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

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

Supplementary Methods. (DOCX 26 kb

Additional file 3: of Characterization of immune responses to anti-PD-1 mono and combination immunotherapy in hematopoietic humanized mice implanted with tumor xenografts

Table S2. Flow Cytometric Abs and Reagents. (XLSX 49 kb

Additional file 2: of Characterization of immune responses to anti-PD-1 mono and combination immunotherapy in hematopoietic humanized mice implanted with tumor xenografts

Table S1. Experimental details and timeline for tumor growth studies in BRGS and hu-CB-BRGS mice. (DOCX 20 kb

Ionic control of ? cell function in nesidioblastosis. A possible therapeutic role for calcium channel blockade

A preterm female infant presented with intractable hypoglycaemia within 10 minutes of delivery. Normoglycaemia could be maintained only by the intravenous infusion of glucose at a rate of 20-22 mg/kg/min. Persistent hyperinsulinaemic hypoglycaemia of infancy was diagnosed from an inappropriately raised plasma insulin concentration (33 mU/l) at the time of hypoglycaemia (blood glucose <0.5 mmol/l). Medical treatment with glucagon, somatostatin, and diazoxide led to only a modest reduction in the intravenous glucose requirement; a 95 pancreatectomy was performed and histological 'nesidioblastosis' confirmed. In vitro electrophysiological studies using patch clamp techniques on isolated pancreatic ? cells characterised the ionic basis for insulin secretion in nesidioblastosis. The ? cells were depolarised in low ambient glucose concentrations with persistently firing action potentials; these were blocked reversibly by the calcium channel blocking agent verapamil. Persistent postoperative hyperinsulinaemic hypoglycaemia was treated with oral nifedipine. This increased median blood glucose concentrations from 3.5 to 4.8 mmol/l and increased in duration the child's tolerance to fasting from 3 to 10.5 hours. These data allude to an abnormality in the ionic control of insulin release in nesidioblastosis and offer a new logical approach to treatment which requires further evaluation.</p