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CTL recognition of a novel HLA-A*0201-binding peptide derived from glioblastoma multiforme tumor cells

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Abstract

Genetic instability of tumor cells can result in translation of proteins that are out of frame, resulting in expression of neopeptides. These neopeptides are not self-proteins and therefore should be immunogenic. By eluting peptides from human glioblastoma multiforme (GBM) tumor cell surfaces and subjecting them to tandem mass spectrometry, we identified a novel peptide (KLWGLTPKVTPS) corresponding to a frameshift in the 3′ beta-hydroxysteroid dehydrogenase type 7 (HSD3B7) gene. HLA-binding algorithms predicted that a 9-amino acid sequence embedded in this peptide would bind to HLA-A*0201. We confirmed this prediction using an HLA-A*0201 refolding assay followed by live cell relative affinity assays, but also showed that the 12-mer binds to HLA-A*0201. Based on the 9-mer sequence, optimized peptide ligands (OPL) were designed and tested for their affinities to HLA-A*0201 and their abilities to elicit anti-peptide and CTL capable of killing GBM in vitro. Wild-type peptides as well as OPL induced anti-peptide CTL as measured by IFN-γ ELISPOTS. These CTL also killed GBM tumor cells in chromium-51 release assays. This study reports a new CTL target in GBM and further substantiates the concept that rational design and testing of multiple peptides for the same T-cell epitope elicits a broader response among different individuals than single peptide immunization.

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Acknowledgments

We would like to thank HoJoon Lee, M.S. for the frameshift database.

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Authors and Affiliations

  1. School of Life Sciences, Arizona State University, Tempe, AZ, 85287, USA

    Cheryl E. Myers, Paul Hanavan, Kwasi Antwi & Douglas F. Lake

  2. Department of Chemistry and Biochemistry, Arizona State University, Tempe, AZ, 85287, USA

    Zachary Laughrey

  3. Department of Medicine, Arizona Cancer Center, University of Arizona, Tucson, AZ, 85724, USA

    Daruka Mahadevan, A. Jamal Nadeem & Laurence Cooke

  4. Barrow Neurological Institute of SJHMC, Phoenix, AZ, 85013, USA

    Adrienne C. Scheck

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  1. Cheryl E. Myers
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  9. Douglas F. Lake
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Correspondence to Douglas F. Lake.

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Myers, C.E., Hanavan, P., Antwi, K. et al. CTL recognition of a novel HLA-A*0201-binding peptide derived from glioblastoma multiforme tumor cells. Cancer Immunol Immunother 60, 1319–1332 (2011). https://doi.org/10.1007/s00262-011-1032-4

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  • DOI: https://doi.org/10.1007/s00262-011-1032-4

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