A comparison of primary endothelial cells and endothelial cell lines for studies of immune interactions

Lidington, EA; Moyes, David L.; McCormack, Ann; Rose, Marie E.

doi:10.1016/s0966-3274(99)80008-2

Cited by 139 publications

(111 citation statements)

References 30 publications

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“…Unlike HUVECs, which are primary human umbilical vein endothelial cells, HMEC-1 is an immortal human EC line. Significant differences have been found between HUVEC and HMEC-1 cell lines [54]. Thus, together with our current study, we conclude that FBW7 is an indispensable regulator of endothelial functions by targeting multiple key factors, including KLF2, Notch, NF1 and HIF1-α, for degradation in a context-dependent manner.…”

Section: Discussionsupporting

confidence: 84%

FBW7 regulates endothelial functions by targeting KLF2 for ubiquitination and degradation

Wang

Liu

et al. 2013

Cell Res

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F-box and WD repeat domain-containing 7 (FBW7), the substrate-binding subunit of E3 ubiquitin ligase SCF FBW7 (a complex of SKP1, cullin-1 and FBW7), plays important roles in various physiological and pathological processes. Although FBW7 is required for vascular development, its function in the endothelium remains to be investigated. In this study, we show that FBW7 is an important regulator of endothelial functions, including angiogenesis, leukocyte adhesion and the endothelial barrier integrity. Using RNA interference, we found that the depletion of FBW7 markedly impairs angiogenesis in vitro and in vivo. We identified the zinc finger transcription factor Krüppel-like factor 2 (KLF2) as a physiological target of FBW7 in endothelial cells. Knockdown of FBW7 expression resulted in the accumulation of endogenous KLF2 protein in endothelial cells. FBW7-mediated KLF2 destruction was shown to depend on the phosphorylation of KLF2 via glycogen synthase kinase-3 (GSK3) at two conserved phosphodegrons. Mutating these phosphodegron motifs abolished the FBW7-mediated degradation and ubiquitination of KLF2. The siRNAmediated knockdown of FBW7 showed that KLF2 is an essential target of FBW7 in the regulation of endothelial functions. Moreover, FBW7-mediated KLF2 degradation was shown to be critical for angiogenesis in teratomas and in zebrafish development. Taken together, our study suggests a role for FBW7 in the processes of endothelial cell migration, angiogenesis, inflammation and barrier integrity, and provides novel insights into the regulation of KLF2 stability in vivo.

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Section: Discussionsupporting

confidence: 84%

FBW7 regulates endothelial functions by targeting KLF2 for ubiquitination and degradation

Wang

Liu

et al. 2013

Cell Res

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“…There are differences in antigen composition and reactions to stimuli between the endothelium from large and small vessels, and between endothelial cells derived from various microvascular endothelial beds (23). The EAhy926 cell line was derived by fusing HUVECs with a permanent human epithelial cell line A549 (16).…”

Section: Discussionmentioning

confidence: 99%

Pre-transplant serum concentrations of anti-endothelial cell antibody in panel reactive antibody negative renal recipients and its impact on acute rejection

Han

Lü²,

Jin³

et al. 2009

Clinical Chemistry and Laboratory Medicine

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Background: Endothelial cell antigens are important targets in acute rejection (AR). Our goal was to measure the serum concentrations of pre-transplant antiendothelial cell antibody (AECA) in panel reactive antibody (PRA) negative recipients and its impact on AR within 6 months following renal transplantation. Methods: We retrospectively examined pre-transplant sera from 392 patients using cellular enzyme linked immunosorbent assay (ELISA) with substrate from a permanent endothelial cell line EAhy926. Equal volumes of serum from 40 healthy volunteers were mixed and used as the negative control. Results: The positive rate of AECA was 15.8%. There were no significant differences with respect to age, gender, original disease, dialysis history, immune suppressive regimen, cytomegalovirus (CMV) antigen positive rate, complement dependent cytotoxicity (CDC) level and soluble CD30 (sCD30) levels between the AECA positive group and AECA negative group. AR rate in the AECA positive group was higher than that in the AECA negative group (35.5% vs. 22.4%, ps0.023). The AECA positive patients had significantly higher rates of acute grade II T-cell mediated rejection (TMR) and acute antibody mediated rejection (AMR) compared with AECA negative patients. The concentrations of sCD30, and AECA were independent risk factors for AR within 6 months; the odds ratios were 7.005 and 2.469, respectively. Conclusions: Positive AECA was an independent risk factor for AR and appeared to correlate with relatively severe rejection subtypes. Clin Chem Lab Med 2009;47:1265-9.

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“…A deeper knowledge of tissue-derived endothelium might help us understand its physiological properties and the pathological processes occurring within a specific organ. It is, in fact, widely accepted that phenotype and function of microvascular endothelial cells (MECs) derived from different vascular beds are heterogeneous [3][4][5][6][7][8], supporting the proposal that tissue-specific vascular beds have specialised functions.…”

Section: Introductionmentioning

confidence: 99%

Primary and immortalised human pancreatic islet endothelial cells: phenotypic and immunological characterisation

et al. 2005

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Aims/hypothesis: Studies on the biology of the microvascular endothelial cells (MECs) that surround and penetrate the pancreatic islets are hampered by difficulties in isolating and culturing large numbers of pure cells. We aimed to morphologically and functionally characterise primary MECs purified and cultured from human islets, and to establish a simian virus 40 (SV40)-immortalised cell line from these primary cultures. Materials and methods: Human islet MECs were extracted and purified using anti-CD105 coated immunomagnetic beads, and endothelial markers and surface molecules analysed by flow cytometric analysis. An immortalised cell line was then established by using a chimeric adeno5/SV40 virus. Results: Islet MECs expressed classic and specific endothelial markers, a high basal level of intercellular adhesion molecule-1, and low levels of E-selectin and TNF (previously known as TNF-α) inducible vascular cell adhesion molecule-1. IFNG (previously known as IFN-γ) induced expression of HLA class II molecules. The immortalised islet MECs expanded rapidly, exhibited increased DNA synthesis, and were passaged approximately 30 times, without signs of senescence. They retained the endothelial characteristics of the parental cells, and behaved as the primary cells in terms of TNF stimulation of expression of adhesion molecules and support of leucocyte adhesion and transmigration. Conclusions/interpretation: The immortalised islet MECs that we have established could effectively represent a substitute for primary counterparts for in vitro studies on the role of the microvasculature in pathophysiological processes involved in type 1 and type 2 diabetes.

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A comparison of primary endothelial cells and endothelial cell lines for studies of immune interactions

Cited by 139 publications

References 30 publications

FBW7 regulates endothelial functions by targeting KLF2 for ubiquitination and degradation

FBW7 regulates endothelial functions by targeting KLF2 for ubiquitination and degradation

Pre-transplant serum concentrations of anti-endothelial cell antibody in panel reactive antibody negative renal recipients and its impact on acute rejection

Primary and immortalised human pancreatic islet endothelial cells: phenotypic and immunological characterisation

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