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The Journal of Immunology, 1976, 116: 1324-1331.
Copyright © 1976 by The American Association of Immunologists, Inc.
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Naturally Soluble Tumor Antigens from Guinea Pig Hepatomas: Isolation and Partial Characterization

Barbara Detrick-Hooks, Howard G. Smith, Robert C. Bast, Jr.1, Virginia C. Dunkel2 and Tibor Borsos

From the Biology Branch, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20014

Abstract

Naturally soluble tumor antigens were detected in the ascites fluid of guinea pigs bearing an ascites tumor and from exhausted tissue culture media of cultured tumor cells. Two antigenically distinct cell lines of diethylnitrosamine-induced strain-2 guinea pig hepatomas (line-10 and line-1) served as the source of tumor antigens. Tumor antigen activity was detected by four different techniques: immunodiffusion, inhibition of complement-mediated cytotoxicity, inhibition of membrane immunofluorescence, and delayed cutaneous hypersensitivity. With syngeneic tumorspecific antiserum, line-10 guinea pig tumor antigens were detected by immunofluorescence in the concentrated ascites and tissue culture fluids. With a xenogeneic antiserum, demonstrated to be tumor specific, line-10 tumor antigens were detected not only in the concentrated ascites and tissue culture fluids but also in two of the partially purified fractions of these fluids. When the line-10 concentrated ascites and its fraction I were subjected to ultracentrifugation at 300,000 x G for 1 hr, the antigen activity was retained in the supernatant and thus by this criterion the tumor antigens detected in these samples are soluble.

Immunodiffusion data indicate that more than one antigen is present in the line-10 system since three lines of precipitation were detected when line-10 concentrated ascites was reacted with the line-10 tumor-specific antiserum. In contrast to this, the line-10-concentrated tissue culture fluid displayed only one line of precipitation.

Although tumor antigens could not be demonstrated in the other antigenically distinct tumor cell line, line-1, by immunodiffusion or inhibition of membrane immunofluorescence, inhibition of complement-mediated cytotoxicity was able to detect tumor antigens in the line-1 concentrated ascites and tissue culture fluids.

Footnotes

1 Present address: Peter Bent Brigham Hospital, Boston, Massachusetts 02115.

2 Present address: Office of the Associate Director, Carcinogenesis Program, DCCP, NCI, Bethesda, Maryland 20014.






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