Abstract | OBJECTIVE: Blood monocytes are known to express endothelial-like genes during co-culture with endothelium. In this study, the time-dependent change in the phenotype pattern of primary blood monocytes after adhering to endothelium is reported using a novel HLA-A2 mistyped co-culture model. METHODS AND RESULTS: Freshly isolated human PBMCs were co-cultured with human umbilical vein endothelial cells or human coronary arterial endothelial cells of converse human leukocyte antigen A2 (HLA-A2) status. This allows the tracking of the PBMC-derived cells by HLA-A2 expression and assessment of their phenotype pattern over time. PBMCs that adhered to the endothelium at the start of the co-culture were predominantly CD11b+ blood monocytes. After 24 to 72 hours in co-culture, the endothelium-adherent monocytes acquired endothelial-like properties including the expression of endothelial nitric oxide synthase, CD105, CD144 and vascular endothelial growth factor receptor 2. The expression of monocyte/macrophage lineage antigens CD14, CD11b and CD36 were down regulated concomitantly. The adherent monocytes did not express CD115 after 1 day of co-culture. By day 6, the monocyte-derived cells expressed vascular cell adhesion molecule 1 in response to tumour necrosis factor alpha. Up to 10% of the PBMCs adhered to the endothelium. These monocyte-derived cells contributed up to 30% of the co-cultured cell layer and this was dose-dependent on the PBMC seeding density. CONCLUSIONS: Human blood monocytes undergo rapid phenotype change to resemble endothelial cells after adhering to endothelium.
|
Authors | Colin Tso, Kerry-Anne Rye, Philip Barter |
Journal | PloS one
(PLoS One)
Vol. 7
Issue 5
Pg. e37091
( 2012)
ISSN: 1932-6203 [Electronic] United States |
PMID | 22615904
(Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
|
Chemical References |
- Antigens, CD
- HLA-A2 Antigen
- Tumor Necrosis Factor-alpha
- Vascular Cell Adhesion Molecule-1
- Nitric Oxide Synthase Type III
- Vascular Endothelial Growth Factor Receptor-2
|
Topics |
- Antigens, CD
(metabolism)
- Cell Adhesion
(physiology)
- Cells, Cultured
- Coculture Techniques
(methods)
- Down-Regulation
- Endothelium, Vascular
(cytology, metabolism, physiology)
- HLA-A2 Antigen
(metabolism)
- Human Umbilical Vein Endothelial Cells
(cytology, metabolism, physiology)
- Humans
- Leukocytes, Mononuclear
(cytology, metabolism, physiology)
- Macrophages
(metabolism, physiology)
- Monocytes
(cytology, metabolism, physiology)
- Nitric Oxide Synthase Type III
(metabolism)
- Phenotype
- Tumor Necrosis Factor-alpha
(metabolism)
- Vascular Cell Adhesion Molecule-1
(metabolism)
- Vascular Endothelial Growth Factor Receptor-2
(metabolism)
|