A Mouse Model of CMV Transmission Following Kidney Transplantation

Abstract

Reactivation of latent CMV in transplant recipients remains a significant infectious complication of transplantation. Investigation of the cellular and molecular mechanisms by which reactivation occurs has been hampered by the lack of appropriate animal models. Here, we show that transplantation of kidneys latently infected with murine cytomegalovirus (MCMV) into NOD.Cg-Prkdc scid IL2rg tm1Wjl /Szj mice results in reactivation of latent virus in the kidney, resulting in a disseminated primary infection of the recipient. This model will be useful in elucidating mechanisms of MCMV reactivation, including the roles of injury and of spontaneous reactivation, and in testing new therapies for treatment and prevention of CMV reactivation and disease.

Epigenetic Control of Cytomegalovirus Latency and Reactivation

“…A new model for reactivation of infectious virus in genetically immunodeficient transplant recipients has recently been described [85]. As in the previous model, activation of transcription factors that bind to the ie promoter and reactivation of ie gene expression was detectable within two days after transplant (unpublished observations), but unlike the allogeneic transplant model, reactivation of infectious virus becomes detectable in immunodeficient recipients over a period of weeks.…”

“…This was likely due to the very slow rate of CMV replication combined with the rapid rejection that occurs when allogeneic organs are transplanted into immunocompetent mice. Subsequent studies showed that reactivation of infectious virus in the donor kidney, and spread of the virus to recipient organs, could be induced by transplanting latently infected kidneys into immunodeficient recipient mice [85]. …”

“…Thus, transcriptional reactivation of ie gene expression would be expected to lead to activation of transcription and remodeling of other regions of viral chromatin. Due to the difficulty in obtaining sufficient amounts of material for analysis of MCMV DNA, which is present at a copy number of ~1 MCMV genome per 10,000 cellular genomes in latent kidneys [85], only the ie region has been analyzed in the allogeneic kidney transplant reactivation model at Day 2. However, previous studies showed that induction of ie gene expression was transient in this model, with a peak of expression at Day 2, and that genes representative of later stages of replication were not detectable [14].…”

Transplant-induced reactivation of murine cytomegalovirus immediate early gene expression is associated with recruitment of NF-κB and AP-1 to the major immediate early promoter

“…Fresh tissue was finely minced on ice, and the chromatin was fixed in 1 % formaldehyde as previously described (Liu et al, 2008(Liu et al, , 2010, and frozen in liquid nitrogen. Due to the very low MCMV DNA copy number in kidneys of latent mice [*1 copy of MCMV DNA per 10 000 cellular genomes (Li et al, 2012)] and the large amount of chromatin required for each ChIP, chromatin from 5-6 kidneys was pooled for each antibody. Frozen pellets of fixed tissue from 30-50 transplants were resuspended in hypotonic lysis buffer, and processed for immunoprecipitation as previously described (Liu et al, 2008(Liu et al, , 2010.…”

Systematic review of mouse kidney transplantation

“…Three studies have used the mouse kidney transplant model to investigate the effect of transplant‐associated viral infections . In mice infected with polyomavirus the allograft developed worse histological injury with increased numbers of CD8+ T cells compared with virus‐free recipients .…”