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Published on: April 18, 2016
[Effect of soluble anti-CD47 monoclonal antibody on the differentiation and function of human dendritic cells]
Mao-fang Lin1, Jing Yu, Li-xing Yan
1The First Affiliated Hospital, Medical College of Zhejiang University, Hangzhou 310003, China.
Insights
The anti-CD47 monoclonal antibody (mAb) B6H12 inhibits the maturation and function of dendritic cells (DCs). This study reveals a negative impact of CD47 blockade on DC immune responses.
Area of Science:
- Immunology
- Cell Biology
Background:
- Dendritic cells (DCs) are crucial antigen-presenting cells in the immune system.
- CD47 is a molecule expressed on DCs that plays a role in immune regulation.
Purpose of the Study:
- To investigate the effect of anti-CD47 monoclonal antibody (mAb) B6H12 on the differentiation and function of cultured human dendritic cells (DCs).
Main Methods:
- Human peripheral monocyte-derived DCs were cultured with GM-CSF, LPS, and IL-4, with or without soluble B6H12.
- Immunophenotypes were analyzed by flow cytometry.
- mRNA and protein expression of IL-12 were assessed using RT-PCR and ELISA.
- Antigen-presenting function was evaluated via a mixed leukocyte reaction assay.
Main Results:
- High CD47 expression (94-98%) was observed on DCs.
- B6H12 treatment significantly reduced the expression of DC maturation markers (CD80, CD86, CD83, CD1a, HLA-DR).
- IL-12 production and DC antigen-presenting capacity were significantly inhibited by B6H12 mAb.
Conclusions:
- Anti-CD47 mAb B6H12 negatively impacts the maturation and function of cultured dendritic cells.
- CD47 blockade may impair DC-mediated immune responses.
Objective:
To explore the influences of anti-CD47 monoclonal antibody (mAb) B6H12 on the differentiation and function of cultured dendritic cells (DCs).
Methods:
Human peripheral monocyte derived DCs were propagated with granulocyte-macrophage colony-stimulating factor (GM-CSF) and lipopolysaccharide (LPS) plus interleukin-4 (IL-4) in the presence or absence of soluble B6H12. Flow cytometry was used to analyze the immunophenotypes of cells, semi-quantitative RT-PCR and ELISA methods to analyse the mRNA and protein expression levels of interleukin-12 (IL-12). The antigen-presenting function of DCs was determined in one-way mixed leukocyte reaction by Brdu-ELISA technique.
Results:
There was a high expression rate (94% approximately 98%) of CD47 molecules in DCs. The cell immunophenotypes in B6H12 mAb treated and untreated DC groups were as follows: CD80(+) (68.14 +/- 7.41)% vs (89.17 +/- 8.59)%; CD86(+) (67.33 +/- 4.71)% vs (87.27 +/- 3.56)%; CD83 (40.08 +/- 14.80)% vs (72.77 +/- 8.68)%; CD1a(+) (66.45 +/- 4.06)% vs (95.93 +/- 3.03)%; and HLA-DR (40.67 +/- 13.48) vs (98.97 +/- 1.01)%, respectively. The expression levels of mRNA and protein of IL-12 were strongly inhibited in B6H12 mAb treated DC (P < 0.01). The quantity of Brdu was also lower in B6H12 mAb treated DC than that in untreated DCs (P < 0.01).
Conclusion:
The anti-CD47 McAb exerts a negative effect on the maturation and functions of cultured DCs.
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