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Lymphocyte Plasma Membranes

V. Specificity and Mitogenicity of Absorbed anti-Lymphocyte Sera¹

From the Department of Immunology in the Research Institute of The Hospital for Sick Children at the University of Toronto, Toronto, Ontario, Canada

Abstract

Anti-lymphocyte sera against human thymus [ALS-(THY)] were absorbed serially with cultured human lymphoblasts (CHL) or thymus and residual antigen-binding activity was tested. The absorbed ALS were used to bind ¹²⁵I-labeled antigens from lymphocytes labeled by the lactoperoxidase catalyzed iodination technique. Absorption of ALS(THY) with CHL led to the absorbed serum having less than 5 to 10% of its original antigen-binding activity against labeled CHL antigens while maintaining from 20 to 40% of its original activity against labeled THY. Serial absorption of ALS(THY) with THY led to an equal decrease in activity against both THY and CHI. When the immunoprecipitates from these experiments were examined on polyacrylamide gels containing SDS it was found that serial absorption of ALS(THY) with THY first removed activity against a component of m.w. 48,000 leaving relatively greater activity against material of apparent high molecular weight. In contrast, absorption of ALS(THY) with CHL removed the antibodies against the high molecular weight material while leaving activity against the component of m.w. 48,000. When these absorbed ALS were used to induce in vitro lymphocyte proliferation, it was found that ALS(THY) absorbed with human THY lost its mitogenicity whereas ALS(THY) absorbed with CHL, did not. The retention or loss of mitogenicity seemed to correlate with retention or loss of binding activity against the component(s) of m.w. 48,000.

Footnotes

¹ This work was supported by Medical Research Council of Canada.

² Dr. Zimmerman is a scholar of the Medical Research Council.