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Department of Immunology, Institute for Cell Biology, University of Tübingen, Tübingen, Germany; and
Laboratory of Cellular Physiology and Immunology, The Rockefeller University, New York, NY 10021
PBL from HLA-A2- or HLA-A3- donors were stimulated with synthetic peptide libraries fitting HLA-A2 or HLA-A3 motifs and presented on HLA-A2- or HLA-A3-expressing TAP- cells. Peptide library-specific allorestricted CTL were found to constitute up to half the alloreactive CTL response and occurred at twofold lower frequency than autologous peptide library-specific CTL. This indicates that positive selection by one particular MHC class I molecule is not absolutely essential for the generation of CTL restricted to the same molecule. However, positive selection increases their frequency. The CTL obtained differed greatly both with respect to peptide dependency and peptide specificity. Determination of the peptide avidity for one representative CTL clone, 10F4, proved that the method described here allows the stimulation of high avidity cytotoxic T cells. This approach involving in vitro stimulation of T cells restricted toward a MHC molecule that was not present during their negative selection might therefore offer the possibility of isolating CTL against self and foreign peptides with varying avidities. Such T cells might indeed be useful for tumor immunotherapy.
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