Complement-Mediated Hemolysis in Agar Gel

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Abstract
Summary: Complement and rabbit anti-sheep hemolysin diffusing against each other in sheep erythrocyte-containing agar gel form two zones of hemolysis which can be related to γG and γM hemolytic antibodies. Similarly, two hemolytic zones were observed in immunoelectrophoresis experiments in which the hemolysin was electrophoretically separated in an agar medium containing sheep erythrocytes and in which lysis was induced by diffusion of complement. In many experiments, the γG hemolytic zone was separated into two areas which indicated the presence of two molecular species of γG hemolytic antibodies. This finding was confirmed by the demonstration of two antigenic types of γG antibodies separated from sheep erythrocytes. Anti-sheep hemolysins of rabbit and human origin formed circular zones of hemolysis when they were diffusing into agar containing sheep erythrocytes and complement. These zones could be reduced in size by allowing antisera with hetero- or isoantibodies for γG-globulins to diffuse against the hemolysins. The practical application of hemolysis in agar gel was explored in studying lysis of bovine erythrocytes by infectious mononucleosis sera and lysis of human erythrocytes by the hemolytic variety of human anti-A and anti-B antibodies.