An in silico DNA vaccine against Listeria monocytogenes.
The present paper analyzes the cytokine response of mouse macrophages during infection by Listeria monocytogenes. The use of different mutants of L. monocytogenes impaired in various steps of the infection process allowed us to dissect the cytokine response. Cytokine mRNA expression was detected by PCR-assisted amplification of RNA extracted from macrophages after infection with different Listeria strains. An increase in the amount of mRNA for tumor necrosis factor alpha (TNF-alpha), interleukin-1 alpha (IL-1 alpha), IL-1 beta, and IL-6 was detected in P388D1 macrophages infected with L. monocytogenes at 4 h postinfection. Interestingly, only hemolytic strains of L. monocytogenes were able to induce IL-1 alpha, IL-6, and TNF-alpha mRNA. This indicated that the induction of these cytokine mRNAs requires entry of the listeriae into the host cell cytoplasm. In contrast, IL-1 beta was also induced by infection with nonhemolytic mutants of L. monocytogenes which remain entrapped within the phagosome. The levels of TNF, IL-1 alpha, and IL-6 found in the supernatants of Listeria-infected P388D1 macrophages generally correlated well with the induction of the respective mRNAs, but it became obvious that cytokine activity is also regulated through posttranscriptional mechanisms. In vitro induction of the cytokines IL-1 alpha, IL-1 beta, IL-6, and TNF-alpha was also observed by infection of bone-marrow-derived macrophages with L. monocytogenes.