Studies on the Virulence of Tubercle Bacilli

The bacillary cords characteristic for virulent tubercle bacilli are readily disrupted when wet bacilli are suspended in hydrocarbons such as paraffin oil or petroleum ether. The disruption of cords is due to the removal of a material coating the surface of the bacilli and causing them to adhere to each other. This material can be obtained from virulent bacilli by extracting them with petroleum ether. It is a lipid. Avirulent variants of tubercle bacilli do not yield a similar material after extraction in the same manner; only little of it is obtained from BCG bacilli. The following properties of the fraction obtained by petroleum ether extraction are described: (a) It inhibits the migration of leukocytes in vitro. (b) If repeatedly injected in small doses into mice, it is toxic, whereas a single high dose does not give rise to toxic manifestations. (c) The susceptibility of mice to the toxic action of repeated injections parallels to some extent their degree of susceptibility to infection with the strain of tubercle bacilli from which the fraction was obtained. (d) The injection of the extracted material into guinea pigs does not induce a state of allergic reactivity toward tuberculin. Likewise, tuberculin-positive guinea pigs do not show hypersensitivity against injections of the extracted substance. Bacilli extracted with petroleum ether do not lose their viability. They grow out normally in vitro, and they are still pathogenic. However, the removal of the petroleum ether-soluble lipid from the bacilli results in a loss of the ability of the organisms to inhibit the migration of polymorphonuclear leukocytes. Moreover, mice and guinea pigs infected with extracted bacilli may develop tuberculosis considerably slower than animals injected with comparable amounts of unextracted organisms. The significance of these findings is discussed in relation to the problem of the virulence of tubercle bacilli.