Lentiviral-mediated shRNA against RelB induces the generation of tolerogenic dendritic cells

Int Immunopharmacol. 2012 Mar;12(3):501-9. doi: 10.1016/j.intimp.2012.01.004. Epub 2012 Jan 20.


Objective: Lentiviral-mediated shRNA against RelB was used to produce tolerogenic dendritic cells from murine bone marrow derived dendritic cells (BMDCs).

Method: RelB expression in the BMDCs was silenced by lentivirus carrying RelB shRNA. The apoptosis rate and surface markers of DCs were assessed by flow cytometry. IL-12,IL-10,TGF-β1 secreted by DCs and DNA binding capacity of NF-κB subunits in the nucleus were measured by ELISA, independently. MLR was used to analyze the capacity of DCs to inhibit immune response.

Results: RelB expression was significantly inhibited in DCs following lentiviral mediated delivery of RelB specific shRNA. The RelB shRNA-DC produced lower IL-12 and higher IL-10 than mature dendritic cells (mDCs) and silencing control DCs. There was no difference in the apoptosis rate between shRNA RelB-DCs and mDCs. The expression levels of co-stimulatory molecules (CD80, CD86 and CD83) and MHC-II class molecule were lower in the RelB shRNA-DCs than in the mDCs and silencing control DCs. In addition, RelB shRNA also inhibited the RelB DNA binding capacity but had no effect on other NF-κB subunits. The shRNA RelB-DCs can significantly inhibit mixed lymphocyte reaction (MLR) and down-regulate Th1 cytokines and prompt the production of Th2 cytokines.

Conclusion: Our results indicate RelB shRNA transfection of DCs can induce the immature status, and produce tolerogenic DCs.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Apoptosis / genetics
  • Blotting, Western
  • Bone Marrow Cells / metabolism
  • DNA-Binding Proteins / metabolism
  • Dendritic Cells / drug effects*
  • Flow Cytometry
  • Gene Silencing
  • Genetic Vectors
  • Interleukin-10 / biosynthesis
  • Interleukin-10 / genetics
  • Interleukin-12 / biosynthesis
  • Interleukin-12 / genetics
  • Lentivirus / genetics*
  • Lymphocyte Culture Test, Mixed
  • Membrane Proteins / analysis
  • Membrane Proteins / metabolism
  • Mice
  • Mice, Inbred C57BL
  • NF-kappa B / biosynthesis
  • NF-kappa B / genetics
  • RNA, Small Interfering / genetics*
  • Real-Time Polymerase Chain Reaction
  • Th1 Cells / drug effects
  • Th1 Cells / metabolism
  • Th2 Cells / drug effects
  • Th2 Cells / metabolism
  • Transcription Factor RelB / biosynthesis*
  • Transcription Factor RelB / genetics*


  • DNA-Binding Proteins
  • Membrane Proteins
  • NF-kappa B
  • RNA, Small Interfering
  • Interleukin-10
  • Transcription Factor RelB
  • Interleukin-12