[Construction of recombinant adenovirus expressing sCD40L-Ig]

Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2005 Nov;21(6):668-71.
[Article in Chinese]

Abstract

Aim: To construct adenovirus expressing secretory human CD40L-Ig (sCD40 ligand Ig fusion protein).

Methods: The genes encoding the extracellular domain of human CD40L and IgG Fc were amplified with PCR and inserted into shuttle vector pAdTrack-CMV which then was transformed into E. coli pAdEasy-1-BJ5183 to produce recombinant Ad plasmid. The recombinant plasmid was digested with Pac I and transfected into 293 cells to generate recombinant adenovirus. The recombinant adenovirus was then used in mixed lymphocyte reaction (MLR) to test its function.

Results: The recombinant adenovirus sCD40L-Ig was produced. It could inhibit the lymphocyte proliferation in MLR.

Conclusion: The sCD40L-Ig adenovirus is prepared successfully and its inhibition of MLR is confirmed.

Publication types

  • English Abstract

MeSH terms

  • Adenoviridae / genetics*
  • Animals
  • Blotting, Western
  • Cell Line
  • Cell Proliferation / drug effects
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Humans
  • Lymphocyte Culture Test, Mixed
  • Lymphocytes / cytology
  • Lymphocytes / drug effects
  • Mice
  • Mice, Inbred BALB C
  • Mice, Inbred C57BL
  • Polymerase Chain Reaction
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism*
  • Recombinant Fusion Proteins / pharmacology*

Substances

  • CD40LIg fusion protein
  • Recombinant Fusion Proteins
  • Green Fluorescent Proteins