Enrichment and expansion of specific antibody-forming cells by adoptive transfer and clustering, and their use in hybridoma production.
Adoptive transfer regimens have been examined as a method of enriching and expanding antibody-forming cells (AFC). When spleens from mice which had reverted to memory or from those at the peak of an AFC response were transferred to syngeneic irradiated recipients, a comparable enrichment in AFC of about 10-fold was found. However, recently re-stimulated spleen cells gave much better expansion of total AFC in the recipient mice. The degree of expansion was examined using different routes and timing of antigen stimulus and AFC recovery. With the optimum protocol found the AFC pool obtained from adoptively-transferred recipients was on average 80-fold greater than from conventionally re-immunised mice in a number of experiments. Further enrichment of the AFC was shown by an in vitro clustering technique which gave suspensions with AFC enriched to better than 1 cell in 10. Cluster-enriched and adoptive-transfer enriched populations were both shown to give a much higher incidence of successful specific hybridoma production than spleen cells from conventionally re-immunised mice.
D000818:Animals; D000917:Antibody Formation; D002459:Cell Fusion; D005260:Female; D006462:Hemolytic Plaque Technique; D006825:Hybridomas / Q000276:immunology*; D007156:Immunologic Memory; D008214:Lymphocytes / Q000276:immunology; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D008808:Mice, Inbred CBA