He showed clearly that the first injection of an animal toxin sensitized the test animal to even a very small second injection, and that, with very small doses, the violent symptoms following the second injection were out of all proportion to the mild symptoms following the first. He also established that, to produce the violent reaction, there must be an interval of several weeks between the injections.
In 1903 Nicolas Maurice Arthus of Lausanne described the Arthus phenomenon. If a rabbit was injected subcutaneously with repeated doses of horse serum, no effect was produced by the subsequent injections at first, but as the interval from the first injection lengthened, the injection site became swollen, hardened, and ulcerous. In 1905 Richard Otto showed that it was not the toxin in the "diphtheria antitoxin" (at that time a mixture of toxin and antitoxin was used) that produced serious effects in guinea pigs injected with repeated small doses at long intervals, but the horse serum in which the toxin was contained. Further, the reaction depended not upon the dose but upon the time interval. It was soon shown that a guinea pig injected with horse serum showed no hypersensitivity to the serum of other animals, and also that specific reactions occurred after the injection of milk, egg, or muscle extract. It was thus conclusively demonstrated that Richet's anaphylaxis was due to the injection of any protein, whether or not it was toxic on the first injection.
In 1907 Richet showed that, if the serum of an anaphylactic dog was injected into a normal dog, the latter became anaphylactic. The anaphylactic state could therefore be passively transmitted, and it was an antigen-antibody reaction. He continued to study anaphylactic phenomena, and for his work he was awarded the Nobel Prize in 1913. Anaphylaxis is closely associated with serum sickness and allergy, and later investigations of allergic diseases stem from Richet
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