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. 2012 Jan 10;20(3):652–660. doi: 10.1038/mt.2011.286

Figure 3.

Figure 3

m33 TCR-expressing CD8+ T cells are rapidly deleted in vivo. (a) Blood of OT-I/Thy1.1 recipient mice were analyzed 3 days after adoptive transfer of 2C or m33-transduced OT-I T cells. Leukocytes were stained with anti-Thy1.1, anti-CD8, and anti-Vβ8. Left panels show cells gated on Thy1.1– lymphocytes, numbers indicate the percentage of cells of all Thy1.1– cells. The right panels show Thy1.1–, CD8+ lymphocytes and the numbers indicate the percentage of Vβ8+ cells among Thy1.1– CD8+ cells. Similar results were obtained in three experiments, showing a mean of 21% of CD8+ T cells expressing 2C (ranging from 10 to 46%) and only 0.8% expressing m33 (ranging from 0 to 1%). (b) Analysis of liver and lung of OT-I/Thy1.1 mice that received 2C or m33-transduced OT-I T cells 6 hours earlier. Leukocytes were stained with anti-Thy1.1, anti-CD8, anti-Vβ8. Cells shown were gated on Thy1.1– CD8+ lymphocytes, the numbers in each panel indicate percentage of Vβ8+ cells among Thy1.1– CD8+ cells. Data shown are representative for three independent experiments. (c) Analysis of OT-I/Thy1.1 mice described in (b) on day 3 after transfer. The left graphs show Thy1.1– CD8+ lymphocytes; the numbers in each panel indicate the percentage of Vβ8+ cells among Thy1.1– CD8+ cells. The right graphs are additionally gated on Vβ8+ cells and numbers indicate the percentage of Annexin V-positive Thy1.1– CD8+ Vβ8+ cells. (d) Blood of OT-I/Thy1.1 recipient mice was analyzed 2 days after adoptive transfer of m33-transduced Bcl-2-, Bcl-xL-transgenic or wild-type T cells (m33 expression pretransfer was 19, 12, and 33%, respectively). Leukocytes were stained for CD8, Thy1.1, and with (SIY-Kb)2/IgG. Cells shown in the dot plots were gated on Thy1.1– cells. Numbers in each panel indicate percentage of SIY-Kb-binding T cells among Thy1.1– CD8+ T cells. (e) Blood of Rag–/–Kb–/–Db–/– and Rag–/– recipient mice was analyzed 3 days after adoptive transfer of m33- or 2C-transduced OT-I T cells. Leukocytes were stained for CD8 and Vβ8. Cells shown in the histograms were gated on CD8+ cells. Numbers in each panel indicate percentage of Vb8+ cells among CD8+ T cells.