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
em Zh
| WPRIM
| ID: wpr-349661
Biblioteca responsável:
WPRO
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.
Texto completo:
1
Índice:
WPRIM
Assunto principal:
Farmacologia
/
Plasmídeos
/
Células da Medula Óssea
/
Transfecção
/
Linhagem Celular
/
Interferon gama
/
Lentivirus
/
Biologia Celular
/
Vetores Genéticos
/
Genética
Limite:
Animals
Idioma:
Zh
Revista:
Journal of Experimental Hematology
Ano de publicação:
2014
Tipo de documento:
Article