Role of IFN-γ in the regulation of murine hemopoiesis / 中国实验血液学杂志
Journal of Experimental Hematology
;
(6): 612-616, 2014.
Article
in Chinese
| WPRIM
| ID: wpr-349661
ABSTRACT
This study was aimed to explore the role and mechanism of IFN-γ in the regulation of hemopoiesis in mice. Murine IFN-γ fragment was amplified from murine splenic cells with RT-PCR and plasmid pCDH1-mIFN-γ-EF1-copGFP (pCDH-mIFN-γ-GFP) was constructed. Plasmids pCDH-mIFN-γ-GFP and pCDH1-EF1-copGFP (pCDH-GFP) together with packaging plasmids pPACK-A, pPACK-B and pPACK-C were respectively transfected into 293T cells by using a method of calcium phosphate precipitation to produce lentivirus. Bone marrow mononuclear cells (BMMNC) from male C57BL/6J mice were transfected with the lentiviral vector pCDH expressing mIFN-γ and green fluorescent protein (GFP). The cells were cultured in M3434 semi-solid medium for colony formation assay and transplanted into lethally-irradiated mice through caudal vein injection, and the peripheral blood cell counts and GFP were monitored regularly after transplantation. The results showed that lentiviral vector pCDH-mIFN-γ-GFP was constructed successfully and 293T cells transfected with mIFN-γ secreted mIFN-γ. Transfection of mIFN-γ into BMMNC decreased colony formation, colony number of the mIFN-γ group was significantly less than that of the control group. The recovering of circulating blood cell parameters in mIFN-γ transplantation group was significantly later than control group. GFP positive cells could be detected in the peripheral blood at 8 weeks after transplantation. It is concluded that mIFN-γ may inhibit the colony-forming capacity of transduced BMMNC and delay the hematopoietic reconstitution.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Pharmacology
/
Plasmids
/
Bone Marrow Cells
/
Transfection
/
Cell Line
/
Interferon-gamma
/
Lentivirus
/
Cell Biology
/
Genetic Vectors
/
Genetics
Limits:
Animals
Language:
Chinese
Journal:
Journal of Experimental Hematology
Year:
2014
Type:
Article
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