Figure 2 In vitro refolded human leukocyte antigen (HLA)–A2:peptide complexes activate TZR 2D1 In vitro refolded human leukocyte antigen (HLA)–A2:peptide.

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Figure 2 In vitro refolded human leukocyte antigen (HLA)–A2:peptide complexes activate TZR 2D1 In vitro refolded human leukocyte antigen (HLA)–A2:peptide complexes activate TZR 2D1 (A) Sodium dodecyl sulfate polyacrylamide gel electrophoresis of HLA-A*02:01 (HLA-A2) refolded from recombinant heavy chain and β2m in the presence of either the octameric glycerolphosphatidylcholine phosphodiesterase 1 (GPCPD1) (15-22) or the nonameric M-GPCPD1(15-22) under reducing and nonreducing conditions. Lanes 1 and 6, protein standard. Molecular masses are given in kDa at the left; lane 2, HLA-A2, β2m, and M-GPCPD1(15-22) under reducing conditions; lane 3, HLA-A2, β2m, and GPCPD1(15-22) under reducing conditions; lane 4, HLA-A2, β2m, and M-GPCPD1(15-22) under nonreducing conditions; lane 5, HLA-A2, β2m, and GPCPD1(15-22) under nonreducing conditions. (B) Activation of 58α−β− T hybridoma cells expressing TCR 2D1, human CD8αβ molecules, and sGFP under the control of nuclear factor of activated T cells (58-2D1-CD8-sGFP) cells by in vitro refolded trimeric HLA-A2:β2m:peptide complexes as measured by fluorescence microscopy. Left panel: 58-2D1-CD8-sGFP cells were incubated with refolded and biotinylated HLA-A2, β2m, and GPCPD1(15-22). Right panel: As left panel, but with M-GPCPD1(15-22). Scale bars: 100 μm. (C) IL-2 concentration measured in the supernatant from the experiments shown in (B). Error bars represent SD of the mean. Geraldine Rühl et al. Neurol Neuroimmunol Neuroinflamm 2016;3:e241 © 2016 American Academy of Neurology