The cytotoxic T-lymphocyte response against a poorly immunogenic mammary adenocarcinoma is focused on a single immunodominant class I epitope derived from the gp70 Env product of an endogenous retrovirus

Antonio Rosato, Silvia Dalla Santa, Alessia Zoso, Sofia Giacomelli, Gabriella Milan, Beatrice Macino, Valeria Tosello, Paolo Dellabona, Pier Luigi Lollini, Carla De Giovanni, Paola Zanovello

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Abstract

The TS/A mouse mammary adenocarcinoma is a poorly immunogenic tumor widely used in preclinical models of cancer immunotherapy. CTLs have often been indicated as important in TS/A tumor destruction, but their generation in this model has been rarely studied, nor have their precise target(s) been identified. We hypothesized that the gp70 Env product of an endogenous murine leukemia virus could be a target antigen for TS/A-specific CTLs and investigated this possibility in four different TS/A cell lines engineered with the genes that encode IFN-γ, IFN-γ interleukin-4, and B7.1, respectively. All tumor cell lines expressed gp70, albeit at different levels, as demonstrated by reverse transcription-PCR analysis. Transfected tumor cells exhibited a delayed growth in vivo, and partial tumor regression. Spleen cells from mice that displayed tumor regression had high percentages of CD8+ T cells that were specifically stained with Ld tetramers loaded with gp70423-431, the antigenic epitope of gp70 protein. Mixed leukocyte-peptide and mixed leukocyte-tumor cultures, set up by stimulating splenocytes with the immunogenic peptide and with transfected TS/A tumor cells, respectively, resulted in similar large increases in tetramer-reactive CD8+ T cells and showed high lytic activity specific for gp70423-431. Finally, in a Cold Target Inhibition assay, lytic activity of a mixed leukocyte-tumor culture was inhibited in an overlapping fashion by both the TS/A line used for restimulation and 293Ld cells loaded with gp70423-431 peptide, but not by 293Ld cells pulsed with an irrelevant H-2 Ld epitope, thus demonstrating that all or most of the cytotoxic activity was directed exclusively against this antigenic epitope.

Original languageEnglish
Pages (from-to)2158-2163
Number of pages6
JournalCancer Research
Volume63
Issue number9
Publication statusPublished - May 1 2003

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Endogenous Retroviruses
Cytotoxic T-Lymphocytes
Epitopes
Adenocarcinoma
Breast
Neoplasms
Leukocytes
Peptides
T-Lymphocytes
Murine Leukemia Viruses
Tumor Cell Line
Interleukin-4
Immunotherapy
Reverse Transcription
Spleen
Antigens
Cell Line
Polymerase Chain Reaction

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

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The cytotoxic T-lymphocyte response against a poorly immunogenic mammary adenocarcinoma is focused on a single immunodominant class I epitope derived from the gp70 Env product of an endogenous retrovirus. / Rosato, Antonio; Dalla Santa, Silvia; Zoso, Alessia; Giacomelli, Sofia; Milan, Gabriella; Macino, Beatrice; Tosello, Valeria; Dellabona, Paolo; Lollini, Pier Luigi; De Giovanni, Carla; Zanovello, Paola.

In: Cancer Research, Vol. 63, No. 9, 01.05.2003, p. 2158-2163.

Research output: Contribution to journalArticle

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abstract = "The TS/A mouse mammary adenocarcinoma is a poorly immunogenic tumor widely used in preclinical models of cancer immunotherapy. CTLs have often been indicated as important in TS/A tumor destruction, but their generation in this model has been rarely studied, nor have their precise target(s) been identified. We hypothesized that the gp70 Env product of an endogenous murine leukemia virus could be a target antigen for TS/A-specific CTLs and investigated this possibility in four different TS/A cell lines engineered with the genes that encode IFN-γ, IFN-γ interleukin-4, and B7.1, respectively. All tumor cell lines expressed gp70, albeit at different levels, as demonstrated by reverse transcription-PCR analysis. Transfected tumor cells exhibited a delayed growth in vivo, and partial tumor regression. Spleen cells from mice that displayed tumor regression had high percentages of CD8+ T cells that were specifically stained with Ld tetramers loaded with gp70423-431, the antigenic epitope of gp70 protein. Mixed leukocyte-peptide and mixed leukocyte-tumor cultures, set up by stimulating splenocytes with the immunogenic peptide and with transfected TS/A tumor cells, respectively, resulted in similar large increases in tetramer-reactive CD8+ T cells and showed high lytic activity specific for gp70423-431. Finally, in a Cold Target Inhibition assay, lytic activity of a mixed leukocyte-tumor culture was inhibited in an overlapping fashion by both the TS/A line used for restimulation and 293Ld cells loaded with gp70423-431 peptide, but not by 293Ld cells pulsed with an irrelevant H-2 Ld epitope, thus demonstrating that all or most of the cytotoxic activity was directed exclusively against this antigenic epitope.",
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AU - Zoso, Alessia

AU - Giacomelli, Sofia

AU - Milan, Gabriella

AU - Macino, Beatrice

AU - Tosello, Valeria

AU - Dellabona, Paolo

AU - Lollini, Pier Luigi

AU - De Giovanni, Carla

AU - Zanovello, Paola

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