Heterogeneity of Vascular Endothelial Cells: Differences in Susceptibility to Neutrophil-mediated Injury

Murphy, Hedwig S.; Bakopoulos, Natalie; Dame, Michael K.; Varani, James; Ward, Peter A.

doi:10.1006/mvre.1998.2110

Cited by 57 publications

(41 citation statements)

References 16 publications

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“…Endothelial cells were isolated from 3-wk-old C57BL/6 mice. PVEC were isolated from peripheral lung as previously described (5,26). Briefly, strips of peripheral lung were removed, minced, and incubated in 1% gelatincoated 25-cm 2 tissue culture flasks in growth media.…”

Section: Methodsmentioning

confidence: 99%

Disparate mechanisms of sICAM-1 production in the peripheral lung: contrast between alveolar epithelial cells and pulmonary microvascular endothelial cells

Mendez

Morris²,

Wilcoxen³

et al. 2008

American Journal of Physiology-Lung Cellular and Molecular Phys

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Mendez MP, Morris SB, Wilcoxen S, Du M, Monroy YK, Remmer H, Murphy H, Christensen PJ, Paine III R. Disparate mechanisms of sICAM-1 production in the peripheral lung: contrast between alveolar epithelial cells and pulmonary microvascular endothelial cells. Am J Physiol Lung Cell Mol Physiol 294: L807-L814, 2008. First published February 15, 2008 doi:10.1152/ajplung.00398.2007.-Membrane-associated intercellular adhesion molecule-1 (mICAM-1; CD54) is constitutively expressed on the surface of type I alveolar epithelial cells (AEC). Soluble ICAM-1 (sICAM-1) may be produced by proteolytic cleavage of mICAM-1 or by alternative splicing of ICAM-1 mRNA. In contrast to inducible expression seen in most cell types, sICAM-1 is constitutively released by type I AEC and is present in normal alveolar lining fluid. Therefore, we compared the mechanism of sICAM-1 production in primary cultures of two closely juxtaposed cells in the alveolar wall, AEC and pulmonary microvascular endothelial cells (PVEC). AEC, but not PVEC, demonstrated high-level baseline expression of sICAM-1. Stimulation of AEC with TNF␣ or LPS resulted in minimal increase in AEC sICAM-1, whereas PVEC sICAM-1 was briskly induced in response to these signals. AEC sICAM-1 shedding was significantly reduced by treatment with a serine protease inhibitor, but not by cysteine, metalloprotease, or aspartic protease inhibitors. In contrast, none of these inhibitors effected sICAM-1 expression in PVEC. RT-PCR, followed by gel analysis of total RNA, suggests that alternatively spliced fragments are present in both cell types. However, a 16-mer oligopeptide corresponding to the juxtamembrane region of mICAM-1 completely abrogated sICAM-1 shedding in AEC but reduced stimulated PVEC sICAM-1 release by only 20%. Based on these data, we conclude that the predominant mechanism of sICAM-1 production likely differs in the two cell types from opposite sides of the alveolar wall.regulation; mouse; cell culture; CD54 INTERCELLULAR ADHESION MOLECULE-1 (ICAM-1) is an ϳ100-kDa molecule belonging to the immunoglobulin supergene family. The membrane-bound form of this protein (mICAM-1) serves as a counter receptor for the ␤ 2 -integrins, CD11a/CD18 (LFA-1) and CD11b/CD18 (Mac-1), found on leukocytes. Interactions with mICAM-1 facilitate leukocyte transmigration across the endothelium (34) and over the surface of alveolar epithelial cells (AEC) in the lung (27). A soluble form of the molecule, soluble intercellular adhesion molecule-1 (sICAM-1), is found in serum and in the alveolar lining fluid (7,19,23). sICAM-1 may be generated by proteolytic cleavage and/or alternative splicing of mICAM-1 messenger RNA (4, 35, 37). Like mICAM-1, sICAM-1 interacts with LFA-1/Mac-1 to compete with leukocyte binding to mICAM-1 (36) and to stimulate leukocytes (31).sICAM-1 is normally present in the alveolar lining fluid of humans and mice (7,13,14,16,17,23). We have previously demonstrated that type I AEC are the likely source of sICAM-1 in the alveolar lining fluid and that sICAM-1 is constitutively...

