Isolation, Culture, and Characterisation of MHC Class II-Positive Microvascular Endothelial Cells from the Human Heart

McDouall, Rhoda; Yacoub, Magdi H.; Rose, Marlene L.

doi:10.1006/mvre.1996.0016

Cited by 57 publications

(40 citation statements)

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“…Our results support this: RMEC constitutively express HLA-DR in vivo, whereas endothelial cells from larger vessels do not; lower levels of CD31 on cultured RMEC distinguish them from HUVEC and cultured human lung microvascular endothelial cells; vWF expression is variable within a population of RMEC that otherwise have a uniform phenotype. Cardiac microvascular endothelial cells have a phenotype similar to RMEC with respect to HLA-DR expression in that endothelial cells of small but not large cardiac vessels express HLA-DR; and HLA-DR expression on isolated cardiac microvascular endothelial cells is lost after 2 wk of culture (22,38).…”

Section: Discussionmentioning

confidence: 99%

Normal Human Kidney HLA-DR–Expressing Renal Microvascular Endothelial Cells

Muczynski¹,

Ekle²,

Coder³

et al. 2003

Journal of the American Society of Nephrology

145

107

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Abstract. Human, but not murine, renal peritubular and glomerular capillaries constitutively express class II major histocompatibility (MHC) proteins at high levels in normal human kidney. Expression of class II proteins on renal microvascular endothelial cells (RMEC) makes it available to circulating lymphocytes and imparts a surveillance capacity to RMEC for controlling inflammatory responses. In this report, the coexpression of HLA-DR and the endothelial marker CD31 are used to identify RMEC as a distinct population of cells within a standard renal biopsy using flow cytometry. A three-laser, multicolor flow cytometry analysis using Alexa dyes, developed for characterizing the expression of cell surface antigens, identifies RMEC as a population separate from HLA-DRexpressing leukocytes. HLA-DR RMEC co-express HLA-DP and HLA-DQ. RMEC also express the T cell costimulatory factor CD58 but not CD80, CD86, or CD40. On the basis of high HLA-DR expression, RMEC are isolated for culture using fluorescence-activated cell sorting and magnetic beads. Cultured RMEC require normal basal physiologic concentrations of gamma interferon (␥IFN) to maintain HLA protein expression. This expression is regulated by CIITA, the MHC class II-specific transcription factor. Four tissue-specific promoters have been described for CIITA. In freshly isolated RMEC, RT-PCR and hybridization using specific oligonucleotide probes to CIITA promoter sequences identify only the statinsensitive ␥IFN-induced promoter IV of CIITA. Therefore, the constitutive expression of HLA-DR on RMEC in normal human kidney is located in a position for immune surveillance, depends on basal physiologic concentrations of ␥IFN, and may be amenable to regulation with statins.MHC proteins are of two classes distinguished on the basis of structure and function. Class I molecules, composed of a polymorphic subunit complexed with ␤-2-microglobulin, are found on all nucleated cells and present antigenic peptides to CD8ϩ T lymphocytes; class II molecules, composed of polymorphic ␣ and ␤ chains, are constitutively expressed on a limited number of cell types (dendritic cells, macrophages, B lymphocytes) and present antigenic peptides to CD4 ϩ T lymphocytes. We recently described an unusual expression of MHC class II proteins in normal human kidneys that is not found in murine kidneys (1). The human MHC class II protein HLA-DR is abundantly expressed on peritubular and glomerular capillary endothelial cells but not on endothelial cells of larger blood vessels of normal kidney. Antibodies to HLA-DR and CD31, a protein highly expressed on endothelial cells, co-localize on peritubular and glomerular cells within sections of kidney tissue, indicating capillary endothelial cell location of HLA-DR. HLA-DR has also been identified on rare scattered circulating leukocytes found within the kidney, but over 98% of the DR identified by immunofluorescence microscopy in kidney cortex is located on capillary endothelial cells (1). We refer to these cells co-expressing HLA-DR and CD31 as rena...

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Section: Discussionmentioning

confidence: 99%

Normal Human Kidney HLA-DR–Expressing Renal Microvascular Endothelial Cells

Muczynski¹,

Ekle²,

Coder³

et al. 2003

Journal of the American Society of Nephrology

145

107

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“…In addition to pattern recognition receptors broadly expressed on most endothelium, e.g., proteins of the Toll-like receptor family (43) or receptors of the scavenger cell pathway of low density lipoprotein metabolism (44), DMEC express other scavenger receptors, which have a very restricted expression pattern. For example, DMEC express CD36 (27), which is otherwise found on endothelium of brain and heart only (45,46), and DMEC express CD32 (21), which is otherwise found on endothelium of liver only (17). This combined expression of multiple pattern recognition receptors is to date unique for DMEC and raises important questions about their roles in innate immunity.…”

