Figure 2
Figure 2. BM NKPs reconstitute NK and T cells in vivo. Sublethally irradiated 10- to 12-week-old Il2rg−/− CD45.2 recipient mice were transplanted with 1300 BM Lin−CD122+NK1.1−DX5− NKPs sorted from 9- to 12-week-old CD45.1 WT mice. Donor-derived lineage reconstitution was evaluated in the PB, spleen, and BM at 14, 21, and 28 days after transplantation by FACS. (A) Representative FACS plots of donor-derived CD45.1+ NK, T, B, and myeloid cell reconstitution in the PB and in the spleen from representative mice at 21 days after transplantation. (Top panels) Nontransplanted CD45.1 control mouse. (Bottom panels) Il2rg−/− transplanted recipient mouse. The specific gates are indicated in the text above the plot and by the arrow. The numbers show the proportion of cells within the gate, of total (CD45.1 plus CD45.2) cells and represent mean values from 9 to 13 mice from 2 independent experiments. (B) Donor-derived total reconstitution (CD45.1+) and donor-derived NK cells (CD45.1+CD3−NK1.1+DX5+) at 14, 21, and 28 days after transplantation in the PB. Data represent mean (SD) of total cells per 1 mL of PB; 4 to 8 mice were analyzed per group in 1 or 2 experiments. (C) Donor-derived total reconstitution (CD45.1+), donor-derived NK cells (CD45.1+CD3−NK1.1+DX5+), donor-derived T cells (CD45.1+CD4+/CD8+NK1.1−), donor-derived B cells (CD45.1+CD19+), and donor-derived myeloid cells (CD45.1+NK1.1−Mac-1+) at 21 days after transplantation in the PB. Data represent mean (SD) values of total cells per 1 mL of PB; 13 mice were analyzed per group in 2 independent experiments. (D) Donor-derived total (CD45.1+), NK (CD45.1+CD3−NK1.1+DX5+), T (CD45.1+CD4+/CD8+NK1.1−), and NK1.1+TCR+ T cells (CD45.1+CD3+NK1.1+) at 21 days after transplantation in the spleen. Data represent mean (SD) values of total cells per spleen; 9 mice were analyzed per group in 2 independent experiments. (E) Donor-derived total (CD45.1+), B cell (CD45.1+CD19+), myeloid cell (CD45.1+NK1.1−Mac-1+), and NK1.1+ cell (CD45.1+NK1.1+Mac-1−) reconstitution at 21 days after transplantation in the BM. Data represent mean (SD) values of total cells per 2 tibiae and 2 femora; 8 mice were analyzed per group. Representative FACS profiles are shown in supplemental Figure 4.

BM NKPs reconstitute NK and T cells in vivo. Sublethally irradiated 10- to 12-week-old Il2rg−/− CD45.2 recipient mice were transplanted with 1300 BM LinCD122+NK1.1DX5 NKPs sorted from 9- to 12-week-old CD45.1 WT mice. Donor-derived lineage reconstitution was evaluated in the PB, spleen, and BM at 14, 21, and 28 days after transplantation by FACS. (A) Representative FACS plots of donor-derived CD45.1+ NK, T, B, and myeloid cell reconstitution in the PB and in the spleen from representative mice at 21 days after transplantation. (Top panels) Nontransplanted CD45.1 control mouse. (Bottom panels) Il2rg−/− transplanted recipient mouse. The specific gates are indicated in the text above the plot and by the arrow. The numbers show the proportion of cells within the gate, of total (CD45.1 plus CD45.2) cells and represent mean values from 9 to 13 mice from 2 independent experiments. (B) Donor-derived total reconstitution (CD45.1+) and donor-derived NK cells (CD45.1+CD3NK1.1+DX5+) at 14, 21, and 28 days after transplantation in the PB. Data represent mean (SD) of total cells per 1 mL of PB; 4 to 8 mice were analyzed per group in 1 or 2 experiments. (C) Donor-derived total reconstitution (CD45.1+), donor-derived NK cells (CD45.1+CD3NK1.1+DX5+), donor-derived T cells (CD45.1+CD4+/CD8+NK1.1), donor-derived B cells (CD45.1+CD19+), and donor-derived myeloid cells (CD45.1+NK1.1Mac-1+) at 21 days after transplantation in the PB. Data represent mean (SD) values of total cells per 1 mL of PB; 13 mice were analyzed per group in 2 independent experiments. (D) Donor-derived total (CD45.1+), NK (CD45.1+CD3NK1.1+DX5+), T (CD45.1+CD4+/CD8+NK1.1), and NK1.1+TCR+ T cells (CD45.1+CD3+NK1.1+) at 21 days after transplantation in the spleen. Data represent mean (SD) values of total cells per spleen; 9 mice were analyzed per group in 2 independent experiments. (E) Donor-derived total (CD45.1+), B cell (CD45.1+CD19+), myeloid cell (CD45.1+NK1.1Mac-1+), and NK1.1+ cell (CD45.1+NK1.1+Mac-1) reconstitution at 21 days after transplantation in the BM. Data represent mean (SD) values of total cells per 2 tibiae and 2 femora; 8 mice were analyzed per group. Representative FACS profiles are shown in supplemental Figure 4.

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