Fig. 1: Adoptively transferred ex vivo cultured Tregs have a limited lifespan in mice. | Nature Communications

Fig. 1: Adoptively transferred ex vivo cultured Tregs have a limited lifespan in mice.

From: Expression of an interleukin-2 partial agonist enhances regulatory T cell persistence and efficacy in mouse autoimmune models

Fig. 1

A Schematic experimental design. Created in BioRender49. Tregs were isolated from the spleen and lymph nodes of male or female B6 Thy1.1 mice at six to twenty weeks of age. On day 5 of culture, the cells were transduced with the control vector MSCV-Thy1.2 for the expression of the reporter gene mouse Thy1.2 alone in the transduced cells. Two days post-transduction, 9 × 105 unsorted transduced control Tregs (Ctrl Tregs) per mouse were injected into B6 Thy1.2 mice aged six to twenty weeks. Recipient mice were sex-matched with the donor cells. Survival of the injected cells was tracked in the blood or organs of the mice, as indicated. Contour plots representing transduction efficacy and Treg purity on the day of cell injection are shown on the left. B Gating strategy for tracking Thy1.1 donor cells within the CD3+CD4+ population of Thy1.2 recipient mice. Transduced and non-transduced donor cells are Thy1.1+Thy1.2+ and Thy1.1+Thy1.2-, respectively; endogenous CD4 cells are Thy1.2+Thy1.1-. A zebra plot from the blood of a mouse receiving Ctrl Tregs 14 days post-injection is shown. Schematic created in BioRender49. C Dynamics of Thy1.1 donor Ctrl Tregs in the blood. Data were extracted from one representative out of four experiments with a total of n = 8. D Dynamics of Thy1.1 injected Ctrl Tregs in the spleen, lungs, and liver. E, F Dynamics of Ctrl Tregs (E) and endogenous Tregs (F) in the blood of recipient mice treated with an AAV-IL2 (orange, n = 3) or not (green, n = 8), from four independent experiments. Statistically significant differences were found between Ctrl Tregs injected into mice treated with AAV-IL2 and untreated mice on day 20 (p = 0.0357) and day 55 (p = 0.0250), and between endogenous Tregs in AAV-IL2-treated and untreated mice on days 14 (p = 0.061), 20 (p = 0.0119), 27 (p = 0.0121), 43 (p = 0.0167), 55 (p = 0.0167), and day 83 (p = 0.0167). Data are presented as mean ± SEM in panels (E), and (F) and compared using a two-tailed Mann-Whitney test (*p < 0.05; **p < 0.01; ***p < 0.001, ****p < 0.0001).

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