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Cytotoxic Activity of Lymphocytes

IV. Preparation of Heterologous Antibody to Human Lymphotoxin¹

From the Departments of Medical Microbiology and Surgery (Transplantation Biology), Stanford University School of Medicine, Palo Alto, California 94305

Abstract

Antibody to human lymphotoxin was elicited in rabbits immunized with partially purified supernatants of human spleen cells stimulated by phytohemagglutinin; Anti-LT activity, assayed by neutralizing soluble LT toxicity to indicator mouse L cells, is confined to the immunoglobulin fraction of the serum; it cannot be removed by adsorption to L cells, human fibroblasts or lymphocytes, or to human serum. Anti-LT activity is removed by immunoadsorption to Sepharose conjugated with goat antisera to rabbit 7S immunoglobulin; partial recovery of anti-LT activity is effected by elution with high molarity acid buffer. The amounts of antisera necessary to effect neutralization are linearly proportional to LT concentration. Anti-human LT neutralizes the toxicity of supernatants of human lymphocytes activated by other, non-PHA mitogens (antigens to which the lymphocyte donors are sensitive) and allogeneic lymphocytes in mixed leucocyte culture. However, anti-human LT is specific for human LT as it will not neutralize guinea pig or rat LT.

Footnotes

¹ This work was supported in part by National Institutes of Health Grant AM 12458 and by the Beta Sigma Phi Kidney Research Fund.