title: Intratumoral electroporation of IL-12 cDNA eradicates established melanomas by Trp2(180-188)-specific CD8+ CTLs in a perforin/granzyme-mediated and IFN-γ-dependent manner: application of Trp2(180-188) peptides.
abstract: Intratumoral electroporation (IT-EP) with IL-12 cDNA (IT-EP/IL12) can lead to the eradication of established B16 melanoma tumors in mice. Here, we explore the immunological mechanism of the antitumor effects generated by this therapy. The results show that IT-EP/IL12 applied only once resulted in eradication in 70% animals with large established B16 tumors. Tumor eradication required the participation of CD8+ T cells, but not CD4+ T cells and NK cells. IT-EP/IL12 induced antigen-specific CD8+ T cell responses against the immunodominant Trp2(180-188) epitope and generated a systemic response, resulting in significant therapeutic effects against distal, untreated tumors. The therapeutic effect of IT-EP/IL12 was absent in perforin-deficient mice, indicating that tumor elimination occurred through conventional perforin/granzyme lysis by CTLs. Moreover, this therapy induced some degree of immunological memory that protected approximately one-third of the cured mice against a subsequent tumor challenge. Moreover, antitumor efficacy and long-term protection against B16 were significantly improved by concurrent Trp2 peptide immunization through more induction of Ag-specific CTL responses and more attraction of IFN-γ-expressing CD8+ T cells into tumor sites. The antitumor effect of IT-EP/IL12 required the participation of IFN-γ, which was shown to induce MHC class I expression on B16 cells and increase the lytic activity of the CD8+ CTL generated by IT-EP/IL12. The results from these animal studies may help in the development of IT-EP/IL12 for cancer patients.
authors.affiliation: Department of Microbiology, School of Medicine, Kangwon National University, Chuncheon, Gangwon-do 200-701, Korea. jsin1964@hanmail.net
authors.forename: Jeong-Im
authors.initials: JI
authors.lastname: Sin
authors.forename: Jae-Bok
authors.initials: JB
authors.lastname: Park
authors.forename: In Hee
authors.initials: IH
authors.lastname: Lee
authors.forename: Daehan
authors.initials: D
authors.lastname: Park
authors.forename: Youn Seok
authors.initials: YS
authors.lastname: Choi
authors.forename: Jongseon
authors.initials: J
authors.lastname: Choe
authors.forename: Esteban
authors.initials: E
authors.lastname: Celis
mesh_terms: D000818:Animals; D018076:DNA, Complementary / Q000235:genetics; D004195:Disease Models, Animal; D053672:Electrochemotherapy / Q000379:methods*; D005260:Female; D053804:Granzymes / Q000276:immunology*; D007156:Immunologic Memory; D007371:Interferon-gamma / Q000276:immunology*; D018664:Interleukin-12 / Q000235:genetics* / Q000276:immunology; D019746:Intramolecular Oxidoreductases / Q000276:immunology*; D008546:Melanoma, Experimental / Q000628:therapy*; D051379:Mice; D052899:Pore Forming Cytotoxic Proteins / Q000276:immunology*; D012878:Skin Neoplasms / Q000628:therapy*; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*
chemical_list: D018076:DNA, Complementary; D052899:Pore Forming Cytotoxic Proteins; C483886:perforin, mouse; D018664:Interleukin-12; D007371:Interferon-gamma; D053804:Granzymes; D019746:Intramolecular Oxidoreductases; C042501:dopachrome isomerase