Production of Shwartzman Reaction in Rabbits with Purified Fraction of B. coli Filtrate

Previous publications 1 , 2 from this laboratory described a method for the separation and concentration of the factor in B. coli filtrates which produces hemorrhage in mouse tumors. Since B. coli filtrates are known to produce the Shwartzman reaction in rabbits, it was of interest to ascertain whether the purest fraction of the tumoraffecting agent obtained thus far 2 was also capable of eliciting hemorrhage in prepared sites in the skin of rabbits. It was also desirable to make quantitative comparisons between the original filtrate and the purified fraction as to their ability to produce hemorrhage in tumors of mice as well as the Shwartzman reaction in the rabbit.

Equivalent tumor-affecting doses were determined by performing a preliminary titration in mice bearing cutaneous tumors, according to a technic described elsewhere. 3 Strain M mice bearing 6-day-old cutaneous growths of sarcoma 37 were given intra-abdominal injections of diluted filtrate or dilutions of the purified fraction. All dilutions were made with 0.9% NaCl solution. It was found that 0.2 cc. of a 1:50 dilution of the original filtrate produced hemorrhage in tumors, but 0.2 cc. of a 1:100 dilution failed to do so. The purified fraction was active down to 0.2 cc. of a 1:1500 dilution of a solution which contained 2 mg. of the purified fraction per cc. Thus the minimal tumor-affecting dose of this purified fraction was 0.26γ.

Comparable doses of the two materials were then injected into rabbits according to Shwartzman's technic. The endermal injections consisted of 0.2 cc. doses. Each of 4 rabbits received preparatory injections of 3 dilutions of the original filtrate and of 3 dilutions of the purified fraction, as shown in Table I. Twenty-four hours later 2 of the rabbits each received 0.5 cc. of undiluted filtrate intravenously while the other 2 received 0.5 cc. of a 1 :10 dilution of the purified fraction. The results are shown in Table I.