[Effect of transfected Rv0901 gene on the activity of mice macrophages].
OBJECTIVE
To test the effect of Rv0901 gene of Mycobacterium tuberculosis on the activity of mice macrophages.


METHODS
Peritoneal macrophages of mice were isolated and transfected with pcDNA3. 1 or pcDNA3. 1-Rv0901 plasmid DNA, along with the GFP DNA. The effectiveness of the transfection was detected by RT-PCR. The expression of the GFP and the apoptosis ratio of the macrophages were detected by FCM 72 hours after the transfection. The level of nitric oxide and IFN-gamma in cultural supernatant were also measured 72 hours after transfection.


RESULTS
All of the macrophages being transfected had the expression of GFP. After being transfected with pcDNA3. 1-Rv0901, the 528 bp gene of Rv0901 was amplified by RT-PCR. The macrophages transfected with pcDNA3. 1-Rv0901 had higher apoptosis ratio [(56.4 +/- 2.0)% vs (19.9 +/- 1.5)%] and released more nitric oxide [(40.4 +/- 3.0) micromol/L vs (27.5 +/- 3.2) micromol/L] and IFN-gamma [(2.11 +/- 0.031) ng/mL vs (0.62 +/- 0.025) ng/mL] in the cultural supernatants than those transfected with pcDNA3. 1 (P < 0.05).


CONCLUSIONS
Transient transfection of pcDNA3. 1-Rv0901 increases the apoptosis ratio of the macrophages, which could increase the release of nitric oxide and IFN-gamma.
Zhong
Qi
Q
Department of Infection and Immunity, West China school of Preclinical and Forensic Medicine, Sichuan University, Chengdu 610041, China.
Bao
Lang
L
Shang
Zheng-Ling
ZL
Liu
Yu
Y
Huang
Bi
B
Yao
Su-Xia
SX
D000818:Animals; D017209:Apoptosis / Q000235:genetics; D001426:Bacterial Proteins / Q000235:genetics*; D005260:Female; D005822:Genetic Vectors / Q000235:genetics; D007371:Interferon-gamma / Q000378:metabolism; D008264:Macrophages / Q000166:cytology / Q000378:metabolism*; D051379:Mice; D009569:Nitric Oxide / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013997:Time Factors; D014162:Transfection; D037521:Virulence Factors / Q000235:genetics*
D001426:Bacterial Proteins; C539381:Rv0901 protein, Mycobacterium tuberculosis; D037521:Virulence Factors; D009569:Nitric Oxide; D007371:Interferon-gamma