Studies of Brucella lipopolysaccharide

Brucella endotoxin differs from other gramnegative endotoxins in that it is recovered in the phenol phase rather than the aqueous phase of the Westphal hot phenol water procedure. This was the first described from this laboratory by Redfearn (1960) with phenol-killed smooth B. abortus and B. melitensis and has since been confirmed by others. Preliminary extraction of brucella cells with acetone, as called for in the original Westphal procedure, was followed by Renoux et al. (1973) who reported that the aqueous phase lacked endotoxic activity and the phenol phase had very low toxicity. In order to test the hypothesis that prior acetone extraction removes lipid A, we have repeated the Redfearn procedure with acetone extracted cells and have confirmed that the major portion of the endotoxic activity resides in the phenol phase. Acetone treatment does not remove the lipid A believed to be responsible for mouse lethality as well as necrotizing activity in guinea pig and rabbit skin. Preparations of brucella endotoxin (lipopolysaccharide) contain varying amounts of polypeptide some of which is tightly bound. The dermal response of sensitized guinea pigs to brucella LPS was shown to be a combination of reactions comprising those due to (1) innate toxicity of lipid A, (2) antibody mediated reactions due to polysaccharide portion of the molecule and (3) delayed hypersensitivity due to polypeptide portion of the molecule.