Chinese Journal of Tissue Engineering Research ›› 2012, Vol. 16 ›› Issue (18): 3337-3340.doi: 10.3969/j.issn.1673-8225.2012.18.024

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Transfection of rat CD4+CD25- T cells with lentiviral vector containing Foxp3 gene and its expression

Huang Yang1, Li Yong-xiang1, Shen Ji-long2, Luo Qing-li2, Lu Wei2, Huo Xing-xing2   

  1. 1First Affiliated Hospital of Anhui Medical University, Hefei  230032, Anhui Province, China; 2 Key Laboratory of Microbiology and Parasitology (Anhui Medical University), the Key Laboratory of Zoonoses (Anhui Medical University), Hefei  230032, Anhui Province, China
  • Received:2011-12-31 Revised:2012-01-25 Online:2012-04-29 Published:2012-04-29
  • Contact: Li Yong-xiang, Doctor, Professor, Chief physician, Doctoral supervisor, First Affiliated Hospital of Anhui Medical University, Hefei 230032, Anhui Province, China yongxiangli@yahoo.com.cn
  • About author:Huang Yang★, Studying for master’s degree, First Affiliated Hospital of Anhui Medical University, Hefei 230032, Anhui Province, China yanghuang6699@yahoo.com.cn
  • Supported by:

    the National Natural Science Foundation of China, No.30871207*; Key Scientific Research Projects of Anhui Science and Technology Agency, No.7020304101*; Key Scientific Research Projects of Anhui Education Department, No.KJ2008A154*

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Abstract:

BACKGROUND: Regulatory T cells play an important role in maintaining immune response homeostasis and immune tolerance, but the CD4+CD25+T cells are less in peripheral blood and with low proliferation ability in vitro.
OBJECTIVE: To observe the expression of transfected rat Foxp3 gene in rat CD4+CD25- T cells using lentiviral vector carrier EGFP+.
METHODS: Rat CD4+CD25-T cells were sorted by magnetic activated cell sorting, and transfected by lentiviral vector containing rat Foxp3 gene in vitro. The whole observation was divided into three groups. Transfected rat CD4+CD25- T cells as the experimental group, the blank plasmid EGFP+ and rat CD4+CD25-T cells as the negative control group, rat CD4+CD25+T cells as the positive control group. The expression of Foxp3 gene was assayed by fluorescence microscope and RT-PCR from the level of protein and mRNA respectively.
RESULTS AND CONCLUSION: The high purity CD4+CD25- and CD4+CD25+ T cells were obtained and the survival rate of cells was (94±2)%. Significantly high expression of rat Foxp3 gene was detected in lentiviral vector transfected CD4+CD25- T cells. The lentiviral vector system carrying rat Foxp3 gene can efficiently mediate the highly expression of rat Foxp3 gene in the rat CD4+CD25- T cells.

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