Figure 4.
Immune Reconstitution. (A) Lymphocyte repopulation. Time course of lymphocyte subsets in AC (blue) and TMS (red) treatment arm patients at 0.5, 1, 3, 6, 12, and 24 months. Medians are indicated as circles, 25th and 75th quartiles are shown as error bars. Circles placed below the x-axis indicate a median of 0 cells. For patient numbers and statistical disparity between arms at each T-cell time point, refer to supplemental Figure 5A. (B) Time course of changes in posttransplant TCR repertoire skewing. Repertoire skewing (oligoclonality) indices (RSI) determined for individual AC (blue circles) and TMS (red circles) patients at 1, 3, 6, and 12 months were compared between arms (top statistics) and with the RSI of their own allogeneic transplant donors (green triangles) (bottom statistics). Both CD4 (left graph) and CD8 cells (right graph) in the AC arm had more repertoire skewing (more oligoclonality) than those in most patients in the TMS arm or in their donor’s original T cells at 1, 3, and 6 months in CD4 and at 3 and 6 months in CD8 T cells. (C) Flow cytometry gating profiles of T lymphocytes at 6 months in the AC and TMS arms. CD4 cells were gated to distinguish Treg (thick outline) from non-Treg (conventional) CD4 T cells, based on Treg characterization as CD127–CD25++ CD4 cells. The non-Treg and Treg CD4 and the CD8 cells were then gated to assess naïve (CD45RA+CCR7+; thick outline), central memory (CD45RA–CCR7+), effector memory (CD45RA–CCR7–) and TEMRA (CD45RA+CCR7–) subsets. (D) Box and whisker plots of the percentage of Treg cells within the CD4 population (left graph) and the total number of Treg per μL (right graph) in AC (blue box) and TMS (orange box) patients at 6, 12, and 24 months after transplant. (E) Box and whisker plots of the numbers of naïve CD4 T cells per μL (left graph) and of naïve CD8 T cells per μL (right graph) at 6, 12, and 24 months. (F) Box and whisker plots of the ratios of the numbers of Treg to the number of naïve CD4 (left graph) or to the number of naïve CD8 T cells (right graph) at 6, 12, and 24 months. (G) Flow cytometry gating profiles of CD8 Tscm populations. TMS patient flow cytometry panels identifying naïve (CD45RA+ CCR7+) CD8 cells (thick line box) at 6, 9, and 12 months (left column), showing gating on the CD8naïve (arrows) to identify CCR7dimCD95+ Tscm at 3 sequential time points (right column). The patient shown developed moderate cGVHD at 9 months. (H) Box and whiskers plot comparing the number of CD8 Tscm per μL in AC (blue box) and TMS (orange box) at 6, 12, and 24 months. (I) Scatterplot of individual AC (blue circle) and TMS (red circle) patients as assessed for the ratio of Treg per μL to CD8naïve per μL vs the number of Tscm per μL. In all box and whisker plots, the number of patients assayed at each time point in each arm is shown in supplemental Figure 5C. In all graphs, AC arm patients are shown in blue box and TMS in orange box; boxes define the median, 25th and 75th quartiles; and whiskers show minimum and maximum values. Mann-Whitney unpaired nonparametric statistics were performed to compare AC and TMS lymphocyte numbers, RSI, and lymphocyte subpopulations and ratios; shown as stars: ∗∗∗∗P < .0001; ∗∗∗P < .001; ∗∗P < .01; ∗ P < .05. NK cell, natural killer cell.

Immune Reconstitution. (A) Lymphocyte repopulation. Time course of lymphocyte subsets in AC (blue) and TMS (red) treatment arm patients at 0.5, 1, 3, 6, 12, and 24 months. Medians are indicated as circles, 25th and 75th quartiles are shown as error bars. Circles placed below the x-axis indicate a median of 0 cells. For patient numbers and statistical disparity between arms at each T-cell time point, refer to supplemental Figure 5A. (B) Time course of changes in posttransplant TCR repertoire skewing. Repertoire skewing (oligoclonality) indices (RSI) determined for individual AC (blue circles) and TMS (red circles) patients at 1, 3, 6, and 12 months were compared between arms (top statistics) and with the RSI of their own allogeneic transplant donors (green triangles) (bottom statistics). Both CD4 (left graph) and CD8 cells (right graph) in the AC arm had more repertoire skewing (more oligoclonality) than those in most patients in the TMS arm or in their donor’s original T cells at 1, 3, and 6 months in CD4 and at 3 and 6 months in CD8 T cells. (C) Flow cytometry gating profiles of T lymphocytes at 6 months in the AC and TMS arms. CD4 cells were gated to distinguish Treg (thick outline) from non-Treg (conventional) CD4 T cells, based on Treg characterization as CD127–CD25++ CD4 cells. The non-Treg and Treg CD4 and the CD8 cells were then gated to assess naïve (CD45RA+CCR7+; thick outline), central memory (CD45RA–CCR7+), effector memory (CD45RA–CCR7–) and TEMRA (CD45RA+CCR7–) subsets. (D) Box and whisker plots of the percentage of Treg cells within the CD4 population (left graph) and the total number of Treg per μL (right graph) in AC (blue box) and TMS (orange box) patients at 6, 12, and 24 months after transplant. (E) Box and whisker plots of the numbers of naïve CD4 T cells per μL (left graph) and of naïve CD8 T cells per μL (right graph) at 6, 12, and 24 months. (F) Box and whisker plots of the ratios of the numbers of Treg to the number of naïve CD4 (left graph) or to the number of naïve CD8 T cells (right graph) at 6, 12, and 24 months. (G) Flow cytometry gating profiles of CD8 Tscm populations. TMS patient flow cytometry panels identifying naïve (CD45RA+ CCR7+) CD8 cells (thick line box) at 6, 9, and 12 months (left column), showing gating on the CD8naïve (arrows) to identify CCR7dimCD95+ Tscm at 3 sequential time points (right column). The patient shown developed moderate cGVHD at 9 months. (H) Box and whiskers plot comparing the number of CD8 Tscm per μL in AC (blue box) and TMS (orange box) at 6, 12, and 24 months. (I) Scatterplot of individual AC (blue circle) and TMS (red circle) patients as assessed for the ratio of Treg per μL to CD8naïve per μL vs the number of Tscm per μL. In all box and whisker plots, the number of patients assayed at each time point in each arm is shown in supplemental Figure 5C. In all graphs, AC arm patients are shown in blue box and TMS in orange box; boxes define the median, 25th and 75th quartiles; and whiskers show minimum and maximum values. Mann-Whitney unpaired nonparametric statistics were performed to compare AC and TMS lymphocyte numbers, RSI, and lymphocyte subpopulations and ratios; shown as stars: ∗∗∗∗P < .0001; ∗∗∗P < .001; ∗∗P < .01; ∗ P < .05. NK cell, natural killer cell.

Close Modal
This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal