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Studies on Immunity in Anthrax

III. ELABORATION OF PROTECTIVE ANTIGEN IN A CHEMICALLY-DEFINED, NON-PROTEIN MEDIUM

From the Chemical Corps Biological Laboratories, Camp Detrick, Frederick, Maryland

Abstract

Elaboration of the protective antigen of Bacillus anthracis was obtained in a chemically-defined, non-protein medium composed of 17 amino acids, inorganic salts, adenine, guanine, uracil, thiamine, glutamine, glucose, and sodium bicarbonate. Sterile filtrates of static 20-hour cultures of a non-proteolytic mutant strain of B. anthracis, when injected in divided doses totaling 1 ml., were capable of immunizing rabbits so that they survived intracutaneous challenge with 10,000 spores of virulent B. anthracis, representing at least 100 lethal doses. The preparations immunized guinea pigs against a challenge of 1000 spores and, when precipitated with alum, produced detectable immunity in mice. A preliminary experiment suggested that the antigen also produced effective immunity in monkeys. Tentative indications as to some of the critical constituents and cultural conditions for production of antigen were obtained. Protective antigen activity of the crude culture filtrate was relatively unstable; the protective antigen could, however, be concentrated and stabilized by lyophilization or by alum precipitation. The meaning of the results is discussed.