Antilisterial immunity includes specificity to listeriolysin O (LLO) and non-LLO-derived determinants.

Subclinical infection of BALB/c mice with virulent Listeria monocytogenes leads to the generation of Listeria-specific T-cell populations required for the expression of protective immunity. The L. monocytogenes-produced hemolysin listeriolysin O (LLO) is a virulence factor which appears to be crucial for the induction of protective antilisterial immunity. Analysis of the specificity of antilisterial cytotoxic cells from Listeria-immune BALB/c donors has shown a dominant response to an epitope corresponding to amino acids 91 to 99 of LLO. Demonstration of antilisterial T cells with specificity to non-LLO-derived epitopes has been difficult to achieve because of the requirement of LLO in facilitating escape of the bacteria to the cytoplasm of the host cell and the apparent dominance of an anti-LLO response in antilisterial immunity. In this study we show that antilisterial immunity also includes specificity to non-LLO-derived determinants. We used as an immunogen an LLO- mutant of L. monocytogenes which expresses the hemolysin perfringolysin O (PFO). The LLO- PFO+ L. monocytogenes mutant possesses invasive properties similar to those of wild-type L. monocytogenes and escape from the phagocytic vacuole because of the activity of PFO. We found that J774 target cells infected with the LLO- PFO+ L. monocytogenes mutant were lysed by antilisterial cytotoxic T cells obtained from BALB/c mice immunized with wild-type L. monocytogenes. In addition, BALB/c mice immunized with the LLO- PFO+ L. monocytogenes mutant were immune to challenge with LLO+ wild-type L. monocytogenes, a finding indicative of protective antilisterial immunity specific to Listeria-derived epitopes other than LLO. Spleen cells from BALB/c mice immunized with the LLO- PFO+ L. monocytogenes mutant adoptively transferred antilisterial protection to a subsequent challenge with wild-type L. monocytogenes. This splenocyte population also contained cytotoxic cells which lysed target cells infected with either the LLO- PFO+ L. monocytogenes mutant or wild-type LLO+ L. monocytogenes but did not lyse target cells infected with an LLO-expressing Bacillus subtilis transformant. These results establish that during the immune response to L. monocytogenes, immune splenocytes with specificity for LLO and other, non-LLO-derived epitopes develop. These non-LLO epitopes serve as targets for antilisterial cytotoxic cells and for lymphocytes which adoptively transfer antilisterial immunity.