Studies on the gonococcus

Abstract

Serologically active substances have been isolated from the gonococcus as well as from the broth in which the organism has been grown, but as yet not all of them have been chemically defined. Several investigators (DeChristmas, 1897;Herrold, 1927;Clark, Ferry and Steele, 1931; and Wolifenstein and Pieper, 1931) have reported that broth in which the gonococcus has been grown elicits a typical dermal reaction in patients with gonococcal infection but not in persons without evidence of the disease. These findings have been interpreted as evidence that the gonococcus elaborates an immunologically active substance during growth in a broth medium. Recently, Rossett (1939), working with gonococcal bouillon filtrates, separated a fraction which possessed antigenic properties similar to those noted by previous workers. He considered this material to be a carbohydrate, a conclusion which was not supported by quantitative chemical proof.By fractionation procedures similar in a number of respects to those employed by Rossett, we have isolated a fraction from gonococcal broth supernatants which was antigenic and toxic for laboratory animals. The present report describes the isolation and properties of this fraction, which is proteic in nature. No evidence was found to indicate that we were dealing primarily with a carbohydrate.MATERIALS AND METHODS Preparation offraction Seven strains of Neisseria gonorrhoeae, which had been under artificial cultivation for from 1 month to 34 years, were selected for the study. The organisms were grown in 300-ml. flasks containing 100 ml. of Douglas's broth to which 0.5 per cent each of sodium phosphate and sodium nitrate had been added. After 3 days of incubation at 37°C. the cells were removed by centrifugation. The broth supernatants were concentrated ten-fold in vacuo at temperatures not exceeding 20°C., and the broth constituents of low molecular weight were then separated by dialysis in the cold. Small amounts of nucleoprotein were removed from the broth concentrate by precipitation at pH 4.5. The resulting supernatant, when brought to neutrality and dialyzed against 25-per cent alcohol I These studies were supported by a grant from the John andMary R. Markle Foundation. 2 This paper was presented in preliminary form at the 43rd Annual Meeting of the Society of American Bacteriologists,

The Chemistry and Metabolism of Bacteria

“…Stockinger, Ackerman & Carpenter, for example, reported extensive studies of the composition of the gonococcus (12) and its products (13). They described two nucleoprotein fractions obtained by extraction of mass cultures.…”