Antibodies produced as a result of experimental cryosurgery were shown to be highly specific with regard to the tissue frozen and the species of origin. As a consequence of destructive freezing in the coagulating gland and seminal vesicle of the rabbit, antibody is produced which reacts with extracts of the seminal vesicle, coagulating, prostate and bulbo-urethral glands, but fails to react with rabbit serum or with extracts of testis, kidney, liver, thyroid, vagina, spleen, stomach, epididymis or synovium of the rabbit. It also fails to react with prostatic or related tissue of human, guinea-pig and rat. This spectrum of tissue and species specificity, along with the proof that this antibody is an autoantibody, indicates that the same antigenic material is responsible for this antibody response as was responsible for antibodies elicited by iso-immunization by injection of tissue extracts with complete Freund's adjuvant. Additional proof was obtained of this identity by use of purified fractions of rabbit prostatic tissue. Only one of the fractions was active with an antiserum produced by iso-immunization, and this same fraction was active with an antiserum produced by cryo-immunization.
Although the same autoantigen seems to be involved, the autoantibody response to the freezing of tissue is initiated much more quickly, and rises to a maximum in a much shorter period, than the analogous autoantibody response that follows repeated intradermal injections. The cryo-immunization response is generally at a maximum within 7–10 days, whereas stimulation by injection requires a period of many weeks of repeated injections, before a similar level of antibody is reached.