Figure 2.
Figure 2. CD8+ Tregs and Tcons have similar T-cell receptor repertoire diversity. Lethally irradiated (900 cGy) Balb/c mice were transplanted with B6 Rag-1 BM (5 × 106) and flow-sorted CD4+ Foxp3− (0.4 × 106) and CD8+ Foxp3− (0.2 × 106) T cells from B6 Foxp3EGFP animals. Mice were euthanized on day 7 posttransplantation, and donor-derived CD4+ Foxp3+, CD8+ Foxp3+, CD4+ Foxp3−, and CD8+ Foxp3− T cells were sorted from the spleens and livers of recipients. (A) Shannon entropy of CD4+ Foxp3+ (n = 4), CD4+ Foxp3− (n = 6), CD8+ Foxp3+ (n = 6), and CD8+ Foxp3− (n = 6) T-cell populations. Error bars represent standard error of the mean. (B) Morsita Horn similarity index of CD4+ Tregs and Tcons (n = 4) and CD8+ Treg and Tcons (n = 6). Colored circles denote individual mice. (C) Heat map showing Morsita Horn diversity overlap of TCRβ clonotype expression in CD4+ and CD8+ Treg and Tcon populations from individual mice. (D) Lethally irradiated (900 cGy) Balb.B mice were transplanted with B6 Foxp3EGFP BM (5 × 106) and spleen cells (adjusted to yield a dose of 20 × 106 αβ T cells). Animals were euthanized on day 7 posttransplantation, and cells were harvested from the liver and spleen. Representative contour plot depicting CD8+ H60+ Foxp3+ and CD8+ H60+ Foxp3− T cells. (E) Shannon entropy of CD8+ H60+ Treg (n = 2) and Tcon (n = 2) populations. Error bars represent standard error of the mean. (F) Heat map showing Morsita Horn diversity overlap of TCRβ clonotype expression in CD8+ H60+ Foxp3+ and Foxp3− T cells from individual mice. *P < .05; **P < .01.

CD8+Tregs and Tcons have similar T-cell receptor repertoire diversity. Lethally irradiated (900 cGy) Balb/c mice were transplanted with B6 Rag-1 BM (5 × 106) and flow-sorted CD4+ Foxp3 (0.4 × 106) and CD8+ Foxp3 (0.2 × 106) T cells from B6 Foxp3EGFP animals. Mice were euthanized on day 7 posttransplantation, and donor-derived CD4+ Foxp3+, CD8+ Foxp3+, CD4+ Foxp3, and CD8+ Foxp3 T cells were sorted from the spleens and livers of recipients. (A) Shannon entropy of CD4+ Foxp3+ (n = 4), CD4+ Foxp3 (n = 6), CD8+ Foxp3+ (n = 6), and CD8+ Foxp3 (n = 6) T-cell populations. Error bars represent standard error of the mean. (B) Morsita Horn similarity index of CD4+ Tregs and Tcons (n = 4) and CD8+ Treg and Tcons (n = 6). Colored circles denote individual mice. (C) Heat map showing Morsita Horn diversity overlap of TCRβ clonotype expression in CD4+ and CD8+ Treg and Tcon populations from individual mice. (D) Lethally irradiated (900 cGy) Balb.B mice were transplanted with B6 Foxp3EGFP BM (5 × 106) and spleen cells (adjusted to yield a dose of 20 × 106 αβ T cells). Animals were euthanized on day 7 posttransplantation, and cells were harvested from the liver and spleen. Representative contour plot depicting CD8+ H60+ Foxp3+ and CD8+ H60+ Foxp3 T cells. (E) Shannon entropy of CD8+ H60+ Treg (n = 2) and Tcon (n = 2) populations. Error bars represent standard error of the mean. (F) Heat map showing Morsita Horn diversity overlap of TCRβ clonotype expression in CD8+ H60+ Foxp3+ and Foxp3 T cells from individual mice. *P < .05; **P < .01.

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