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Immunization with gp96 from Listeria monocytogenes-Infected Mice Is Due to N-Formylated Listerial Peptides¹

Anne-Marit Sponaas^*, Ulrich Zuegel, Stephan Weber^*, Robert Hurwitz^*, Ralf Winter, Stephanie Lamer, Peter R. Jungblut and Stefan H. E. Kaufmann^*

^* Department of Immunology, Central Support Unit Biochemistry, Max-Planck Institute for Infection Biology, Berlin, Germany; and Department of Experimental Dermatology, Schering AG, Berlin, Germany

N-Formylated (N-f-met) peptides derived from proteins of the intracellular bacterium Listeria monocytogenes generate a protective, H2-M3-restricted CD8 T cell response in C57BL/6 mice. N-f-met peptide-specific CTL were generated in vitro when mice previously immunized with gp96 isolated from donor mice infected with L. monocytogenes were stimulated with these peptides. No significant peptide-specific CTL activity was observed in mice immunized with gp96 from uninfected animals. Masses corresponding to one N-f-met peptide were found by matrix-assisted laser desorption/ionization-mass spectrometry on gp96 isolated from C57BL/6 mice infected with L. monocytogenes, but not on gp96 from noninfected mice. Therefore, bacterial N-f-met peptides from intracellular bacteria can bind to gp96 in the infected host, and gp96 loaded with these peptides can generate N-f-met-peptide-specific CTL. We assume a unique role of gp96 in Ag processing through the H2-M3 pathway.

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