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Use of the Macrophage Migration Inhibition Test to Monitor Fractionation of Soluble Antigens of Chemically Induced Sarcomas of Inbred Guinea Pigs

From the Department of Microbiology and Immunology, Albert Einstein College of Medicine, Bronx, New York 10461 and the Division of Applied Immunology, Sloan Kettering Institute for Cancer Research, New York, New York

Abstract

Soluble tumor antigen was prepared from two transplanted sarcomas of strain 13 guinea pigs, induced with 3-methylcholanthrene or 9,10-dimethyl-1,2-benzanthracene, by extraction with isotonic saline followed by ultracentrifugation. Further fractionation by ammonium sulfate precipitation, gel filtration on Sephadex G-150 or Biogel A-5m, and DEAE Sephadex chromatography was performed. Antigenic activity was assayed in vivo by skin tests and in vitro by the macrophage migration inhibition method. Antigenic activity was not restricted to a single species of molecules as defined by size or charge. The major part of the activity was associated with two fractions, one excluded by Sephadex G-150, and the other included in Sephadex G-150. The behavior of the antigen preparations in isopycnic sucrose gradient centrifugation indicated that they were truly soluble and not membrane associated. The active fractions were nontoxic for the skin and for peritoneal cells and stable under the conditions of the macrophage migration test. Their antigenic activity was specific for each of the two tumors. In terms of the amount of antigen required to give a positive reaction, the migration inhibition assay was more sensitive than the skin tests.

Footnotes

¹ Supported in part by the Deutsche Forschungsgemeinschaft.

² Recipient of United States Public Health Service Career Development Award AI-19996. Supported by United States Public Health Service Grant AI-07118.

³ Supported by Grant CA-08748 of the National Cancer Institute and a grant of the Damon Runyon Memorial Fund.