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Updated: Jun 2, 2026
Flow Cytometry Analysis of Tissue Factor Expression in Human Platelets
Published on: November 22, 2024
[Activation of tumor necrosis factor receptor-associated factor 6 in anti-β2GPI/β2GPI-induced tissue factor
Guo-ying Xu1, Hong Zhou, Hai-ping Wen
1School of Medical Science and Laboratory Medicine, Jiangsu University, Zhenjiang 212013, China. peakxuguoying@126.com
Aim:
To investigate whether tumor necrosis factor receptor-associated factor 6 (TRAF6) is involved in anti-β2GPI/β2GPI-induced tissue factor (TF) expression on THP-1 cells.
Methods:
The total RNA was extracted and the protein lysates were collected from THP-1 cells stimulated with anti-β2GPI/β2GPI complex. And then the TF expression on THP-1 cells was detected by real-time quatitative PCR (RT-qPCR) and TF activity kit. TRAF6 mRNA and its protein expression were investigated by RT-qPCR and Western blotting, respectively. The proteasome inhibitor, MG-132, was used for inhibitory assays, in order to demonstrate the effect of anti-β2GPI/β2GPI complex on THP-1 cells.
Results:
The TF expression (both mRNA and activity) on THP-1 cells was significantly up-regulated with the treatment of anti-β2GPI/β2GPI complex (100 mg/L), compared with untreated cells(P<0.05). The TRAF6 mRNA and protein levels in THP-1 cells were also significantly increased with the treatment of anti-β2GPI/β2GPI complex. The expression of TRAF6 was shown in a time-dependent manner, with the maximal level at 15 minutes (mRNA) and 30 minutes (protein) respectively. All the stimulating effects of anti-β2GPI/β2GPI complex (100 mg/L) on THP-1 cells were inhibited by MG-132 (5 μmol/L).
Conclusion:
TRAF6 is up-regulated and contributes to TF expression on THP-1 cells induced with anti-β2GPI/β2GPI complex.
Insights
Tumor necrosis factor receptor-associated factor 6 (TRAF6) is upregulated by anti-β2GPI/β2GPI complex in THP-1 cells. This TRAF6 upregulation contributes to the increased tissue factor (TF) expression.
Area of Science:
- Immunology
- Molecular Biology
- Cell Biology
Context:
- Antiphospholipid syndrome is associated with thrombosis.
- Beta2-glycoprotein I (β2GPI) is a key autoantigen in antiphospholipid syndrome.
- Tissue factor (TF) plays a crucial role in the coagulation cascade and thrombosis.
Purpose:
- To investigate the role of tumor necrosis factor receptor-associated factor 6 (TRAF6) in anti-β2GPI/β2GPI-induced tissue factor (TF) expression.
- To determine if TRAF6 is upregulated by anti-β2GPI/β2GPI complex in THP-1 cells.
- To elucidate the mechanism by which anti-β2GPI/β2GPI complex induces TF expression.
Summary:
- Anti-β2GPI/β2GPI complex significantly up-regulated TF expression (mRNA and activity) in THP-1 cells.
- TRAF6 mRNA and protein levels were also significantly increased in a time-dependent manner.
- The proteasome inhibitor MG-132 blocked the effects of anti-β2GPI/β2GPI complex, indicating TRAF6 involvement.
Impact:
- This study identifies TRAF6 as a key mediator in the prothrombotic effects of anti-β2GPI antibodies.
- Understanding the TRAF6 pathway may lead to novel therapeutic strategies for antiphospholipid syndrome.
- Provides insights into the molecular mechanisms linking autoimmunity to thrombosis.
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