Class II Human Leukocyte Antigens (HLA) regulate the overall immune response and also trigger the rejection of a transplanted organ.
The specific aim of the proposed research is to downregulate the expression of HLA class II antigens (HLA) . The inhibition of expression of HLA class II genes will be achieved by transfecting cells with the recombinant molecules containing the cloned cDNA from the D region of the HLA complex in antisense orientation. These constructs will have the dominant selectable markers, hph or neo for selection or the mutant dihydrofolate reductase (DHFR*) gene for selection and amplification of the transgenomic fragments in mammalian cells. Expression vectors, episomal replicons and retroviral vectors will be used for the construction of the recombinant molecules. The molecules containing the insert will be transferred into the cells by electroporation in case of the expression vectors and episomal vectors and by infection when the retrovirus vectors are used as the means of gene transfer. Reduction in cell surface expression of class II antigens on the drug resistant cell lines will be evaluated by monoclonal antibody staining with flow cytometric determinations and by the ability to activate cell proliferation in the mixed lymphocyte culture. The long-term goal of the proposed project is to apply the technique of antisense cDNA to the downregulation of the expression of HLA class II antigens in bone marrow cells and to induce tolerance to transplantation of mismatched and haplocompatible bone marrow stem cells to the recipient. Non-human primates will be used for preclinical trials.

Project Start
Project End
Budget Start
Budget End
Support Year
5
Fiscal Year
1994
Total Cost
Indirect Cost
Name
University of California San Francisco
Department
Type
DUNS #
073133571
City
San Francisco
State
CA
Country
United States
Zip Code
94143
Cowan, M J; Chou, S H; Tarantal, A F (2001) Tolerance induction post in utero stem cell transplantation. Ernst Schering Res Found Workshop :145-71
Carrier, E; Lee, T H; Busch, M P et al. (1997) Recruitment of engrafted donor cells postnatally into the blood with cytokines after in utero transplantation in mice. Transplantation 64:627-33
Mychaliska, G B; Rice, H E; Tarantal, A F et al. (1997) In utero hematopoietic stem cell transplants prolong survival of postnatal kidney transplantation in monkeys. J Pediatr Surg 32:976-81
Kline, R M; Stiehm, E R; Cowan, M J (1996) Bone marrow 'boosts' following T cell-depleted haploidentical bone marrow transplantation. Bone Marrow Transplant 17:543-8
Sorof, J M; Koerper, M A; Portale, A A et al. (1995) Renal transplantation without chronic immunosuppression after T cell-depleted, HLA-mismatched bone marrow transplantation. Transplantation 59:1633-5
Carrier, E; Lee, T H; Busch, M P et al. (1995) Induction of tolerance in nondefective mice after in utero transplantation of major histocompatibility complex-mismatched fetal hematopoietic stem cells. Blood 86:4681-90
Elder, M E; Hope, T J; Parslow, T G et al. (1995) Severe combined immunodeficiency with absence of peripheral blood CD8+ T cells due to ZAP-70 deficiency. Cell Immunol 165:110-7
Cowan, M J; Golbus, M (1994) In utero hematopoietic stem cell transplants for inherited diseases. Am J Pediatr Hematol Oncol 16:35-42
Dror, Y; Gallagher, R; Wara, D W et al. (1993) Immune reconstitution in severe combined immunodeficiency disease after lectin-treated, T-cell-depleted haplocompatible bone marrow transplantation. Blood 81:2021-30