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Single and Dual Amino Acid Substitutions in TCR CDRs Can Enhance Antigen-Specific T Cell Functions

Paul F. Robbins^1,*, Yong F. Li^*, Mona El-Gamil^*, Yangbing Zhao^*, Jennifer A. Wargo^*, Zhili Zheng^*, Hui Xu^*, Richard A. Morgan^*, Steven A. Feldman^*, Laura A. Johnson^*, Alan D. Bennett, Steven M. Dunn, Tara M. Mahon, Bent K. Jakobsen and Steven A. Rosenberg^*

^* Surgery Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892; and MediGene Limited, Abingdon, Oxfordshire, United Kingdom

Single and dual amino acid substitution variants were generated in the TCR CDRs of three TCRs that recognize tumor-associated Ags. Substitutions that enhance the reactivity of TCR gene-modified T cells to the cognate Ag complex were identified using a rapid RNA-based transfection system. The screening of a panel of variants of the 1G4 TCR, that recognizes a peptide corresponding to amino acid residues 157–165 of the human cancer testis Ag NY-ESO-1 (SLLMWITQC) in the context of the HLA-A*02 class I allele, resulted in the identification of single and dual CDR3 and CDR2β amino acid substitutions that dramatically enhanced the specific recognition of NY-ESO-1⁺/HLA-A*02⁺ tumor cell lines by TCR gene-modified CD4⁺ T cells. Within this group of improved TCRs, a dual substitution in the 1G4 TCR CDR3 chain was identified that enhanced Ag-specific reactivity in gene-modified CD4⁺ and CD8⁺ T cells. Separate experiments on two distinct TCRs that recognize the MART-1 27–35 (AAGIGILTV) peptide/HLA-A*02 Ag complex characterized single amino acid substitutions in both TCRs that enhanced CD4⁺ T cell Ag-specific reactivity. These results indicate that simple TCR substitution variants that enhance T cell function can be identified by rapid transfection and assay techniques, providing the means for generating potent Ag complex-specific TCR genes for use in the study of T cell interactions and in T cell adoptive immunotherapy.

The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

¹ Address correspondence and reprint requests to Dr. Paul F. Robbins, Surgery Branch, National Cancer Institute, National Institutes of Health, 10 Center Drive, CRC 3-5744, Bethesda, MD 20892-1201. E-mail address: Paul_Robbins{at}nih.gov

² Abbreviations used in this paper: AAS, amino acid substitution; WT, wild type.

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The JI 2008 180: 5759-5760. [Full Text]