Tumour-associated antigens reacting with cytotoxic antibodies in serum of hepatoma-bearing rats.
3M-KCl extracts of the hepatoma D23 contain antigens that inhibit the complement-dependent cytotoxicity for D23 hepatoma cells of serum from D23 tumour-bearing rats (D23 TBS). Inhibition was not due to a general anticomplementary activity of the extracts. Although a minor part (25%) of the protein of D23-KCl extract was insoluble in PBS, this part contained most of the inhibitory activity. Fractionation of the PBS-soluble material of the extract on Concanavalin A-Sepharose showed that the inhibitory activity did not bind to the lectin. Analysis of D23-KCl extracts on a Sepharose CL-4B column showed that the antigens involved in the cytotoxicity were heterogeneously distributed in the high-mol. wt region (greater than 200,000). Precipitation with 10% trichloroacetic acid (TCA) of D23 KCl extracts revealed that most of the antigenicity was insoluble in TCA. Heating of D23 KCl extracts at 100 degrees C did not affect the antigenicity. Enzyme treatment of D23 extra nuclear membranes (D23 ENP) revealed that the inhibitory activity was not sensitive to proteolytic digestion, while treatment with phospholipase A2, C or D abrogated partly the inhibitory activity. The lipid nature of the antigenicity was indicated by its solubility in organic solvents as chloroform or n-butanol.
D000818:Animals; D000912:Antibodies, Neoplasm / Q000032:analysis*; D000951:Antigens, Neoplasm / Q000276:immunology*; D002846:Chromatography, Affinity; D003165:Complement System Proteins / Q000276:immunology; D003602:Cytotoxicity, Immunologic; D004591:Electrophoresis, Polyacrylamide Gel; D008114:Liver Neoplasms, Experimental / Q000276:immunology*; D008970:Molecular Weight; D011506:Proteins / Q000032:analysis; D051381:Rats; D011919:Rats, Inbred Strains
D000912:Antibodies, Neoplasm; D000951:Antigens, Neoplasm; D011506:Proteins; D003165:Complement System Proteins