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. Author manuscript; available in PMC: 2019 Sep 1.
Published in final edited form as: J Immunol. 2018 Jul 16;201(5):1536–1548. doi: 10.4049/jimmunol.1701240

Figure 1. Generation of retrogenic graft-specific CD8+ T cells that constitutively express 2B4.

Figure 1

A, Frequencies of GFP+ (pMY control vector) or GFP+ 2B4+ (for 2B4 vector) CD45.2+ Thy1.1+ OT-I BM cells at d2. B, BM cells were adoptively transferred into irradiated CD45.1+ Thy1.2+ animals and were detectable at 8–10 weeks. C, GFP+ CD8+ Thy1.1+ (pMY) and GFP+ 2B4+ CD3+ Thy1.1+ (2B4rg) OT-I T cells sorted and D, adoptively transferred (106/recipient) into naïve B6 hosts that received two OVA-expressing skin grafts 48 hours later. Data shown in A–D are representative of 11-12 mice/group from 3 independent experiments.

E, 2B4 expression is maintained on the retrovirally transduced retrogenic cells following transplantation. F, 106 WT CD8+ Thy1.1+ or GFP+ 2B4+ CD3+ Thy1.1+ (2B4rg) OT-I T cells were adoptively transferred naïve B6 hosts 48 hours prior to receiving two OVA-expressing skin grafts. Animals received 250 μg of CTLA4Ig on post-operative Day 0, 2, 4, and 6. Graft survival was measured, and rejection was scored based on a loss of +90% viable tissue. Data shown in Figure 1 A–E are representative of 11-12 mice/group from 3 independent experiments. ***p<0.0001. Data shown in Figure 1F are representative of 12-18 mice/group from 2 independent experiments.