Table 1.
Patient Demographics and Baseline Disease Characteristics (N = 35 patients).
Table 2.
Adverse events (worst grade) that were possibly, probably or definitely related to ipilimumab (N = 35; incidence ≥2)*.
Figure 1.
Forest plot of the multicolor flow cytometry data comparing the cell surface marker expression of regulatory T cells (Treg) and myeloid-derived suppressor cells (MDSC) on peripheral blood mononuclear cells (PBMC) at baseline and following treatment with ipilimumab (6 weeks).
The plot represents average within-patient changes from baseline to 6 weeks (with corresponding 95% confidence intervals). Treg were defined as cells expressing (1) CD4+CD25hi+FOXP3+ or (2) CD4+CD25hi+CD39+ activated T cells (CD3+CD4+CD25+). MDSC were defined as cells expressing (1) Lin1−/HLA-DR−/CD33+/CD11b+ lymphoid gate, (2) Lin1−/HLA-DR−/CD33+/CD11b+ monocyte gate or (3) HLA-DR+lo/CD14+ monocyte gate (N = 27 patients). Examples of raw data are provided in Figures S3 (T-reg gating) and S4 (MDSC gating).
Figure 2.
Kaplan-Meier plot of progression free survival (PFS) by the dichotomized (at median) change in the percentage of circulating regulatory T cells (Treg) between baseline and week 6.
Greater increase in circulating Treg (CD4+CD25hi+Foxp3+%) was associated with improved PFS (HR = 0.57, p = 0.034; N = 27 patients). Example of raw data is provided in Figure S3 where the gating strategy used for Treg is shown.
Figure 3.
Kaplan-Meier plot of progression free survival (PFS) by the dichotomized change in the percentage of circulating MDSC between baseline and week 6.
Greater decrease in circulating monocyte gate MDSC (Lin1−/HLA-DR−/CD33+/CD11b+%) was associated with improved progression free survival (PFS; p = 0.03; N = 27 patients). Example of raw data is provided in Figure S4 where the gating strategies used for MDSC subsets are shown.
Figure 4.
Immunohistochemistry (IHC) of CD8+ tumor infiltrating lymphocytes (TIL).
There was a significant increase in CD8+ T cells from baseline to week 6 (Wilcoxon signed-rank test p = 0.02; N = 24 patients). (A) Boxplots of IHC scores of CD8+ TIL at baseline and week 6. Total counts from the same patient at the two time points are connected by light gray lines. (B) Plot of the median change in CD8+ TIL at week 6 (compared to baseline) with corresponding 95% confidence intervals. (C–D) Example of baseline and week 6 tumor CD8+ TIL (stained brown) by IHC. Magnification: 20×.