Donor MHC gene to mitigate rejection of transplantation in recipient mice

Chin Med J (Engl). 2011 Dec;124(24):4279-85.

Abstract

Background: Donor organ rejection continues to be a significant problem for patients receiving transplants. We therefore tested whether transferring a donor's major histocompatibility complex (MHC) gene to the recipient would mitigate the rejection of transplanted hearts in mice.

Methods: H-2K(k) gene from donor mice was amplified using nested polymerase chain reaction (PCR) and ligated into a mammalian expression vector, which was then transfected into thymus ground mass cells collected from the recipients. Clones stably expressing the transgene were then injected into the recipients' thymus visualized using ultrasound. Control mice were administered cells previously transfected with empty vector. Following heart transplantation, cardiac activity was monitored electrocardiographically. Recipient thymus cells were tested for MHC antigenicity using flow cytometry and spleen cells were subjected to mixed lymphocyte culture tests. Finally, the transplanted hearts were sectioned, stained and examined under light microscopy.

Results: Southern analysis following nested PCR revealed clear expression of H-2K(k) gene. Following transplantation, electrocardiosignals were detectable highly significantly longer in recipients administered thymal cells expressing donor H-2K(k) than in those receiving control cells. Flow cytometric analysis using an anti-H-2K(k) antibody confirmed its expression in H-2K(k) treated recipients but not in control mice. Mixed lymphocyte cultures containing H-2K(k) treated cells showed significantly less proliferation than those containing control cells. Hearts from control mice showed substantially greater lymphocyte infiltration than those from H-2K(k) treated mice and large areas of necrosis.

Conclusion: Rejection of transplanted hearts can be mitigated substantially by introducing the donor's MHC into the recipient.

MeSH terms

  • Animals
  • Blotting, Southern
  • Electrocardiography
  • Female
  • Flow Cytometry
  • Graft Rejection / genetics
  • Graft Rejection / immunology
  • Heart Transplantation / immunology*
  • Heart Transplantation / methods*
  • Major Histocompatibility Complex / genetics*
  • Major Histocompatibility Complex / immunology
  • Male
  • Mice
  • Polymerase Chain Reaction