Human Hybridomas for Exotic Antigens.
Abstract
We have investigated methods for in vitro induction of B cells from human peripheral blood and spleen using a prototype exotic antigen Keyhole limpet hemocyanin and for reference a prototype conventional antigen Diphtheria toxoid. To ensure that maximal induction would occur, various sources of growth and differentiation factors were added to cultures. An enhanced ELISA assay capable of detecting 1-10 ng/ml of specific antibody was employed, allowing us to detect the antibody secreted by 20-200 cells in a culture volume of 0.2 ml antibody-producing cells secrete sufficient antibody in one day to reach 10 ng in one ml-or. We also attempted to capture induced B cells at an early stage of fusion with the human tumor cell line WI-L2-729-HF2. Despite extensive testing we were unable to reliably detects specific antibody secretion by any of the methods tested. In conclusion we draw attention to the strong possibility that the immune system (whether in mice or humans) is structured in such a way that primary in vitro immune responses will be impractical due to the low frequency of inducible antigen-specific B cells in both peripheral blood lymphoid organs. Keywords: Monoclonal antibodies. (aw)
Document Details
- Document Type
- Technical Report
- Publication Date
- Jan 01, 1989
- Accession Number
- ADA207481
Entities
People
- Melvin Cohn
Organizations
- Salk Institute for Biological Studies