The resistance of white mice to tuberculous infection could be increased by preliminary vaccination with small amounts of tubercle bacilli killed by contact with 2 per cent phenol. Vaccine prepared from a variant strain of human tubercle bacilli unable to multiply in vivo (H37Ra) proved as active as vaccines prepared from either virulent or attenuated strains.
The immunity induced by phenol-killed bacilli persisted for several weeks. Under the conditions of the experiments, however, it was never able to bring about the death of the virulent bacilli used for the challenge infection, even when the infective inoculum was very small. Its protective effect could be detected (a) by the increased survival time of mice infected with a very large dose of virulent bacilli, and particularly (b) by the lower numbers of bacilli present in the organs of mice sacrificed at various periods of time after injection of sublethal infective doses. Under the proper conditions of vaccination the immunity produced in mice by phenol-killed cells of avirulent bacilli was of the same order as that produced by BCG.
The protective antigen proved to be susceptible to heat, particularly at acid reactions. It retained its activity when the bacilli were disintegrated and rendered non-acid-fast by grinding with concentrated phenol. It remained in the insoluble cellular debris when the bacilli were extracted with 88 per cent phenol.
Reasons are presented to support the view that the antigenic components present in the tubercle bacilli (avirulent as well as virulent) killed with phenol play a significant part in several manifestations of increased resistance to tuberculosis.