NDT Advance Access published online on November 27, 2008
Nephrology Dialysis Transplantation, doi:10.1093/ndt/gfn650
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Quantification of circulating endothelial cells in peripheral blood of systemic lupus erythaematosus patients: a simple and reproducible method of assessing endothelial injury and repair
1 Department of Molecular Biology, Genetic Engineering and Biotechnology Institute, Minoufiya University 2 Department of Dermatology, Venereology and Andrology 3 Gastroenterology Surgery Centre, Biotechnology Research-laboratories 4 Urology and Nephrology Center 5 Department of Rheumatology and Rehabilitation, Mansoura University, Egypt 6 Flow Cytometry Core and Hematology Branch, Bethesda, MD, USA
Correspondence and offprint requests to: Khaled Mahmoud, Urology and Nephrology Center, Mansoura University, Egypt. Tel: +2-050-2262222; Fax: +2-050-2263717; E-mail: khaledmahmoud2000{at}hotmail.com
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Abstract |
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Background. Quantification of circulating endothelial cells (CECs) in peripheral blood is developing as a novel and reproducible method of assessing endothelial damage/dysfunction. Accordingly, elevated levels of CECs may be a marker of vascular injury in systemic lupus erythaematosus (SLE). This study was undertaken to assess the blood level of CECs in SLE and to correlate its level with the activity of the disease and to find out the possibility that the presence of increased numbers of CECs can be used as a marker of immune-mediated vessel damage.
Methods. The study included 33 patients with SLE and 20 healthy controls. They were subjected to clinical examination together with laboratory investigations including complete blood count (CBC), erythrocyte sedimentation rate (ESR), urine analysis, renal function test, C3, C4, ANA, anti-ds DNA antibody, antiphospholipid (IgM and IgG) antibodies and quantification of CECs in blood. CECs were calculated using flow cytometry after staining with a mouse anti-human CD45 antibody (pan-leukocyte marker), mouse anti-human CD146 antibody (endothelial cell marker) and 7-amino-actinomycin D (7-AAD) viability marker. CECs were defined as the live cells with 7-AAD negative, CD45 negative and CD146 positive.
Results. The number of CECs was significantly higher in patients with SLE compared with those in healthy control (mean ± SD 38.6 ± 21.2 versus 7.4 ± 3.4). Furthermore, CECs were correlated positively with SLE disease activity index (SLEDAI) score, ESR and anti-ds DNA. CECs from patients with vasculitic skin lesions, renal and central nervous system (CNS) manifestation were significantly higher than patients free from the previous signs.
Conclusions. An increased number of CECs observed in patients with SLE was associated with the active phase of the disease and may represent a marker of widespread endothelial injury.
Keywords: CD146; flow cytometry; systemic lupus erythaematosus
Received for publication: 4. 3.08
Accepted in revised form: 29.10.08