Immunodominant West Nile Virus T Cell Epitopes Are Fewer in Number and Fashionably Late

Kaabinejadian S, et al., 196: 4263-4273, J Immunol, 2016

Described herein is the generation of a panel of TCR mimic (TCRm) mAbs to characterize three HLA-A2/ West Nile virus (WNV) T cell epitopes. The binding interactions of each TCRm mAb (RL14C, RL15A, and RL26A) against all three Ags (SLF9/HLA-A2, SVG9/HLA-A2, and YTM9/HLA-A2) were studied by Surface Plasmon Resonance (SPR) spectroscopy using a Pall ForteBio Pioneer FE instrument. Using the amine coupling chemistry, Protein G was immobilized onto a COOH2 sensor chip. Assays involved binding of the RL14C, RL15A, and RL26A TCRm mAbs to protein G and one-step gradient injection of Ags to loaded Abs. The Protein G surface was regenerated with 20 mM K3PO4 (pH 12). The association rate constants (ka) and the dissociation rate constants (kd) were measured. The binding affinity (KD) of each TCRm mAb to cognate peptide/HLA-A2 complexes was determined. Overall findings of this study have enabled the authors to identify differences in the timing and levels of epitope availability in WNV infected cells.

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