Section: Discussionmentioning

confidence: 99%

Dermal Microvascular Endothelial Cells Express the 180-kDa Macrophage Mannose Receptor In Situ and In Vitro

Gröger¹,

Holnthoner²,

Maurer

et al. 2000

The Journal of Immunology

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Expression of the 180-kDa mannose receptor (MR) is mainly found on cells of the macrophage lineage. MR mediates the uptake of micro-organisms and host-derived glycoproteins. We demonstrate that endothelium of the human skin in situ and dermal microvascular endothelial cells (DMEC) in vitro expressed MR at both the protein and mRNA levels. In contrast, HUVEC were consistently negative for MR expression. DMEC internalized dextran as well as Escherichia coli by the way of MR into acidic phagosomes, only a few of which fused with CD63- and lysosomal-associated membrane glycoprotein-2-positive lysosomes. This contrasts with the situation in monocyte-derived dendritic cells, where almost all of the MR-Ag complexes reached CD63- and lysosomal-associated membrane glycoprotein-2-positive compartments, indicating differences in the phagolysosomal fusion rate between DMEC and dendritic cells. In conclusion, DMEC express functional MR, a finding that corroborates a role of skin endothelium in Ag capture/clearing.

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“…All EC were positive for HLA class I (W6/32), but large vessel EC from the pulmonary artery and aorta, as well as those of fetal origin (HUVEC and EAhy.926) were negative for HLA-DR. Primary isolates of MEC were positive for HLA-DR expression, but this expression was lost with time in culture, as previously described. 16 CA were also found to loose basal class II expression in culture. Following IFN-g treatment, all EC were induced to express HLA-DR (>95%) (Fig.…”

Section: Purity and Characterization Of Stimulator Cellsmentioning

confidence: 98%

“…16 The pieces of ventricle, from patients with ischaemic heart disease or cardiomyopathy, appeared normal by gross morphology. Briefly, epicardium and endocardium were discarded to exclude contamination by either type of EC or mesothelial cells, and the tissue digested in 1000 U/ml collagenase type II (Sigma, Poole, UK).…”

Section: Isolation Of Microvessel Ecmentioning

confidence: 99%

Alloproliferation of purified CD4⁺ T cells to adult human heart endothelial cells, and study of second‐signal requirements

et al. 1996

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SUMMARYHuman endothelial cells have been shown to be capable of causing direct allostimulation of T cells. However, the majority of immunological studies of human endothelial cells have been performed on cells of fetal origin. Here we use endothelial cells isolated from the adult human heart, both large vessel (coronary artery, pulmonary artery and aorta) and also microvascular. We have examined the ability of all these endothelial cells to cause direct allostimulation of T cells, and show that purified CD4T cells can proliferate in response to adult human heart endothelial cells, the response being dependent on pretreatment of the endothelial cells with interferon-g (IFN-g) and inhibited by anti-HLA-DR monoclonal antibody. The proliferative responses of CD8 T cells to adult but not fetal endothelial cells was inconsistent and weak. Proliferative responses were not blocked by CTLA4-Ig, which inhibits Tcell responses to 'classical' antigen-presenting cells (APC), but > 50% inhibition was achieved with monoclonal antibody to lymphocyte function-associated antigen-3 (LFA-3). These results show that adult human cardiovascular endothelial cells are capable of causing allostimulation of resting CD4 T cells, using a different second signal to classical APC. In view of these findings endothelial cells should be considered as APC following solid organ transplantation.

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Isolation, Culture, and Characterisation of MHC Class II-Positive Microvascular Endothelial Cells from the Human Heart

Cited by 57 publications

References 0 publications

Normal Human Kidney HLA-DR–Expressing Renal Microvascular Endothelial Cells

Normal Human Kidney HLA-DR–Expressing Renal Microvascular Endothelial Cells

Dermal Microvascular Endothelial Cells Express the 180-kDa Macrophage Mannose Receptor In Situ and In Vitro

Alloproliferation of purified CD4⁺ T cells to adult human heart endothelial cells, and study of second‐signal requirements

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Isolation, Culture, and Characterisation of MHC Class II-Positive Microvascular Endothelial Cells from the Human Heart

Cited by 57 publications

References 0 publications

Normal Human Kidney HLA-DR–Expressing Renal Microvascular Endothelial Cells

Normal Human Kidney HLA-DR–Expressing Renal Microvascular Endothelial Cells

Dermal Microvascular Endothelial Cells Express the 180-kDa Macrophage Mannose Receptor In Situ and In Vitro

Alloproliferation of purified CD4+ T cells to adult human heart endothelial cells, and study of second‐signal requirements

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Alloproliferation of purified CD4⁺ T cells to adult human heart endothelial cells, and study of second‐signal requirements