Effect of soluble anti-CD47 monoclonal antibody on the differentiation and function of human dendritic cells.
- Author:
Mao-fang LIN
1
;
Jing YU
;
Li-xing YAN
Author Information
- Publication Type:Journal Article
- MeSH: Adult; Antibodies, Monoclonal; pharmacology; Antigens, CD; analysis; Antigens, CD1; analysis; B7-2 Antigen; analysis; CD47 Antigen; analysis; immunology; Cell Differentiation; drug effects; Cell Proliferation; drug effects; Cells, Cultured; Dendritic Cells; cytology; drug effects; metabolism; Enzyme-Linked Immunosorbent Assay; Flow Cytometry; Granulocyte-Macrophage Colony-Stimulating Factor; pharmacology; Humans; Immunoglobulins; analysis; Interleukin-12; genetics; metabolism; Interleukin-4; pharmacology; Lipopolysaccharides; pharmacology; Membrane Glycoproteins; analysis; Monocytes; cytology; drug effects; metabolism; Reverse Transcriptase Polymerase Chain Reaction
- From: Chinese Journal of Hematology 2004;25(12):709-712
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo explore the influences of anti-CD47 monoclonal antibody (mAb) B6H12 on the differentiation and function of cultured dendritic cells (DCs).
METHODSHuman 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.
RESULTSThere 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).
CONCLUSIONThe anti-CD47 McAb exerts a negative effect on the maturation and functions of cultured DCs.
