SEROLOGICALLY DETECTABLE RABBIT HISTOCOMPATIBILITY ANTIGENS SOLUBILIZED BY SEVERAL AGENTS

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Abstract
Three water-soluble preparations were obtained from cell membrane fragments of rabbit spleen and lymph nodes and give serological evidence of containing rabbit histocompatibility antigens. These materials were solubilized by the use of Triton X-100, and by snake venom phos-pholipase and butanol from the water-insoluble fraction of the Triton extract. The evidence for soluble histocompatibility antigens in these preparations is given by their reaction with antibodies present in rabbit antiserum to rabbit leucocytes, as indicated by their ability to agglutinate and tanned sheep red blood cells (RBC) coated with specifically purified antibody from the antileucocyte serum, by their ability to render tanned RBC agglutinable by the antileucocyte serum and by their inhibition of the agglutination of antigen-coated RBC by antileucocyte serum. The butanol-solubilized material has about 12 times the specific activity of the phospholipase-solubillzed material; both yield approximately the same total amount of soluble antigens. These 2 materials are entirely excluded from Sephadex G-200, whereas the greater part of the Triton-solubllized is not excluded. On diethylaminoethyl [DEAE] cellulose chromatography of the butanol-solubilized preparation, almost all of the serological activity can be found within approximately 1/2 the total protein applied to the column.