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Definition of Natural T Cell Antigens with Mimicry Epitopes Obtained from Dedicated Synthetic Peptide Libraries¹

Hoebert S. Hiemstra^*, Peter A. van Veelen^*, Nanette C. Schloot^*, Annemieke Geluk^*, Krista E. van Meijgaarden^*, Sabine J. M. Willemen^*, Jack A. M. Leunissen, Willemien E. Benckhuijsen^*, Reinout Amons, René R. P. de Vries^*, Bart O. Roep^*, Tom H. M. Ottenhoff^* and Jan W. Drijfhout^2,*

^* Department of Immunohematology and Blood Bank, Leiden University Medical Center, Leiden, The Netherlands; National Center for Computer-Aided Chemistry and Bioinformatics (CAOS/CAMM), Catholic University Nijmegen, Nijmegen, The Netherlands; and Department of Medical Biochemistry, Sylvius Laboratory, Leiden University Medical Center, Leiden, The Netherlands

Progress has recently been made in the use of synthetic peptide libraries for the identification of T cell-stimulating ligands. T cell epitopes identified from synthetic libraries are mimics of natural epitopes. Here we show how the mimicry epitopes obtained from synthetic peptide libraries enable unambiguous identification of natural T cell Ags. Synthetic peptide libraries were screened with Mycobacterium tuberculosis-reactive and -autoreactive T cell clones. In two cases, database homology searches with mimicry epitopes isolated from a dedicated synthetic peptide library allowed immediate identification of the natural antigenic protein. In two other cases, an amino acid pattern that reflected the epitope requirements of the T cell was determined by substitution and omission mixture analysis. Subsequently, the natural Ag was identified from databases using this refined pattern. This approach opens new perspectives for rapid and reliable Ag definition, representing a feasible alternative to the biochemical and genetic approaches described thus far.

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