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Published on: October 16, 2009
CD40 ligand increases expression of its receptor CD40 in human coronary artery endothelial cells
Hong Chai1, Shaoyu Yan, Hao Wang
1Molecular Surgeon Research Center, Division of Vascular Surgery and Endovascular Therapy, Michael E. DeBakey Department of Surgery, Baylor College of Medicine, Houston, TX 77030, USA.
Insights
CD40 ligand (CD40L) increases its receptor CD40 expression in human coronary artery endothelial cells. This CD40L-induced upregulation involves oxidative stress and ERK1/2 activation, potentially contributing to vascular disease.
Area of Science:
- Cardiovascular Biology
- Endothelial Cell Function
- Molecular Mechanisms of Atherosclerosis
Background:
- CD40 ligand (CD40L) and its receptor CD40 are implicated in atherosclerosis.
- Elevated CD40L levels correlate with increased cardiovascular event risk.
Purpose of the Study:
- To investigate if CD40L upregulates its receptor CD40 expression in human coronary artery endothelial cells (HCAECs).
- To explore potential feedback mechanisms by which CD40L enhances its function.
Main Methods:
- HCAECs were treated with soluble CD40L.
- CD40 mRNA and protein levels were quantified using real-time PCR and Western blot.
- Involvement of oxidative stress and MAPK pathways (ERK1/2) was assessed using inhibitors and antioxidants.
Main Results:
- CD40L treatment significantly increased CD40 mRNA (79%) and protein (80%) levels in HCAECs.
- The increase was specifically blocked by anti-CD40L antibody.
- Oxidative stress (SeMet) and ERK1/2 inhibition (PD98059) blocked CD40L-induced CD40 upregulation.
Conclusions:
- Clinically relevant CD40L concentrations upregulate CD40 expression in HCAECs.
- This upregulation appears mediated by oxidative stress and ERK1/2 activation.
- Suggests a novel mechanism for CD40L enhancing vascular function and contributing to endothelial dysfunction.
Background:
Recently, CD40 ligand (CD40L) and its receptor CD40 have been implicated in atherosclerosis. Clinical data showed that elevated CD40L levels are associated with a high risk of cardiovascular events. The aim of this study was to investigate whether CD40L could affect the expression of its membrane receptor CD40 as a feedback mechanism by which CD40L could enhance its functions in human coronary artery endothelial cells (HCAECs).
Methods:
The HCAECs were treated with human soluble CD40L, and the messenger RNA (mRNA) and protein levels of CD40 were determined by real-time polymerase chain reaction and Western blot analysis, respectively. The specific effect of CD40L was confirmed by a blocking experiment with antibody against CD40L. Involvements of oxidative stress and mitogen-activated protein kinases (MAPKs) were also studied with antioxidant seleno-L-methionine (SeMet) and MAPK inhibitors such as extracellular signal-regulated kinase 1/2 (ERK1/2) inhibitor.
Results:
When HCAECs were cultured with CD40L (5 microg/mL) for 24 hours, CD40 mRNA levels were increased by 79% compared with controls (P < .05). Similarly, Western blot analysis showed an 80% increase in CD40 protein levels (P < .05). The CD40L-induced increase in CD40 mRNA levels were blocked specifically by anti-CD40L antibody. Antioxidant SeMet and specific ERK1/2 inhibitor (PD98059) also effectively blocked CD40L-induced CD40 mRNA increase.
Conclusions:
These data demonstrate that clinically relevant concentration of CD40L increased the expression of its receptor CD40 in HCAECs. The CD40L-induced upregulation of CD40 may be mediated by oxidative stress and ERK1/2 activation. This study suggests a new mechanism by which CD40L could enhance its biologic functions in the vascular system and contribute to endothelial dysfunction and vascular disease.
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