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. 2013 Sep 4;4:270. doi: 10.3389/fimmu.2013.00270

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Copyright © 2013 Schaft, Coccoris, Drexhage, Knoop, de Vries, Adema and Debets.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Differential T cell responsiveness toward gp100 APLs is related to TCR’s ability to bind gp100/HLA-A2 complexes. TCR-engineered T cells were stained with titrated amounts of PE-conjugated peptide-MHC multimers (range: 400–12,5 nM peptide-MHC complexes) and analyzed via flow cytometry. The gp100-HLA-A2 multimers comprise either gp100 wt peptide or APLs A3 and S4S6. The BMLF-1-HLA-A2 multimer served as a negative control. Binding of peptide-MHC multimers is analyzed as described in Section “Flow Cytometry and Cell-Sorting” and indicated in percentage. Percentages correspond to marker-positive T cells. Results of one (out of two) representative experiment are shown based on the mean value of triplicate data points.