The long-term objective of these studies is to define selected antigens of human cells using antibodies produced in non-human primates. The immunological perspective of the non-human primates should allow the recognition of epitopes on these antigens which may be difficult to be detected by lower mammalian species.
The specific aims for this renewal period are to: (1) establish the optimal conditions and method for production of chimpanzee monoclonal antibodies to selected human normal and malignant cell antigens; (2) establish the optimal conditions and methods for the in vitro transfection or infection of chimpanzee cells with oncogenes or human tumor DNA; and (3) from aim 2, establish the immunogenicity of tumorigenicity of chimpanzee cells that show evidence of gene insertion or changes in vitro growth characteristics. Specifically, aim 1 will evaluate various methods of chimpanzee B-cell fusion such as electrofusion and PEG, varying the conditions for fusion and growth as well as the parental human and murine myeloma lines to be used as fusion partners. In particular, aim 2 will evaluate various in vitro methods of gene insertion including transfection and infection of chimpanzee fibroblast, epithelial cells, or lymphocytes.
Aim 3 will utilize standard injection and serological procedures adapted in this laboratory to induce and measure the autologous immune response in chimpanzees to human gene insertion, their in vitro growth properties, and the nature of the insertion technique. (CS)
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