Adaptation of the Hemolytic Plaque Technique for Enumeration of Immune Cells Responding to Heterologous Immunoglobulin Antigens

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Abstract
The cytodyna-mics of the primary immune response of rabbits injected with heterologous mouse immunoglobulins was studied by means of the hemolytic plaque technique. Using indicator sheep red blood cells (SRBC) coated with sub-hemolytic amounts of mouse anti-SRBC serum and spleen cells from the immunized rabbits, it was possible to detect and enumerate single antibody producing cells. Although the magnitude and timing of the cellular responses were variable, peak was attained by the 7th or 8th day and had completely subsided by the 15th day. The serologic response, which was followed by passive hemagglutination, closely paralleled the cellular response. The absence of spontaneous background distinguished protein reactive plaque-forming cells from those reactive against erythrocyte antigens.