Avian antigen binding cells: enrichment methods.
Data are presented comparing different methods for the fractionation and enrichment, respectively, of specific antigen binding lymphoid cells from immunized chickens. The bovine serum albumin (BSA) anti-BSA system was chosen as a model. To enrich avian antigen binding cells (ABC) from a mixture of chicken peripheral blood and spleen lymphocytes 3 different methods were used: (1) separation of cells forming rosettes with antigen-coated sheep red blood cells (SRBC) from non-rosetting cells by density centrifugation; (2) isolation of ABC by their specific adherence to antigen bound to immunoadsorptive surfaces (gelatin, plastics); (3) column affinity chromatography with antigen-coated agarose, cross-linked dextran for plastic beads. The most efficient method was column affinity chromatography with antigen-coated polyacrylamide beads which affords up to 12-fold enrichment of ABC. Both the other methods are also suitable for separation and enrichment of specific ABC but can only with difficulty be adapted for processing the large numbers of cells which would be necessary, e.g., for in vivo transfer studies.