Figure 4.
Figure 4. Donor T cells competing for IL-15 signaling impairs NK-cell reconstitution. (A) B6 and B6D2F1 recipients were transplanted with B6.CD45.1+ BM and CD3+ T cells or TCD BM and donor NK cells were quantified at day 14. BM and spleen data show mean ± SEM pooled from 2 to 4 independent experiments; n = 9-18 per group. pLN data show mean ± SEM pooled from 1 to 5 independent experiments; n = 4-23 per group. (B) B6D2F1 recipients were transplanted with 5 × 106 B6.BM and 1 × 106 B6.CD3+ T cells or 0.5 × 106 sorted B6.CD4+ or 0.5 × 106 sorted B6.CD8+ T cells or B6.TCD BM. Donor NK cells were quantified at day 14. Mean ± SD from 1 experiment; n = 4 per group. (C) B6D2F1 recipients received B6.TCD BM, B6.BM or IL-21R−/−, IFNγR−/−, IFNαR−/−, TNFRI/II−/−, or NKp46cre+.TGFRIIfl/fl BM together with B6.CD3+ T cells and BM and spleen NK cells were quantified at day 14. Mean ± SEM pooled from 2 to 5 experiments for B6 TCD, B6 BM + T-cell and IFNγR−/− TCD BM groups; n = 8-20. Mean ± SD from a single experiment for remaining groups; n = 2-5 per group. (D) Quantitation of serum IL-15/IL-15R complex levels by enzyme-linked immunosorbent assay (ELISA) at day 14 posttransplant. Enumeration of respective donor NK-cell numbers in BM and spleen is shown. Mean ± SEM, pooled from 3 to 5 experiments; n = 10-18 per group. (E) Recipients were transplanted with B6 BM and either B6.CD3+ T cells or IL-15Rα−/−.CD3+ T cells. BM and T cells for these experiments originated in the United States, hence the reduced overall cell numbers. Donor NK cells were enumerated in the BM and spleen at day 18 posttransplant. Mean ± SD from 1 experiment; n = 3-6 per group. (F) B6D2F1 recipients were transplanted with donor grafts containing either B6 TCD BM with B6.CD3+ T cells, B6 TCD BM alone, or B6.CD11c-DOG TCD BM alone. CD11c+ cells were depleted with diphtheria toxin (DT) administered from day 5 post-SCT (160 ng per dose per mouse) and then every 2 days thereafter. NK cells were enumerated in BM and spleen on day 14. Mean ± SEM from 1 experiment is shown; n = 6 per group. Kruskall-Wallis test with the Dunn multiple comparisons correction was performed in panels A through C. Comparisons were made with the TCD BM group in panel C. The Mann-Whitney test was performed in panels D, E, and F.

Donor T cells competing for IL-15 signaling impairs NK-cell reconstitution. (A) B6 and B6D2F1 recipients were transplanted with B6.CD45.1+ BM and CD3+ T cells or TCD BM and donor NK cells were quantified at day 14. BM and spleen data show mean ± SEM pooled from 2 to 4 independent experiments; n = 9-18 per group. pLN data show mean ± SEM pooled from 1 to 5 independent experiments; n = 4-23 per group. (B) B6D2F1 recipients were transplanted with 5 × 106 B6.BM and 1 × 106 B6.CD3+ T cells or 0.5 × 106 sorted B6.CD4+ or 0.5 × 106 sorted B6.CD8+ T cells or B6.TCD BM. Donor NK cells were quantified at day 14. Mean ± SD from 1 experiment; n = 4 per group. (C) B6D2F1 recipients received B6.TCD BM, B6.BM or IL-21R−/−, IFNγR−/−, IFNαR−/−, TNFRI/II−/−, or NKp46cre+.TGFRIIfl/fl BM together with B6.CD3+ T cells and BM and spleen NK cells were quantified at day 14. Mean ± SEM pooled from 2 to 5 experiments for B6 TCD, B6 BM + T-cell and IFNγR−/− TCD BM groups; n = 8-20. Mean ± SD from a single experiment for remaining groups; n = 2-5 per group. (D) Quantitation of serum IL-15/IL-15R complex levels by enzyme-linked immunosorbent assay (ELISA) at day 14 posttransplant. Enumeration of respective donor NK-cell numbers in BM and spleen is shown. Mean ± SEM, pooled from 3 to 5 experiments; n = 10-18 per group. (E) Recipients were transplanted with B6 BM and either B6.CD3+ T cells or IL-15Rα−/−.CD3+ T cells. BM and T cells for these experiments originated in the United States, hence the reduced overall cell numbers. Donor NK cells were enumerated in the BM and spleen at day 18 posttransplant. Mean ± SD from 1 experiment; n = 3-6 per group. (F) B6D2F1 recipients were transplanted with donor grafts containing either B6 TCD BM with B6.CD3+ T cells, B6 TCD BM alone, or B6.CD11c-DOG TCD BM alone. CD11c+ cells were depleted with diphtheria toxin (DT) administered from day 5 post-SCT (160 ng per dose per mouse) and then every 2 days thereafter. NK cells were enumerated in BM and spleen on day 14. Mean ± SEM from 1 experiment is shown; n = 6 per group. Kruskall-Wallis test with the Dunn multiple comparisons correction was performed in panels A through C. Comparisons were made with the TCD BM group in panel C. The Mann-Whitney test was performed in panels D, E, and F.

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