Additional file 4: of Characterization of immune responses to anti-PD-1 mono and combination immunotherapy in hematopoietic humanized mice implanted with tumor xenografts

Figure S1. Experimental timeline for generation of PDX/cell line-implanted hu-CB-BRGS mice. Figure S2. Expression of hPD-L1 on MDA-MB-231 TNBC cell line. MDA-MB-231 cells grown in culture were collected and stained for expression of human PD-L1. Figure S3. Human chimerism and cell numbers in TNBC MDA-MB-231 cell line implanted hu-CB-BRGS mice harvested at d11 and d21 post treatment. a, Equivalent human hematopoietic (left), T (middle, gate: hCD45+) and CD8 (right, gate: hCD45+,CD3+) chimerism in blood of humanized mice among experimental (d11 or d21 harvest, control (−) or anti-PD-1 treatment (+) groups. b, Human hematopoietic (hCD45+) and T (CD3+) cell numbers in lymph organs of TNBC-bearing hu-CB-BRGS mice at harvest. Figure S4. Immunohistochemistry analysis of human and mouse chimerism in TNBC MDA-MB-231 cell line implanted hu-CB-BRGS mice. a, Representative IHC slides from untreated and nivolumab-treated MDA-MB-231 tumors explanted from hu-CB-BRGS mice 11 or 21 days after start of treatment. b, Increased human T-cell (CD3) densities in tumors of hu-CB-BRGS mice treated with nivolumab for 21 days. Figure S5. Expression of CD25 (clone M-A251) on FoxP3+ CD4+ and CD8+ T cells (hCD45 + CD3+) in LN and spleens of hu-CB-BRGS mice. a, Representative flow cytometry staining and b, cumulative data showing percentage of FoxP3+ T cells (left) and percentage of CD25+ among the FoxP3+ T cells (right). Figure S6. Individual data points and expression of hPD-L1 on MDA-MB-231 TNBC cell line harvested from hu-CB-BRGS mice. a, Tumor growth curves of untreated (black), nivolumab-treated (red), OKI-179-treated (green) and combination (red) of the TNBC hu-CB-BRGS mice. b, Tumors were identified as mCD45-, hCD45-, Epcam+ or HLA-A,B,C+. Figure S7. Increased detection of human T cells in IHC sections from nivolumab-treated MSI-H PDX relative to untreated MSI-H PDX or nivolumab-treated MSS PDX. (PPTX 16200 kb)