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Induction of Antigen-Specific CTL Responses Using Antigens Conjugated to Short Peptide Vectors

Fiona H. Day^1,*, Yu Zhang^*, Philippe Clair, Kenneth H. Grabstein^*, Martine Mazel, Anthony R. Rees, Michel Kaczorek and Jamal Temsamani

^* Corixa Corporation, Seattle, WA 98104; and Synt:em, Parc Scientifique Georges Besse, Nimes, France

Linear peptides (SynB vectors) with specific sequence motifs have been identified that are capable of enhancing the transport of a wide range of molecules into cells. These peptide vectors have been used to deliver exogenous peptides and protein Ags across the cell membrane and into the cytoplasm of cells. Specifically, in vitro analysis indicated that these SynB peptides enhanced the uptake of two 9-mer peptide Ags, NP_147–155 and Mtb_250–258 (T cell epitopes of influenza nucleoprotein and Mycobacterium tuberculosis, respectively) and the M. tuberculosis Ag Mtb8.4 protein, into K562 cells when covalently linked to the respective Ags. Furthermore, selected SynB vectors, when conjugated to these same Ags and used as immunogens, resulted in considerably enhanced Ag-specific CTL responses. Several SynB vectors were tested and resulted in varying levels of cellular uptake. The efficiency of uptake correlated with the ability of the SynB construct to deliver each epitope in vivo and induce specific CTL responses in mice. These data suggest that peptide vectors, such as SynB that transport target Ags across the cell membrane in a highly efficient manner, have significant potential for vaccine delivery.