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Her-2/neu altered peptide ligand–induced CTL responses: implications for peptides with increased HLA affinity and T-cell-receptor interaction

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Abstract

In this study, we developed two Her-2/neu-derived E75 altered peptide ligands (APLs) that demonstrate increased affinities for the HLA-A*0201 allele compared with wild-type E75 peptide. The APLs contain amino acids from E75(369–377), an immunodominant Her-2/neu-derived peptide, and preferred primary and auxiliary HLA-A*0201 molecule anchor residues previously identified from combinatorial peptide library screening with the recombinant molecule. CTL lines were generated against wild-type E75 peptide (KIFGSLAFL) and APLs by multiple rounds of peptide stimulation of peripheral blood mononuclear cells (PBMCs) from HLA-A2+ antigen normal individuals. CTL lines raised on wild-type E75 peptide cross-reacted with APLs and similarly, CTL lines raised on APLs cross-reacted with wild-type E75 peptide, as measured by IFN-γ ELISpot and target cell lysis assays. One of five individuals demonstrated specificity for APL 2 (FLFGSLAFL), whereas APL 5 (FLFESLAFL)-specific responses were observed from all five individuals tested. Molecular models of the E75, APL 2, and APL 5/HLA-A2 complexes indicated that the substitution of glycine with glutamic acid at position four of APL 5 resulted in the presentation of a large, negatively charged side chain that interacts with the outer edge of the HLA-A2 antigen alpha helix and is freely available to interact with cognate T-cell receptors. The results of this study further substantiate the concept that rational design of T-cell epitopes may lead to stronger peptide immunogens than natural, wild-type peptides.

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Acknowledgements

We are grateful to Dr Sue Roberts for her contributions to the molecular modeling of HLA-A2/peptide complexes. This study was supported by a National Cancer Institute grant, RO1 CA94852.

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Authors and Affiliations

  1. Department of Microbiology and Immunology, Arizona Cancer Center, University of Arizona, 1515 North Campbell Avenue, Tucson, AZ 85724, USA

    Sara O. Dionne, Cheryl E. Myers & Douglas F. Lake

  2. Department of Biochemistry, University of Arizona, Tucson, AZ 85721, USA

    Margaret H. Smith

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  1. Sara O. Dionne
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  2. Cheryl E. Myers
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  3. Margaret H. Smith
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  4. Douglas F. Lake
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Correspondence to Douglas F. Lake.

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Dionne, S.O., Myers, C.E., Smith, M.H. et al. Her-2/neu altered peptide ligand–induced CTL responses: implications for peptides with increased HLA affinity and T-cell-receptor interaction. Cancer Immunol Immunother 53, 307–314 (2004). https://doi.org/10.1007/s00262-003-0439-y

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