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Updated: Sep 28, 2026
Generation of Human CD40-activated B cells
Published on: October 16, 2009
[CD40 ligandization promotes IL-6 and Flt3 ligand production of bone marrow stromal cells]
Jinxiang Fu1, Yan Liu, Xueguang Zhang
1Hematology Department, Second Affiliated Hospital of Soochow University, Suzhou 215004, China.
Insights
Stimulating CD40 on bone marrow stromal cells (BMSC) increases IL-6 and FL production, promoting hematopoietic stem cell proliferation and survival.
Area of Science:
- Hematology
- Immunology
- Cell Biology
Background:
- Bone marrow stromal cells (BMSC) play a crucial role in hematopoiesis.
- The CD40/CD40L pathway is implicated in immune responses and cell-cell interactions.
Purpose of the Study:
- To investigate CD40 and CD40L expression on BMSC.
- To analyze the impact of CD40 stimulation on cytokine production by BMSC.
- To determine the role of stimulated BMSC in regulating hematopoiesis.
Main Methods:
- BMSC were isolated and analyzed for CD40 expression via flow cytometry.
- Cytokine levels (IL-3, IL-6, FL, SCF) in BMSC supernatant were measured using ELISA after stimulation with anti-CD40 antibody (5C11).
- The effect of stimulated BMSC supernatant on CD34+ cell proliferation and apoptosis was assessed using colony formation and Annexin V assays.
Main Results:
- BMSC express CD40.
- Stimulation with 5C11 increased IL-6 and FL production, with no significant change in IL-3 or SCF.
- Supernatant from stimulated BMSC enhanced CD34+ cell proliferation and CFU-GM formation, while reducing apoptosis.
Conclusions:
- CD40 ligandization on BMSC upregulates IL-6 and FL, promoting CD34+ cell proliferation.
- The CD40/CD40L interaction on BMSC may regulate hematopoiesis through cytokine network modulation in the bone marrow.
Objective:
To analyze the expression of the CD(40) and its ligand (CD(40)L) on bone marrow stromal cells (BMSC), and investigate interleukin-3 (IL-3), IL-6, Flt3 ligand (FL) and stem cell factor (SCF) production of BMSC after stimulated with CD(40) agonistic monoclonal antibody (5C11) and its role in the regulation of hematopoiesis.
Methods:
BMSC were freshly isolated from adult bone marrow. The expression of CD(40) on these cells was determined with flow cytometry, the concentrations of IL-3, IL-6, FL and SCF in the supernatant at 24, 48 and 72 hours after BMSC cultured with 5C11 at a dose of 20 micro g/ml were determined by the ELISA assay. After BMSC incubated with 5C11 for 24 hours, the supernatant was collected and cultured with purified cord blood CD(34)(+) cells for 7 days under 37 degrees C in a fully humidified atmosphere supplement with 5% CO(2). Colony formation assay and Annexin V assay were employed to determine the proliferation and apoptosis of the CD(34)(+) cells.
Results:
BMSC expressed CD(40) and the production of IL-6 and FL increased after stimulated with 5C11 while IL-3 and SCF had no change. The supernatant collected from the stimulated BMSC promoted proliferation of CD(34)(+) cells, increased the CFU-GM yields and had anti-apoptosis effects.
Conclusion:
CD(40) ligandization on BMSC increased the production of IL-6 and FL and promoted the proliferation of CD(34)(+) cells. The couple CD(40)/CD(40)L may be involved in the control of hematopoiesis via modulation of the cytokine network in the bone marrow.
