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Cell Separation on Affinity Columns: The Preparation of Pure Populations of Anti-Hapten Specific Lymphocytes¹

From the Department of Bacteriology and Immunology, University of California, Berkeley, California, and the Service de Immunologie Cellulaire, Institut Pasteur, Paris, France

Abstract

Procedures were developed for purifying anti-hapten specific lymphocytes on an immunoadsorbent. Mouse spleen cells were filtered through large polyacrylamide beads with covalently attached -lactoside hapten groups. Specifically binding cells were eluted with hapten from washed affinity beads. Hapten-eluted spleen cell populations derived from non-immune mice were much smaller in number than those from mice immunized against hemocyanin azophenyl--lactoside. Pure populations of anti-hapten specific cells from immune spleens were greatly enriched in indirect plaque-forming cells against -lactoside-coupled sheep red blood cells.

Footnotes

¹ This work was supported by Grant AI-06610 from the National Institute of Allergy and Infectious Diseases, United States Public Health Service.

² Present address: Service de Microbienne Cellulaire, Institut Pasteur, Paris 15, France.