Anthrax toxin-mediated delivery of a cytotoxic T-cell epitope in vivo.
The protective antigen (PA) component of anthrax toxin mediates entry of the toxin's lethal factor (LF) and edema factor into the cytosolic compartment of mammalian cells. The amino-terminal domain of LF (LFn; 255 amino acids) binds LF to PA, and when fused to heterologous proteins, the LFn domain delivers such proteins to the cytoplasm in the presence of PA. In the current study, we fused a 9-amino acid cytotoxic T-lymphocyte (CTL) epitope (LLO91-99) from an intracellular pathogen, Listeria monocytogenes, to LFn and measured the ability of the resulting LFn-LLO91-99 fusion protein to stimulate a CTL response against the epitope in BALB/c mice. As little as 300 fmol of fusion could stimulate a response. The stimulation was PA-dependent and occurred with the peptide fused to either the amino terminus or the carboxyl terminus of LFn. Upon challenge with L. monocytogenes, mice previously injected with LFn-LLO91-99 and PA showed a reduction of colony-forming units in spleen and liver, relative to nonimmunized control mice. These results indicate that anthrax toxin may be useful as a CTL-peptide delivery system for research and medical applications.
Ballard
J D
JD
Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, MA 02115, USA.
Collier
R J
RJ
Starnbach
M N
MN
D000818:Animals; D000881:Anthrax; D000942:Antigens, Bacterial; D001408:Bacillus anthracis; D001427:Bacterial Toxins / Q000378:metabolism*; D018984:Epitopes, T-Lymphocyte / Q000378:metabolism*; D008089:Listeria monocytogenes; D051379:Mice; D008807:Mice, Inbred BALB C; D008954:Models, Biological; D013602:T-Lymphocytes, Cytotoxic / Q000378:metabolism*
D000942:Antigens, Bacterial; D001427:Bacterial Toxins; D018984:Epitopes, T-Lymphocyte; C030325:anthrax toxin