ON ANAPHYLATOXINS AND ENDOTOXINS OF THE TYPHOID BACILLUS.
The experiments recorded in this paper confirm the observations of Friedberger that acutely toxic bodies can be produced from typhoid bacilli by the action of sensitizer and complement and that, when small quantities of bacteria are used, an excess of sensitization either interferes with the formation of the poisons or leads to a cleavage of the bacterial proteid beyond the poisonous intermediate products spoken of as anaphylatoxins. Unlike the experience of other workers with poisons of this nature, however, our experiments have shown that the action of complement upon typhoid bacilli strongly sensitized or not at all sensitized may be carried on, at body temperature, for considerably longer than twelve hours without leading to a destruction of the poisons, and that this is true when the quantities of the bacteria used vary within the wide range of from one to twelve agar slants. It has been found, in fact, that in the case of this microorganism prolonged exposure at the higher temperature of considerable quantities of bacteria constitutes an unfailing method of regularly obtaining powerful poisons. The results obtained by the use of smaller quantities and the less vigorous complement action at low temperatures are far less regular or satisfactory. It would appear from this that complement action of considerable vigor is required to obtain from this bacillus any appreciable yield of anaphylatoxin, and that the poison, once formed, is not as unstable as that found in other microorganisms by Neufeld and Dold and others. In fact, although we have never observed complete lysis in vitro of the typhoid bacilli treated with antibody and complement, the sensitized bacteria exposed to the action of complement for as long as fifteen hours at 37.5 degrees C. showed, in our experiments, much disintegration, and yet powerful poisons were present. Were the influence of lysis or of the too vigorous action of the serum bodies as rapidly poison-destroying in the case of this bacillus as it has been shown to be in the case of some other bacteria, it would be hard to understand how anaphylatoxins could play any part in the toxemia of typhoid fever. This phase of our experiments, however, seems to indicate that the conditions prevailing in the infected body at the height of this disease would furnish ideal criteria for anaphylatoxin production, since, in such cases, vigorously sensitized bacilli, in large numbers, are under the prolonged influence of considerable quantities of complement, conditions exactly comparable to those prevailing in our experiments. Granted that this state of affairs is actually the case, then the subsidence of the disease might depend merely upon limitation of the supply of antigen, as the increasing bactericidal action of the blood constituents come into play, and upon the consequent diminution of the anaphylatoxin. For as the bacteria diminish and the sensitizer increases, a changed proportion between them is established which, finally, as experiment has shown, results in a failure of anaphylatoxin production. For although our experiments have shown that, within a wide latitude of relative proportions of bacteria and antibody, anaphylatoxin can be formed, beyond this range an excess of one or the other element eventually will prevent their formation. It is not, however, the purpose of this paper to discuss the mechanism of the subsidence of the disease since this phase of the work will necessitate further experimental study. In regard to the experiments with kaolin, we were unable to confirm the contention of Keysser and Wassermann, though it is more than likely that toxic bodies could be formed by the action of complement upon any foreign proteid rendered amenable to its action. We are not inclined to attribute too much importance to these negative results, recording them merely as they occurred. However, should it be found subsequently that anaphylatoxins can be formed in this way, it seems unlikely that they are formed from the sensitizer or amboceptor as matrix, since this was not specifically adsorbed out of concentrated serum by the kaolin in our experiments. On the basis of experiments with so called endotoxins, ,we feel that the existence of such preformed intracellular poisons as an element in typhoid toxemia has not been proved, and is not absolutely necessary for the explanation of the phenomena occurring in this disease. However, the diarrhea, the hemorrhagic lesions, and the protracted symptoms following the injection of extracts and filtrates of the bacillus, differing so strikingly from the acute illness with rapid death or equally rapid recovery resulting from anaphylatoxin poisoning, would justify the assumption that poisons of this nature may still play a part in the disease, adding an additional specific characteristic to the clinical picture. As stated before, however, it is not improbable that all these characteristics may represent merely a more protracted or subacute state of anaphylatoxin toxemia. The experiments with autolysates, although none of them were fatal in their results upon guinea pigs, have sufficiently indicated that poisons comparable to anaphylatoxins can be formed in this way. This would indicate that a reaction of proteolysis, which may take place slowly by autolysis, is hastened by the action of complement, and its velocity is still further augmented by the increase, within certain limits, of the sensitization,-a conception which would attribute to the combined action of complement and sensitizer a function not incomparable to that of the bodies spoken of as catalytic agents.
Division of Bacteriology, Department of Medicine, Stanford University, California.
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Zinsser