Induction of human blood group a antigen expression on mouse cells, using lentiviral gene transduction.
The ABO histo-blood group system is the most important antigen system in transplantation medicine, yet no small animal model of the ABO system exists. To determine the feasibility of developing a murine model, we previously subcloned the human alpha-1,2-fucosyltransferase (H-transferase, EC 2.4.1.69) cDNA and the human alpha-1,3-N-acetylgalactosaminyltransferase (A-transferase, EC 2.4.1.40) cDNA into lentiviral vectors to study their ability to induce human histo-blood group A antigen expression on mouse cells. Herein we investigated the optimal conditions for human A and H antigen expression in murine cells. We determined that transduction of a bicistronic lentiviral vector (LvEF1-AH-trs) resulted in the expression of A antigen in a mouse endothelial cell line. We also studied the in vivo utility of this vector to induce human A antigen expression in mouse liver. After intrahepatic injection of LvEF1-AH-trs, A antigen expression was observed on hepatocytes as detected by immunohistochemistry and real-time RT-PCR. In human group A erythrocyte-sensitized mice, A antigen expression in the liver was associated with tissue damage, and deposition of antibody and complement. These results suggest that this gene transfer strategy can be used to simulate the human ABO blood group system in a murine model. This model will facilitate progress in the development of interventions for ABO-incompatible transplantation and transfusion scenarios, which are difficult to develop in clinical or large animal settings.
Fan
Xiaohu
X
Department of Pediatrics, University of Alberta, Edmonton, AB, T6G 2E1 Canada.
Lang
Haili
H
Zhou
Xianpei
X
Zhang
Li
L
Yin
Rong
R
Maciejko
Jessica
J
Giannitsos
Vasiliki
V
Motyka
Bruce
B
Medin
Jeffrey A
JA
Platt
Jeffrey L
JL
West
Lori J
LJ
D000017:ABO Blood-Group System; D000818:Animals; D002478:Cells, Cultured; D005260:Female; D005434:Flow Cytometry; D005822:Genetic Vectors / Q000235:genetics; D015234:HLA-A Antigens / Q000378:metabolism*; D006801:Humans; D007150:Immunohistochemistry; D016086:Lentivirus / Q000235:genetics*; D008099:Liver / Q000187:drug effects; D051379:Mice; D008807:Mice, Inbred BALB C; D008954:Models, Biological; D011401:Promoter Regions, Genetic; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014161:Transduction, Genetic / Q000379:methods*
D000017:ABO Blood-Group System; D015234:HLA-A Antigens