Fig. 1.

TCR-transgenic humanized model for the study of human T-cell selection. a. CD34⁺ cells are MACS-sorted from HLA-DQ8⁺ or HLA-DQ8^- human fetal liver tissue, then lentivirally transduced with an insulin B:9-23-reactive TCR containing a GFP reporter or with vector containing a GFP reporter alone. HLA-DQ8⁺ human fetal thymus tissue is isolated and T-cell-depleted by freeze-thawing. b. Thymectomized 8–10 week old NSG mice receive TBI on day −1. Next, the mice are injected intratibially with transduced CD34⁺ cells from (A). Mice are co-transplanted with a fragment of fetal thymus from (A) under the kidney capsule. Mice are injected with anti-CD2 antibody on days 1 and 7 to deplete any T cells escaping from the thymus graft. The human immune system reconstitution in peripheral blood is then monitored every two weeks until sacrifice. c, d. Representative FCM analysis of lentiviral transduction efficiency of FLCs measured by GFP expression. e-j. Peripheral blood reconstitution of human immune cell subsets in mice transplanted with Clone 5 or GFP-only virus is shown over time post-transplantation. e. Proportions of huCD45⁺ cells f. % GFP⁺ of huCD45⁺, g. Proportions of human B cells; h. Proportions of human T cells, i. Proportions of human CD4⁺ T cells, and j. Proportions of human CD8⁺ T cells. Data are a compilation of n ​= ​4 Clone 5 and n ​= ​3 GFP-only mice transplanted with CD34⁺ and thymus tissue from a single HLA-DQ8⁺ fetal donor and n ​= ​4 Clone 5, n ​= ​4 GFP-only mice transplanted with HLA-DQ8^- CD34⁺ cells and HLA-DQ8⁺ thymus.