical information on large numbers of cells but no information on morphological characteristics. Imaging flow cytometry combines traditional flow cytometry and microscopy, providing a streamlined, multiparameter approach to characterize the biological properties and morphology of large numbers of cells, and is particularly amenable for rare event analysis such as CECs. This approach for identifying and characterizing CECs allows the morphological characterization of large numbers of live, nucleated, single CECs, and alleviates the need for prior enrichment.

[1]  R. Hoffman,et al.  Human CD34+ HLA-DR- bone marrow cells contain progenitor cells capable of self-renewal, multilineage differentiation, and long-term in vitro hematopoiesis. , 1991, Blood cells.

[2]  B. Nickoloff The human progenitor cell antigen (CD34) is localized on endothelial cells, dermal dendritic cells, and perifollicular cells in formalin-fixed normal skin, and on proliferating endothelial cells and stromal spindle-shaped cells in Kaposi's sarcoma. , 1991, Archives of dermatology.

[3]  P. Ramani,et al.  Endothelial markers in malignant vascular tumours of the liver: superiority of QB-END/10 over von Willebrand factor and Ulex europaeus agglutinin 1. , 1991, Journal of clinical pathology.

[4]  CD34: structure, biology, and clinical utility. , 1996, Blood.

[5]  E. Wayner,et al.  Circulating activated endothelial cells in sickle cell anemia. , 1997, The New England journal of medicine.

[6]  N. Voelkel,et al.  Circulating endothelial cells in pulmonary hypertension , 2003, Thrombosis and Haemostasis.

[7]  M. Haubitz,et al.  Circulating endothelial cells as markers for ANCA-associated small-vessel vasculitis , 2003, The Lancet.

[8]  E. Voest,et al.  Increased levels of viable circulating endothelial cells are an indicator of progressive disease in cancer patients. , 2004, Annals of oncology : official journal of the European Society for Medical Oncology.

[9]  H. Maecker,et al.  Selecting fluorochrome conjugates for maximum sensitivity , 2004, Cytometry. Part A : the journal of the International Society for Analytical Cytology.

[10]  J. P. McCoy,et al.  CD146 (Mel-CAM), an adhesion marker of endothelial cells, is a novel marker of lymphocyte subset activation in normal peripheral blood. , 2005, Blood.

[11]  J. P. McCoy,et al.  Detection of circulating endothelial cells and endothelial progenitor cells by flow cytometry , 2005, Cytometry. Part B, Clinical cytometry.

[12]  S. Nourshargh,et al.  PECAM-1: a multi-functional molecule in inflammation and vascular biology. , 2007, Arteriosclerosis, thrombosis, and vascular biology.

[13]  M. Roederer,et al.  Optimizing a multicolor immunophenotyping assay. , 2007, Clinics in laboratory medicine.

[14]  J. P. McCoy,et al.  A unique population of effector memory lymphocytes identified by CD146 having a distinct immunophenotypic and genomic profile , 2007, BMC Immunology.

[15]  D. Nixon,et al.  Nine‐color flow cytometry for accurate measurement of T cell subsets and cytokine responses. Part I: Panel design by an empiric approach , 2008, Cytometry. Part A : the journal of the International Society for Analytical Cytology.

[16]  J. P. McCoy,et al.  CD146+ T lymphocytes are increased in both the peripheral circulation and in the synovial effusions of patients with various musculoskeletal diseases and display pro‐inflammatory gene profiles , 2009, Cytometry. Part B, Clinical cytometry.

[17]  J. Privratsky,et al.  PECAM-1: conflicts of interest in inflammation. , 2010, Life sciences.

[18]  J Philip McCoy,et al.  Imaging flow cytometry for morphologic and phenotypic characterization of rare circulating endothelial cells , 2013, Cytometry. Part B, Clinical cytometry.

[19]  J. P. McCoy,et al.  Studying the human immunome: the complexity of comprehensive leukocyte immunophenotyping. , 2014, Current topics in microbiology and immunology.