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Section: Methodsmentioning

confidence: 99%

Disparate mechanisms of sICAM-1 production in the peripheral lung: contrast between alveolar epithelial cells and pulmonary microvascular endothelial cells

Mendez

Morris²,

Wilcoxen³

et al. 2008

American Journal of Physiology-Lung Cellular and Molecular Phys

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“…Ears were removed, split, and incubated in dispase II (5 mg/ml in PBS) for 45 min, after which the epidermis was removed and discarded. Endothelial cells were expressed from the dermal sheets and placed into growth medium (RPMI medium supplemented with endothelial cell growth factor, 20% heat-inactivated FCS, L-glutamine, streptomycin-penicillin, and fungizone) in gelatin-coated tissue culture dishes by applying lateral pressure with the blunt end of a scalpel according to a previously described method (41). Cells grown to confluence in 4 -6 days were trypsinized and used at 90 -95% confluence for all experimental studies (passage 1).…”

Section: Mdmec Culturementioning

confidence: 99%

Expression and Function of C5a Receptor in Mouse Microvascular Endothelial Cells

Laudes¹,

Chu²,

Huber‐Lang³

et al. 2002

The Journal of Immunology

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150

110

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The complement-derived anaphylatoxin, C5a, is a potent phlogistic molecule that mediates its effects by binding to C5a receptor (C5aR; CD88). We now demonstrate specific binding of radiolabeled recombinant mouse C5a to mouse dermal microvascular endothelial cells (MDMEC) with a Kd50 of 3.6 nM and to ∼15,000–20,000 receptors/cell. Recombinant mC5a competed effectively with binding of [125I]rmC5a to MDMEC. Enhanced binding of C5a occurred, as well as increased mRNA for C5aR, after in vitro exposure of MDMEC to LPS, IFN-γ, or IL-6 in a time- and dose-dependent manner. By confocal microscopy, C5aR could be detected on surfaces of MDMEC using anti-C5aR Ab. In vitro expression of macrophage inflammatory protein-2 (MIP-2) and monocyte chemoattractant protein-1 (MCP-1) by MDMEC was also measured. Exposure of MDMEC to C5a or IL-6 did not result in changes in MIP-2 or MCP-1 production, but initial exposure of MDMEC to IL-6, followed by exposure to C5a, resulted in significantly enhanced production of MIP-2 and MCP-1 (but not TNF-α and MIP-1α). Although LPS or IFN-γ alone induced some release of MCP-1 and MIP-2, pre-exposure of these monolayers to LPS or IFN-γ, followed by addition of C5a, resulted in synergistic production of MIP-2 and MCP-1. Following i.v. infusion of LPS into mice, up-regulation of C5aR occurred in the capillary endothelium of mouse lung, as determined by immunostaining. These results support the hypothesis that C5aR expression on MDMEC and on the microvascular endothelium of lung can be up-regulated, suggesting that C5a in the co-presence of additional agonists may mediate pro-inflammatory effects of endothelial cells.

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“…All cells used for functional or cytofluorographic analysis were studied 6 days after challenge. Dermal microvascular endothelial cells from AKR mice were isolated according to a previously published protocol (21). Identity of the isolated endothelial cells was confirmed by their typical cobblestone appearance and Von Willibrand factor (21) and CD34 (22) expression.…”

Section: Mice and Peritoneal M Isolationmentioning

confidence: 99%

CD49d Overexpression and T Cell Autoimmunity

Eisenbraun

Sonstein

et al. 2003

The Journal of Immunology

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D10.G4.1 (D10) cells, a murine conalbumin-reactive Th2 cell line, made to overexpress the β2 integrin LFA-1 by pharmacological manipulation or by transfection become autoreactive and are capable of inducing in vivo autoimmunity. However, whether this is specific to LFA-1 and whether overexpression of other T cell integrin molecules has the same effect are unknown. We examined the functional consequences of T cell CD49d (α4 integrin) overexpression by transfecting murine CD49d cDNA into D10 cells. Similar to the LFA-1-transfected cells, the CD49d-overexpressing T cells are autoreactive and proliferate in response to APCs in an MHC class II-dependent manner in the absence of nominal Ag. Additionally, CD49d overexpression is associated with increased in vitro adhesion to endothelial cells and increased in vivo splenic homing. However, in contrast to LFA-1 overexpression, increased T cell CD49d expression is not associated with autoreactive cytotoxicity or the ability to induce in vivo autoimmunity. In addition to the novel observation that CD49d overexpression is sufficient to induce T cell autoreactivity, our results also support the hypothesis that the ability to induce in vivo autoimmunity is related to T cell cytotoxicity and not to T cell proliferation function in the D10 murine adoptive transfer model of autoimmunity.

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Heterogeneity of Vascular Endothelial Cells: Differences in Susceptibility to Neutrophil-mediated Injury

Cited by 57 publications

References 16 publications

Disparate mechanisms of sICAM-1 production in the peripheral lung: contrast between alveolar epithelial cells and pulmonary microvascular endothelial cells

Disparate mechanisms of sICAM-1 production in the peripheral lung: contrast between alveolar epithelial cells and pulmonary microvascular endothelial cells

Expression and Function of C5a Receptor in Mouse Microvascular Endothelial Cells

CD49d Overexpression and T Cell Autoimmunity

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