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Cluster Formation in Vitro by Mouse Spleen Cells and Sheep Erythrocytes¹

Department of Experimental Biology, Roswell Park Memorial Institute, Buffalo, New York 14203

Abstract

The primary injection of sheep erythrocytes into young adult mice was followed by the appearance in the spleen of distinct populations of immunocytes secreting specific antibody. Different types of immunocytes were recognized by the properties of the antibody produced, i.e., the ability to lyse erythrocytes in agar gel directly and indirectly upon facilitation (plaque-forming cells), and the ability to agglutinate erythrocytes in liquid medium (cluster-forming cells). The optimal conditions for enumeration of cluster-forming cells were established. The number of cluster-forming cells in the spleens was much greater than that of plaque-forming cells, and the kinetics of appearance were different. Furthermore, previous cluster formation in liquid suspension did not influence the subsequent appearance of hemolytic plaques in agar gel by the same spleen cell preparation. The plaques developed around immunocytes other than the cluster-forming cells. It was concluded that cluster-forming cells and plaque-forming cells are engaged in the exclusive production of hemagglutinins and hemolysins, respectively.

Footnotes

This investigation was supported in part by Grant G-66-RP-5 of the United Health Foundation of Western New York and by Institutional Grant IN-54 of the American Cancer Society.

² Submitted in partial fulfillment of requirements for the Ph.D. degree from the Institute of Radiation Biology, the University of Tennessee, Knoxville, Tennessee. Postdoctoral Fellowship (1968) of the Damon Runyon Memorial Fund for Cancer Research.