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

doi:10.1016/S0966-3274(99)80008-2

Transplant Immunology

Volume 7, Issue 4, December 1999, Pages 239-246

https://doi.org/10.1016/S0966-3274(99)80008-2 Get rights and content

Abstract

The purpose of this study was to assess the suitability of using endothelial cell (EC) lines for studies of endothelial/immune interactions. The immortal human EC lines HMEC-1, ECV304 and EaHy926 were compared to human umbilical vein endothelial cells (HUVEC) for constitutive and induced expression of surface antigens known to be involved in interactions with T cells. These cell lines were also compared to HUVEC in transendothelial migration assays. Flow cytometry was used to measure cell surface expression of platelet/endothelial cell adhesion molecule-1 (PECAM-1), intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), E-selectin, major histocompatibility complex (MHC) class I and MHC class II, CD40, CD95 (fas) and lymphocyte function associated antigen-3 (LFA-3) before and after treatment with the cytokines tumour necrosis factor-α (TNF-α) and interferon-γ (IFN-γ). Polymerase chain reaction (PCR) was used to detect expression of the MHC class II transactivator.

Significant differences were found in the ability to respond to cytokines between HUVEC and the cell lines, the greatest differences being induction of VCAM-1 and E-selectin in response to TNF-α and induction of MHC class II antigens in response to IFN-γ. Thus unlike HUVEC, induction of VCAM-1 and E-selectin was not detectable on EaHy926 and ECV304 and barely detectable on HMEC-1. MHC class II antigens were not induced on ECV304 in response to IFN-γ and nor was the class II transactivator (CIITA). Unlike HUVEC and the other cell lines, ECV304 were constitutively negative for PECAM-1. Constitutive and induced expression of MHC class I, ICAM-1, LFA/3, CD40 and fas were most conserved between the cell lines and showed little difference to HUVEC. The migration of peripheral blood mononuclear cells (PBMC) through all cell lines was significantly reduced compared to through HUVEC, suggesting that there is a functional difference between the cell lines with regard to interactions with lymphocytes. In conclusion this study has demonstrated significant differences in the ability of endothelial cell lines to respond to cytokines compared to primary HUVEC cultures. In particular ECV304 compares very poorly with HUVEC. Whether these differences are caused by immortalization procedures or reflect heterogeneity of EC arising from different vascular beds is discussed.

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

Abstract

J Invest Dermatol

Cell

J Biol Chem

Cell

J Invest Dermatol

Exp Cell Res

Permanent cell line expressing human factor VIII-related antigen established by hybridization

Proc Natl Acad Sci USA

Spontaneous transformation and immortalization of human endothelial cells in vitro

Cell Dev Biol

Can graft endothelial cells initiate a host anti-graft immune response?

Transplantation

Human endothelial cells effectively costimulate cytokine production by, but not differentiation of, naive CD4+ T cells

J Immunol

Human vasacular endothelial cells process and present autoantigen to human T cell lines

Int Immunol

Induction of ICAM-1 by TNF-alpha, IL-1 beta, and LPS in human endothelial cells after downregulation of PKC

Am J Physiol

IL-4 decreases IFN-gamma-induced endothelial ICAM-1 expression by a transcriptional mechanism

Scand J Immunol

Ligation of ICAM-1 on endothelial cells lead to expression of VCAM-1 via an NF-kB independent mechanism

J Immunol

Activation and homologous desensitization of human endothelial cells by CD40 ligand, tumor necrosis factor, and interleukin 1

J Exp Med

A novel role for E- and P-selectins: shape control of endothelial cell monolayers

J Cell Sci