Isolation and characterization of an amphipathic antigen from Corynebacterium diphtheriae

Amphipathic antigen was isolated from Corynebacterium diphtheriae Park-Williams number 8 cells by extraction with 47.5% phenol, nuclease treatment and gel filtration on Sepharose 6B. The chemical composition of the amphipathic antigen was hexose (73.8%), pentose (4.6%), fatty acids (9.8%) and glycerol (4.5%). The amphipathic antigen contained arabinose and mannose as sugars at a molar ratio of 1:6 and the major fatty acids were palmitic (C16:0) acid and palmitoleic (C16:1) acid (64.2% and 26.2%, respectively). The amphipathic antigen sensitized sheep erythrocytes and had definite immunobiological activities: viz mitogenic activity on murine splenocytes, stimulatory activity on guinea pig peritoneal macrophages and human complement activation. Deacylation of the amphipathic antigen with alkali treatment lost the sheep erythrocyte-sensitizing ability and some immunobiological activities. The isolated amphipathic antigen described is not a lipoteichoic acid and is different from the teichoic acid of C. diphtheriae.