Concurrent CCR7 Overexpression and RelB Knockdown in Immature Dendritic Cells Induces Immune Tolerance and Improves Skin-Graft Survival in a Murine Model

Cell Physiol Biochem. 2017;42(2):455-468. doi: 10.1159/000477593. Epub 2017 Jun 5.

Abstract

Background/aims: Skin transplantation aims to cover skin defects but often fails due to immune rejection of the transplantated tissue. Immature dendritic cells (imDCs) induce immune tolerance but have a low migration rate. After stimulation, imDCs transform into mature DCs, which activate immune rejection. Thus, inducing imDC to obtain a high migration counteracts development of immune tolerance.

Methods & results: We transfected imDCs with a recombinant adenovirus carrying the CCR7 gene (Ad-CCR7) and a small interfering RNA targeting RelB (RelB-siRNA) to concurrently overexpress CCR7 and downregulate RelB expression. Functionally, such cells showed a significantly enhanced migration rate in the chemotactic assay and decreased T-cell proliferation after lipopolysaccharide stimulation in mixed lymphocyte reactions. Cotransfected cells showed an increased ability to induce immune tolerance by upregulating T regulatory (Treg) cells and shifting the Th1/Th2 ratio. Cotransfection of Ad-CCR7 and RelB-siRNA endowed imDCs with resistance to apoptosis and cell death. CCR7 overexpression and RelB knockdown (KD) in imDCs improve skin-graft survival in a murine skin-transplantation model.

Conclusion: Transfection with Ad-CCR7 and RelB KD in imDCs may be an effective approach inducing immune tolerance, thus being potentially valuable for inhibiting allograft rejection.

Keywords: CCR7; Immune tolerance; RelB; Transplantation; Treg cells; imDCs.

MeSH terms

  • Adenoviridae
  • Animals
  • Dendritic Cells / immunology
  • Dendritic Cells / metabolism
  • Gene Expression Regulation
  • Graft Rejection / genetics
  • Graft Rejection / immunology
  • Graft Survival / genetics
  • Humans
  • Immune Tolerance / genetics*
  • Mice
  • Receptors, CCR7 / biosynthesis*
  • Receptors, CCR7 / genetics
  • Skin / immunology*
  • Skin / metabolism
  • T-Lymphocytes, Regulatory / immunology
  • T-Lymphocytes, Regulatory / metabolism
  • Transcription Factor RelB / genetics*
  • Transfection

Substances

  • Ccr7 protein, mouse
  • Receptors, CCR7
  • Relb protein, mouse
  • Transcription Factor RelB