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A Long-Term Memory Obtained by Genetic Immunization Results in Full Protection from a Mammary Adenocarcinoma Expressing an EBV Gene¹

Jehad Charo^2,*,, Anne-Marie T. Ciupitu^*,, Alain Le Chevalier de Préville^*, Pankaj Trivedi^3,, George Klein, Jorma Hinkula and Rolf Kiessling^*,

^* Cancer Center Karolinska (CCK), Karolinsk Hospital, Stockholm, Sweden; Microbiolgy and Tumorbiology Center (MTC), Karolinska Institute, Stockholm, Sweden; and Swedish Institute for Infectious Disease Control, Stockholm, Sweden

We have tested the capability of a plasmid DNA (pDNA) expressing the EBV nuclear Ag-4 (EBNA-4) to evoke a T cell response-associated protective immune response against a tumor expressing this gene. We have found that ACA mice immunized with EBNA-4-expressing plasmid were partially protected against syngeneic mammary carcinoma line (S6C) expressing EBNA-4 (S6C-E4). This protection was enhanced by coimmunizing mice with EBNA-4- and GM-CSF-expressing plasmids, and a full protection was achieved by coimmunizing mice with EBNA-4- and IFN--expressing plasmids. Furthermore, mice that have rejected the EBNA-4-positive tumor were also resistant against a subsequent challenge with the original nontransfected tumor line. We then checked for the ability of pDNA immunization to provide a protective long-term memory response. We indeed found that even after 3 mo from the last immunization, full protection was obtained by this method, as compared with full tumor outgrowth in the control-immunized group. These findings support the concept that a nonviral, pDNA-based vaccination strategy is useful to fully protect from the outgrowth of tumors expressing this EBV gene.

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