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Specific Heterologous Enhancement of Immune Responses

V. Isolation of a Soluble Enhancing Factor from Supernatants of Specifically Stimulated and Allogeneically Induced Lymphoid Cells¹

From the Department of Pathology, Harvard Medical School, and the Department of Microbiology, Harvard University School of Public Health, Boston, Massachusetts 02115

Abstract

In cultures of thymocytes or spleen cells from tetanus toxoid-primed mice, the addition of 1 ng of toxoid generated the release of a soluble factor which was capable of nonspecifically enhancing the in vitro immune response of normal mouse spleen cells against sheep erythrocytes. When active supernatants were fractionated by gel filtration on Sephadex G-150 and Bio-Gel P-150, the active material eluted with molecules of approximately 75,000 m.w. Column fractions from similarly-treated supernatants from cultures of normal thymocytes or spleen cells, maintained with or without tetanus toxoid, were inactive. The enhancing activity recovered from the column fractions was degraded by treatment with the enzyme protease. A factor present in the supernatant fluid of cultures containing equal numbers of histoincompatible (allogeneic) spleen cells had similar properties. The enhancing factor was further purified by a combination of ammonium sulfate fractionation, Sephadex gel filtration, and polyacrylamide gel electrophoresis.

Footnotes

¹ This work was supported by National Institutes of Health Grants AI 05691 and AI 00221.

² Dr. Coons is a Career Investigator of the American Heart Association.