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Updated: Aug 9, 2026
Isolation of Primary Mouse Lung Endothelial Cells
Published on: November 10, 2021
Murine endothelial cell line cells, F-2: interaction with leukocytes and cytokines production
1Department of Immunobiology, Faculty of Medicine, Kyoto University, Japan.
Insights
Murine endothelial cells (F-2) express adhesion molecules. Tumor necrosis factor-alpha (TNF-α) alters intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) expression, impacting T and B cell adhesion.
Area of Science:
- Immunology
- Cell Biology
- Endothelial Cell Biology
Background:
- Endothelial cells play a crucial role in immune cell trafficking.
- Understanding the expression and regulation of adhesion molecules on endothelial cells is vital for studying immune responses.
Purpose of the Study:
- To characterize the expression of adhesion molecules on the murine endothelial cell line F-2.
- To investigate the effects of cytokines, specifically TNF-alpha, IL-1 alpha, and IFN-gamma, on adhesion molecule expression and immune cell adhesion to F-2 cells.
- To determine the role of ICAM-1 and VCAM-1 pathways in T cell, B cell, and granulocyte adhesion.
Main Methods:
- Flow cytometry was used to analyze the expression of MHC class I, FcR II, Mac-1, VCAM-1, and ICAM-1 on F-2 cells.
- Co-culture experiments were performed with F-2 cells and T (Con A blast) and B (LPS blast) cells.
- Inhibition assays using monoclonal antibodies (mAbs) against ICAM-1, LFA-1, VCAM-1, and VLA-4 were conducted.
- Cytokine production (IL-6, GM-CSF, neutrophil chemotactic activity) by F-2 cells was measured.
Main Results:
- F-2 cells constitutively expressed MHC class I, FcR II, Mac-1, and VCAM-1, but not ICAM-1 or MHC class II.
- TNF-alpha treatment increased ICAM-1 and decreased VCAM-1 expression, while IL-1 alpha and IFN-gamma affected VCAM-1 but not ICAM-1.
- T and B cell adhesion to F-2 cells was enhanced by TNF-alpha, with ICAM-1 preferentially used by T cells and VCAM-1 by B cells.
- Granulocyte adhesion was observed but not inhibited by the tested mAbs.
- TNF-alpha significantly increased IL-6 and neutrophil chemotactic activity production by F-2 cells.
Conclusions:
- Murine endothelial F-2 cells exhibit differential regulation of adhesion molecules by cytokines.
- Cytokine-induced changes in ICAM-1 and VCAM-1 expression modulate T and B cell adhesion pathways.
- F-2 cells contribute to the inflammatory response through the production of cytokines and chemokines.
Abstract:
Flowcytometry demonstrated that murine endothelial cell line F-2 expresses MHC class I antigen, FcR II, Mac-1 and vascular cell adhesion molecule-1 (VCAM-1), but not intercellular adhesion molecule-1 (ICAM-1) and class II antigen. However, co-culturing with TNF-alpha for 24 hr resulted in the increased expression of ICAM-1, and the decreased expression of VCAM-1. IL-1 alpha and IFN-gamma exerted this regulatory effect on VCAM-1 but not on ICAM-1. T (Con A blast) and B (LPS blast) cells adhered to F-2 cells at almost equal levels, and the adhesion was enhanced 20 to 50% when the cells were precultured with TNF-alpha for 24 hr. The inhibition assay using either (anti-ICAM-1 + anti-LFA-1, lymphocyte function-associated antigen-1) or (anti-VCAM-1 + anti-VLA-4, very late antigen-4) mAbs demonstrated that the ICAM-1 system was utilized more preferentially by T than B blasts when F-2 cells were stimulated with TNF-alpha, and the VCAM-1 system was vice versa under the unstimulated and stimulated conditions. Granulocytes also adhered to F-2 cells, but no mAbs could inhibit the adhesion. Although F-2 cells produced a considerable amount of IL-6, GM-CSF and neutrophil chemotactic activity, a 24 hr incubation with TNF-alpha resulted in an increase of 12 fold in IL-6 and 3 fold in neutrophil chemotactic activity production.
