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Factors Involved in the Cytotoxicity of Normal Guinea Pig Serum for Cells of Murine Tumor TA3 Sublines Treated with Neuraminidase

From the National Institute for Medical Research, Mill Hill, London NW7 1AA, England, and the Department of Pathology, Massachusetts General Hospital, Boston, Massachusetts 02114

Abstract

Treatment of two ascites sublines of the TA3 tumor cell with neuraminidase under conditions that removed 70 to 75% of the total cell surface sialic acid exposes receptors on the cell surface that react with cytotoxins in normal serum from unsensitized guinea pigs. Removal of comparable amounts of cell surface sialic acid from TA3 cells by trypsin in the form of sialoglycopeptides did not render the cells susceptible to lysis by the guinea pig serum. Treatment of neuraminidase-treated cells with trypsin under identical conditions reduced the susceptibility of the cells to lysis by only 35 to 40%. The requirement for neuraminidase appears to lie in the hydrolysis of glycoside linkages between sialic acid residues and penultimate sugars in cell surface glycoproteins and possibly glycolipids, with exposure of new terminal oligosaccharide sequences. Simple changes in the net surface charge or membrane fluidity of TA3 cells brought about by neuraminidase treatment were considered less likely to be responsible for the increased susceptibility of the treated cells to lysis. Hapten inhibition studies on the cytotoxic effects of normal guinea pig serum on neuraminidase-treated TA3 cells using glycoproteins with defined terminal oligosaccharide sequences have in general been unconvincing. However, glycoproteins containing terminal -galactosyl residues showed weak but definite inhibitory activity. The presence of such residues at the surface of both sublines of the TA3 tumor cell was demonstrated by agglutination of the cells with a galactose-binding lectin from Ricinus communis.

The cytotoxicity of the normal guinea pig serum against neuraminidase-treated TA3 cells was concentrated in the 19S IgM fractions of guinea pig immunoglobulins and was labile to mild heat treatment and during storage at 4°C. No agglutination of neuraminidase-treated TA3 was found with guinea pig serum either at 37° or 4°C. The role of similar cytotoxins in C3H mouse serum in the transplantation properties of the strain-specific TA3-St and the nonspecific subline TA3-Ha and derivatives derived from them by neuraminidase treatment are briefly discussed.

Footnotes

¹ Present address: Department of Microbiology, Harvard School of Public Health, Boston, Massachusetts 02115.

² These authors' work is supported by United States Public Health Service Grant CA 14922 from the National Cancer Institute.

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