The objective of this proposal is to develop techniques for the immune alteration of endocrine tissues which will allow their transplantation to allogeneic recipients without immunosuppressive therapy. To accomplish this goal, we propose studies utilizing hyperbaric oxygen culture (HOC) to modulate graft immunogenicity and antigenicity. The study consists of four interrelated parts: 1) optimization of beta cell growth and differentiation with growth factors, 2) development and optimization of HOC conditions which maintain human fetal pancreas proislet (HFP) functional viability and eliminate tissue antigenicity, 3) development of the human PBL and fetal thymus reconstituted C.B-17 scid/scid model, and 4) characterization of the allograft response to HFP proislets and immunomodulation of the response by HOC. In part 1 we will use the growth factors IGF-II, human growth hormone, and prolactin in combination with IGF-I to enhance beta cell function during human fetal pancreas (HFP) proislet culture. Cellular viability will be determined by insulin release in response to challenge in vitro and in vivo (ability to reverse experimentally-induced diabetes in nude mice). In part 2 we will correlate oxidative stress during HOC with cell viability and reduced antigenicity. We will then use oxygen free radical scavengers to prevent cellular oxidative stress while maintaining reduced antigenicity. Cell viability will be determined as described above. Altered antigenicity and immunogenicity will be determined by cellular-ELISA, immunoperoxidase staining, CML, and MLC assay in vitro. Finally, in parts 3 and 4 we will develop the human-SCID mouse chimeric model. Reconstitution extent will be determined by human antibody production, response to T and B cell mitogens, human skin graft rejection, and human DNA dot blot analysis. We will then use the human-SCID mouse model to characterize the human allograft response to HFP proislet grafts and the ability of HOC to modify the response.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
5R01DK035446-09
Application #
2139569
Study Section
Surgery, Anesthesiology and Trauma Study Section (SAT)
Project Start
1985-04-01
Project End
1995-01-31
Budget Start
1994-02-01
Budget End
1995-01-31
Support Year
9
Fiscal Year
1994
Total Cost
Indirect Cost
Name
University of Wisconsin Madison
Department
Surgery
Type
Schools of Medicine
DUNS #
161202122
City
Madison
State
WI
Country
United States
Zip Code
53715
MacKenzie, D A; Sollinger, H W; Hullett, D A (1996) Acute graft rejection of human fetal pancreas allografts using donor-specific human peripheral blood lymphocytes in the SCID mouse. Transplantation 61:1461-4
Hullett, D A; Mackenzie, D A; Sollinger, H W (1994) Successful culture of human fetal pancreas proislets in hyperbaric oxygen. Transplant Proc 26:706
Fujino, Y; Kawamura, T; Hullett, D A et al. (1994) Evaluation of cyclosporine, mycophenolate mofetil, and Brequinar sodium combination therapy on hamster-to-rat cardiac xenotransplantation. Transplantation 57:41-6
Hullett, D A; Smith, L; Sollinger, H W (1994) Enhancement of proislet functional viability with growth factors. Transplant Proc 26:709-10
Hullett, D A; Sollinger, H W (1990) Modification of allograft antigenicity. Transplant Proc 22:1926-7
Hullett, D A; Landry, A S; Sollinger, H W (1990) Mechanism of modification of allograft antigenicity. Transplant Proc 22:1928-9
Hullett, D A; Landry, A S; Leonard, D K et al. (1989) Improved human fetal pancreatic tissue survival following hyperbaric oxygen culture. Transplant Proc 21:2659-60
Hullett, D A; Bethke, K P; Landry, A S et al. (1989) Successful long-term cryopreservation and transplantation of human fetal pancreas. Diabetes 38:448-53
Hullett, D A; Landry, A S; Leonard, D K et al. (1989) Enhancement of thyroid allograft survival following organ culture. Alteration of tissue immunogenicity. Transplantation 47:24-7
Hullett, D A; Landry, A S; Sollinger, H W (1989) Regulation of class I antigens responsible for prolonged graft survival after organ culture. Transplant Proc 21:240-1

Showing the most recent 10 out of 12 publications