Immunoregulation effects in vitro of the xenoprotein in combination with recombinant human granulocyte-macrophage colony stimulating factor and bacillus Calmette-Guerin.
- Author:
Ming-Li WANG
1
;
Zhi-Gang XIE
;
Han LU
;
Ming SHI
;
Mei-Ru HU
;
Ming YU
;
Yuan-Fang MA
;
Bei-Fen SHEN
;
Ning GUO
Author Information
1. Laboratory of Cytoimmunology, Institute of Basic Medical Sciences, Academy of Military Medical Sciences, Beijng 100850, China.
- Publication Type:Journal Article
- MeSH:
Animals;
BCG Vaccine;
immunology;
CHO Cells;
Cricetinae;
Cricetulus;
Granulocyte-Macrophage Colony-Stimulating Factor;
immunology;
Humans;
Interleukin-10;
metabolism;
Interleukin-12;
metabolism;
Rats;
Recombinant Proteins;
immunology;
Th1 Cells;
immunology;
Th2 Cells;
immunology
- From:
Journal of Experimental Hematology
2008;16(6):1408-1412
- CountryChina
- Language:Chinese
-
Abstract:
This study was aimed to investigate the effects of xenogeneic antigen neu-Fc in combination with the recombinant human granulocyte-macrophage colony stimulating factor (GM-CSF) and Bacillus Calmette-Guerin (BCG) on the regulation of Th1 and Th2 immune response in vitro. The rat neu L2-S2 domain was engineered as a chimeric protein with human IgG Fc. The eukaryotic expression vector was constructed. The recombinant protein was stably expressed in CHO cells and purified by rProtein A Sepharose Fast Flow column. The recombinant protein was identified by SDS-PAGE and Western blot. Peripheral blood mononuclear cells (PBMNCs) were obtained by means of standard Ficoll separation from the blood of healthy donors. Neu-Fc-induced PBMNC proliferation was tested by MTT. The production of IL-12 and IL-10 was measured by ELISA. The results showed that the level of IL-12 decreased and IL-10 increased after PBMNCs were incubated with MCF-7 cultural supernatant. 10 nmol/L neu-Fc strongly induced the cell proliferation. Compared with neu-Fc or GM-CSF or BCG treatment alone, neu-Fc in combination with GM-CSF and BCG significantly stimulated IL-12 production and inhibited IL-10 production (p < 0.01). It is concluded that the neu-Fc can stimulate the proliferation activity of PBMNCs. neu-Fc, GM-CSF and BCG costimulation efficiently induces Th1 immune response.
