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Generation of tumoricidal cytotoxic T lymphocytes from healthy donors after in vitro stimulation with a replication‐incompetent vaccinia virus encoding MART‐1/Melan‐A 27‐35 epitope
Author(s) -
Zajac Paul,
Oertli Daniel,
Spagnoli Giulio C.,
Noppen Christoph,
Schaefer Christoph,
Heberer Michael,
Marti Walter R.
Publication year - 1997
Publication title -
international journal of cancer
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.475
H-Index - 234
eISSN - 1097-0215
pISSN - 0020-7136
DOI - 10.1002/(sici)1097-0215(19970502)71:3<491::aid-ijc30>3.0.co;2-g
Subject(s) - ctl* , cytotoxic t cell , virology , immunogenicity , epitope , biology , vaccinia , antigen , elispot , immunology , cd8 , recombinant dna , in vitro , gene , biochemistry
Active specific immunotherapy targeting tumor‐associated antigens (TAA) requires reagents of high immunogenicity and safety. To address this issue, we constructed a recombinant vaccinia virus carrying a minigene insert encoding the HLA‐A2.1‐restricted MART‐1/Melan‐A 27‐35 melanoma TAA (rVV‐M). To facilitate the entry of the antigenic epitope into the endoplasmic reticulum, a sequence coding for adenovirus E3/19K leader peptide was added. This rVV‐M was made replication‐incompetent by treatment with psoralen and UV light. Infection with rVV‐M rendered HLA‐A2.1 EBV‐transformed lymphoblastoid cells sensitive to the cytotoxic effects of HLA‐class‐I‐restricted, MART‐1/Melan‐A 27‐35 ‐specific cytotoxic T lymphocytes (CTL). The capacity of rVV‐M to generate HLA‐A2.1‐restricted MART‐1/Melan A‐specific CTL was demonstrated from tumor‐infiltrating‐lymphocyte (TIL) cultures and from healthy donors' peripheral‐blood mononuclear cells (PBMC). MART‐1/Melan‐A 27‐35 ‐specific CTL were generated from TIL after 2 weekly stimulation courses. Infection with rVV‐M elicited a higher CTL response than addition of exogenous peptide, whereas, when a similar protocol was used to stimulate PBMC of healthy donors, significant and specific cytotoxic activity could be observed only upon rVV‐M infection but not upon exogenous peptide addition. All CTL generated upon rVV‐M stimulation were also able to efficiently kill melanoma cell lines expressing both MART‐1/Melan‐A and HLA‐A2.1. In addition, TNF‐α production could be induced in rVV‐M‐stimulated CTL upon co‐culture with COS‐7 cells transiently transfected with MART‐1/Melan‐A and HLA‐A2.1 genes. This safe and highly immunogenic reagent could be of use in TAA‐targeted clinical immunotherapy. Int. J. Cancer 71:491‐496, 1997. © 1997 Wiley‐Liss Inc.