Role of IFN-γ in the regulation of murine hemopoiesis.
10.7534/j.issn.1009-2137.2014.03.007
- Author:
Xin ZHAO
1
;
Hai-Yan XING
1
;
Zheng TIAN
1
;
Ke-Jing TANG
1
;
Min WANG
1
;
Qing RAO
1
;
Jian-Xiang WANG
2
;
Feng-Kui ZHANG
3
Author Information
1. State Key Laboratory of Experimental Hematology, Institute of Hematology & Blood Disease Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Tianjin 300020, Tianjin Province, China.
2. State Key Laboratory of Experimental Hematology, Institute of Hematology & Blood Disease Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Tianjin 300020, Tianjin Province, China. E-mail: wangjx@ihcams.ac.cn.
3. State Key Laboratory of Experimental Hematology, Institute of Hematology & Blood Disease Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Tianjin 300020, Tianjin Province, China. E-mail: zhfk@hotmail.com.
- Publication Type:Journal Article
- MeSH:
Animals;
Bone Marrow Cells;
cytology;
drug effects;
Cell Line;
Genetic Vectors;
Hematopoiesis;
drug effects;
genetics;
Interferon-gamma;
genetics;
pharmacology;
Lentivirus;
genetics;
Male;
Mice;
Mice, Inbred C57BL;
Plasmids;
Transfection
- From:
Journal of Experimental Hematology
2014;22(3):612-616
- CountryChina
- Language:Chinese
-
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.
