Quantitative Studies on the Action of Compound Hemolysins

Abstract

Summary A quantitative study of the characteristic curves defined in preceding papers (1, 2 and 3) led to the conclusion that the complexities of these curves as they are experimentally obtained can be removed and the curves so simplified that they assume forms which are in harmony with the views proposed by Svante Arrhenius on the reaction between hemolytic amboceptor and complement by introducing the assumption into the calculation that neither heat inactivated hemolytic amboceptor serum is free from complement nor complement serum from natural amboceptor. Geometrical methods for introducing suitable corrections were developed for this purpose. The application of such methods permits the quantitative estimation of the contaminations. Heat inactivated hemolytic amboceptor serum may, when undiluted, contain complement in quantities not much lower in order of magnitude than the complement serum. Its effect may nevertheless not be noticeable on the Wassermann reaction as long as potent amboceptor serum is used which permits a correspondingly high dilution. Application of the law of chemical mass action makes the construction of theoretical systems of characteristic curves possible. Only the exponents of the mass law equations determine the shape of these curves. Such curve systems were calculated under various assumptions regarding the chemical combination between amboceptor and complement and found to present typical pictures characteristic of each reaction. It was shown that when the corrections above referred to are applied to the curves derived from the law of chemical mass action in the opposite direction complex curves result which agree with those experimentally obtained within the limits of experimental errors except in the region of complement deviation which the theory is not designed to cover. Rational systems for expressing strength of amboceptor and complement were proposed. Conclusions were drawn regarding the relative molar concentrations of amboceptor and complement. The assumption of a stoichiometric chemical reaction between amboceptor and complement explains all observations. The stipulation of an enzymatic nature of one or both does not seem justified. Summing up the results of all four papers on this subject, we might state that while Manwaring rightfully pointed out that Arrhenius' mathematical representation of the reaction between hemolytic amboceptor, complement and red blood cells is not in agreement with the experimental facts, we find it in agreement with them so far as only the reaction between amboceptor and complement is concerned. The constants which Arrhenius used in his equations are as all constants obtained by experiment subject to revision by later investigators.