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首页> 外文期刊>Transplant immunology >A comparison of primary endothelial cells and endothelial cell lines for studies of immune interactions.
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A comparison of primary endothelial cells and endothelial cell lines for studies of immune interactions.

机译：比较原代内皮细胞和内皮细胞系以研究免疫相互作用。

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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-alpha (TNF-alpha) and interferon-gamma (IFN-gamma). 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-alpha and induction of MHC class II antigens in response to IFN-gamma. 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-gamma 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.
机译：这项研究的目的是评估使用内皮细胞（EC）系进行内皮/免疫相互作用研究的适用性。将永生的人类EC系HMEC-1，ECV304和EaHy926与人类脐静脉内皮细胞（HUVEC）进行了已知与T细胞相互作用涉及的表面抗原的组成型和诱导型表达的比较。这些细胞系也通过内皮迁移试验与HUVEC进行了比较。流式细胞仪用于检测血小板/内皮细胞粘附分子-1（PECAM-1），细胞间粘附分子-1（ICAM-1），血管细胞粘附分子-1（VCAM-1），E-选择素的细胞表面表达，用细胞因子肿瘤坏死因子-α（TNF-alpha）治疗之前和之后的主要组织相容性复合物（MHC）I类和MHC II类，CD40，CD95（fas）和淋巴细胞功能相关抗原3（LFA-3）干扰素-γ（IFN-γ）。聚合酶链反应（PCR）用于检测MHC II类反式激活因子的表达。发现在HUVEC和细胞系之间对细胞因子的反应能力上存在显着差异，最大的差异是对TNF-α的应答诱导VCAM-1和E-选择素和对IFN-γ的应答MHC II类抗原。因此，与HUVEC不同，在EaHy926和ECV304上检测不到VCAM-1和E-选择素的诱导，而在HMEC-1上几乎检测不到。 MHC II类抗原未响应IFN-γ在ECV304上诱导，II类反式激活因子（CIITA）也未诱导。与HUVEC和其他细胞系不同，ECV304对PECAM-1构成性阴性。 MHC I类，ICAM-1，LFA / 3，CD40和fas的组成型和诱导表达在细胞系之间最保守，与HUVEC的表达差异很小。与通过HUVEC相比，通过所有细胞系迁移的外周血单核细胞（PBMC）明显减少，这表明细胞系之间在与淋巴细胞相互作用方面存在功能差异。总而言之，该研究表明内皮细胞系对细胞因子的反应能力与原代HUVEC培养物相比有显着差异。特别是ECV304与HUVEC相比非常差。讨论了这些差异是由永生化程序引起的还是反映了不同血管床引起的EC异质性。

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