Marker tolerant, immunocompetent animals as a new tool for regenerative medicine and long-term cell tracking.
BACKGROUND
Immune-mediated rejection of labeled cells is a general problem in transplantation studies using cells labeled with any immunogenic marker, and also in gene therapy protocols. The aim of this study was to establish a syngeneic model for long-term histological cell tracking in the absence of immune-mediated rejection of labeled cells in immunocompetent animals. We used inbred transgenic Fischer 344 rats expressing human placental alkaline phosphatase (hPLAP) under the control of the ubiquitous R26 promoter for this study. hPLAP is an excellent marker enzyme, providing superb histological detection quality in paraffin and plastic sections.


RESULTS
Transplantation of cells from hPLAP transgenic (hPLAP-tg) F344 rats into wild-type (WT) F344 recipients failed because of immune-mediated rejection. Here we show that this problem can be overcome by inducing tolerance to the marker gene by transplantation of bone marrow from hPLAP-tg F344 rats into WT F344 hosts after lethal irradiation, or by neonatal exposure of WT F344 rats to hPLAP-tg F344 cells. As proof-of-principle, we injected bone marrow cells (BMC) from hPLAP-tg rats into the knee joint of marker tolerant, bone marrow-transplanted WT rats, and found successful engraftment and differentiation of donor cells. In addition, hPLAP-tg BMC injected intravenously in neonatally tolerized WT F344 hosts could be traced in lymph nodes, 2 months post-injection.


CONCLUSIONS
In combination with the excellent marker hPLAP, marker tolerant animals may open up new perspectives for all experiments requiring long-term histological tracking of genetically labeled cells.
Odörfer
Kathrin I
KI
Institute of Pathophysiology, Dept. of Natural Sciences, University of Veterinary Medicine, Vienna, Austria. kathrin.odoerfer@vu-wien.ac.at <kathrin.odoerfer@vu-wien.ac.at>
Unger
Nina J
NJ
Weber
Karin
K
Sandgren
Eric P
EP
Erben
Reinhold G
RG
D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology*; D016026:Bone Marrow Transplantation / Q000379:methods*; D005819:Genetic Markers; D007121:Immunocompetence / Q000502:physiology*; D007963:Leukocytes, Mononuclear / Q000166:cytology* / Q000637:transplantation*; D023421:Models, Animal; D051381:Rats; D011916:Rats, Inbred F344; D044968:Regenerative Medicine / Q000379:methods
D005819:Genetic Markers