Abstract
The transcription factor brachyury is a major driver of epithelial to mesenchymal transition in human carcinoma cells. It is overexpressed in several human tumor types versus normal adult tissues, except for testes and thyroid. Overexpression is associated with drug resistance and poor prognosis. Previous studies identified a brachyury HLA-A2 cytotoxic T-lymphocyte epitope. The studies reported here describe an enhancer epitope of brachyury. Compared to the native epitope, the agonist epitope: (a) has enhanced binding to MHC class I, (b) increased the IFN-γ production from brachyury-specific T cells, (c) generated brachyury-specific T cells with greater levels of perforin and increased proliferation, (d) generated T cells more proficient at lysing human carcinoma cells endogenously expressing the native epitope, and (e) achieved greater brachyury-specific T-cell responses in vivo in HLA-A2 transgenic mice. These studies also report the generation of a heat-killed recombinant Saccharomyces cerevisiae (yeast) vector expressing the full-length brachyury gene encoding the agonist epitope. Compared to yeast-brachyury (native) devoid of the agonist epitope, the yeast-brachyury (agonist) enhanced the activation of brachyury-specific T cells, which efficiently lysed human carcinoma cells. In addition to providing the rationale for the recombinant yeast-brachyury (agonist) as a potential vaccine in cancer therapy, these studies also provide the rationale for the use of the agonist in (a) dendritic cell (DC) vaccines, (b) adjuvant or liposomal vaccines, (c) recombinant viral and/or bacterial vaccines, (d) protein/polypeptide vaccines, (e) activation of T cells ex vivo in adoptive therapy protocols, and (f) generation of genetically engineered targeted T cells.
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Abbreviations
- ATCC:
-
American Type Culture Collection
- BFA:
-
Brefeldin A
- BLAST:
-
Basic local alignment search tool
- CTL:
-
Cytotoxic T lymphocyte
- DCs:
-
Dendritic cells
- EMT:
-
Epithelial to mesenchymal transition
- IHC:
-
Immunohistochemistry
- MFI:
-
Mean fluorescence intensity
- MHC:
-
Major histocompatibility complex
- PBMCs:
-
Peripheral blood mononuclear cells
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Acknowledgments
Grant support was provided by the Intramural Research Program of the Center for Cancer Research, National Cancer Institute, National Institutes of Health, and a Cooperative Research and Development Agreement between the National Cancer Institute and GlobeImmune, Inc. The authors thank Diane J. Poole, Garland Davis and Curtis Randolph for technical assistance and Debra Weingarten for editorial assistance in the preparation of this manuscript.
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The authors declare that they have no conflicts of interest.
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Jo A. Tucker and Caroline Jochems have contributed equally to this paper as primary authors. Jeffrey Schlom and Kwong-Yok Tsang have contributed equally to this paper as senior authors.
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Tucker, J.A., Jochems, C., Boyerinas, B. et al. Identification and characterization of a cytotoxic T-lymphocyte agonist epitope of brachyury, a transcription factor involved in epithelial to mesenchymal transition and metastasis. Cancer Immunol Immunother 63, 1307–1317 (2014). https://doi.org/10.1007/s00262-014-1603-2
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DOI: https://doi.org/10.1007/s00262-014-1603-2