[Retroviral vector-mediated in vitro expression of human soluble fas]

Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2002 Apr;10(2):97-9.
[Article in Chinese]

Abstract

Leukemic cells from patients expressed high level FasL cause apoptosis of autologous activated T cells via the Fas/FasL pathway. To investigate the role of soluble Fas (sFas) in reversing this process, a retroviral-mediated expression vector pLXIN-sFas was established. A retroviral-mediated expression system of human sFas was established in vitro and the biological activity of the expression product sFas was observed. To obtain the soluble Fas cDNA, the specific part of the full-length Fas cDNA was deleted by multiple PCR. After pLXIN-sFas packaged by PA317 cells, it was transferred into the target cell COS-7. The quantity of the sFas was (2.2 +/- 0.7) micro g/ml in supernatant of cultured COS-7 cells, and it could greatly inhibit apoptosis of Jurket cells induced by anti-Fas antibody. Our results suggested that the recombinant is able to express the target proteins in vitro and it has the perfect biological activity.

MeSH terms

  • 3T3 Cells
  • Animals
  • Apoptosis / drug effects
  • COS Cells
  • Cell Line
  • Cell Survival / drug effects
  • Culture Media, Conditioned / pharmacology
  • Dose-Response Relationship, Drug
  • Gene Expression
  • Genetic Vectors / genetics
  • Humans
  • Jurkat Cells
  • Mice
  • Retroviridae / genetics*
  • Solubility
  • fas Receptor / genetics*
  • fas Receptor / metabolism
  • fas Receptor / pharmacology

Substances

  • Culture Media, Conditioned
  • fas Receptor