Despite recent successes, a major impediment to progress in clinical transplantation continues to be allograft rejection and morbidity and mortality resulting from non-specific immunosuppression. Therefore, achievement of long lasting donor-specific unresponsiveness to foreign antigens would constitute an important clinical advance. This FIRST proposal will examine the potential use of gene transfer technology to induce antigen-specific transplantation tolerance. More precisely, these studies will determine whether tolerance can be induced in lethally irradiated mice reconstituted with autologous bone marrow cells into which xenogeneic MHC genes have been introduced and expressed. Retroviral expression vectors, which contain the human HLA-A2 gene will be constructed and introduced into packaging cell lines. Colonies which produce high titer, replication-defective recombinant retroviruses will then be co-cultured with mouse bone marrow cells to attain optimal gene transfer. Irradiated C57B1/6 (B6) mice will then be transplanted with autologous bone cells transformed to express the human HLA-A2 gene. Long-term bone marrow reconstituted B6 mice will then be examined for immunologic reactivity to the HLA-A2 antigen by 1) in vitro immunologic assays using lymphocytes for stimulation from cell lines or transgenic mice expressing the HLA-A2 gene on the B6 background and 2) by transplantation of skin grafts from HLA-A2 transgenic or third-party control donor mice onto reconstituted B6 recipients. The successful completion of these experiments should provide insight into xenogeneic tolerance induction and will provide an impetus to further work designed to study tolerance across more complicated histocompatibility barriers for clinical application.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI031641-07
Application #
6169676
Study Section
Surgery, Anesthesiology and Trauma Study Section (SAT)
Program Officer
Kerr, Lawrence D
Project Start
1993-01-01
Project End
2003-05-31
Budget Start
2000-06-01
Budget End
2001-05-31
Support Year
7
Fiscal Year
2000
Total Cost
$281,070
Indirect Cost
Name
University of Nebraska Medical Center
Department
Surgery
Type
Schools of Medicine
DUNS #
City
Omaha
State
NE
Country
United States
Zip Code
68198
Mashalova, Elena V; Guha, Chandan; Roy-Chowdhury, Namita et al. (2007) Prevention of hepatocyte allograft rejection in rats by transferring adenoviral early region 3 genes into donor cells. Hepatology 45:755-66
Zhang, Jia Lin; Cai, Jin; Walls, Shannon et al. (2005) Tolerance by selective in vivo expansion of foreign major histocompatibility complex-transduced autologous bone marrow. Transplantation 80:362-9