Fig. 6: Sm-TCR lentiviral transduction produces high-affinity, antigen-specific responses.
From: Smith-specific regulatory T cells halt the progression of lupus nephritis
a lentiviral construct design showing from 5’ to 3’ the Sm-TCR beta chain, P2A ribosomal skipping sequence, Sm-TCR alpha chain, T2A ribosomal skipping sequence, and eGFP reporter under the control of a 5’ EF1-alpha promoter and 3’ WPRE. b Saturation binding curves of the top three SmB/B’58-72-specific TCRs (TCR1 to TCR3) showing the apparent affinity KD (nM) and the maximum specific binding in units of Dextramer PE mean fluorescence intensity (MFI) (Bmax). TCR1-3 show DR15/ SmB/B’58-72 dextramer binding and negative control (Ctrl, dotted gray) shows DR15/ CLIP103-117 dextramer that is not expected to bind. c–h Image flow cytometry of the interaction of c TCR1, e TCR2, g TCR3 on J76 Jurkats incubated for 2 h with HLA-DR15 B-LCLs in the presence (above) and absence (below) of SmB/B’58-72. Arrows point to the mature immune synapse (IS). Quantification of the CD3 and F-actin MFI at the IS of (d), TCR1 (n = 66 IS cells with SmB/B’58-72, n = 86 without SmB/B’58-72) CD3 P = < 0.000001, F-actin P = 0.000409, f TCR2 (n = 47 IS cells with SmB/B’58-72, n = 57 without SmB/B’58-72) CD3 P = 0.014402, F-actin P = 0.000998, h TCR3 (n = 111 IS with SmB/B’58-72, n = 83 without SmB/B’58-72) CD3 P = 0.000720, F-actin P = 0.000004. J76 Jurkats and HLA-DR15 B-LCLs in the presence (red) or absence (brown) of SmB/B’58-72. Data are presented as mean with SD. *P < 0.05, **P < 0.01, ***P < 0.001 by Mann–Whitney test. Source data are provided as a Source Data file.
