Journal: Journal of proteome research

Article Title: Bisphenol A Activates an Innate Viral Immune Response Pathway

doi: 10.1021/acs.jproteome.9b00548

Figure Lengend Snippet: CD4+T cells isolated from BPA exposed mice were analyzed by label free proteomics and pathway analysis. Differential expression is displayed as a volcano plot using log2(fold change) of BPA treated mothers (F0) by giving 10ug/ml BPA in 1% ethanol drinking water versus control (1% ethanol drinking water) out to three generations (F1-F3).

Article Snippet: CD4 + T cells were isolated from splenic mononuclear cells using magnetic beads for CD4 negative selection (StemCell, Vancouver, BC Canada).

Techniques: Isolation, Quantitative Proteomics, Control

Journal: Cancer immunology research

Article Title: A CD40 agonist and PD-1 antagonist antibody reprogram the microenvironment of non-immunogenic tumors to allow T cell–mediated anticancer activity

doi: 10.1158/2326-6066.CIR-18-0061

Figure Lengend Snippet: A) Schematic. Mice are inoculated with tumor cells on day X, specified below, and treated with vaccine on Day 0, followed by anti–PD-1 and/or CD40 agonist mAb, or rat IgG isotype control, on Day 2. Anti–PD-1 therapy is continued twice weekly thereafter, until subcutaneous tumors reach >500 mm3, or mice succumb to liver metastases (n = 10 mice per group). B) Tumor volume (n = 10 mice per group) and C) overall survival (n = 19–29 mice per group, pooled from 3 independent experiments) of neu-N mice given 5×104 NT2.5 cells in the mammary fat pad on Day X = −3 and treated as indicated. D) Tumor volume and E) overall survival of neu-N mice given 5×104 NT2.5 cells in the mammary fat pad on Day X = −3, followed by 2×106 neu-specific CD8+ T cells on Day 1, and treated as indicated. F) Overall survival of C57BL/6 mice given 2×106 PANC02 tumor cells via intrasplenic injection followed by hemisplenectomy on Day X = −15 and treated as indicated. G) Overall survival of C57BL/6 mice given 1×106 PANC02 tumor cells subcutaneously in the left lower limb on Day X = −15 and treated as indicated. *, p<0.05, **, p<0.01, ***, p<0.001, ****, p<0.0001.

Article Snippet: For adoptive T cell transfer, CD8 + T cells were isolated using the mouse CD8 negative selection kit according to the manufacturer specifications (Stem Cell Technologies), washed twice with PBS, and resuspended in PBS at 1–2×10 7 /ml for intravenous (i.v.) injection.

Techniques: Control, Injection

Journal: Cancer immunology research

Article Title: A CD40 agonist and PD-1 antagonist antibody reprogram the microenvironment of non-immunogenic tumors to allow T cell–mediated anticancer activity

doi: 10.1158/2326-6066.CIR-18-0061

Figure Lengend Snippet: A) Tumor infiltration of Thy1.2+ CD8+ T cells as measured by number of cells per mg tumor and B) CD8+ T cell/Treg ratio as measured by number of CD8+ T cells / number of CD4+ FoxP3+ T cells in tumors of neu-N mice (n = 20–24 mice per group, pooled from 6 independent experiments). Cytokine expression of CD8+ T cells from neu-N mice following stimulation of C) TIL and D) TDLN cells with RNEU peptide-pulsed APCs (n = 3–4 mice per group). E) Liver infiltration of CD8+, CD4+ Th and CD4+ Treg T cells as measured by number of cells per liver in PANC02 metastatic tumor-bearing mice. F) Cytokine expression of CD8+ T cells following stimulation of PANC02 liver TIL with CD3/CD28 activation beads. G) Overall survival of PANC02 subcutaneous tumor injected mice treated with depleting mAbs against CD8α, CD4, or isotype control. *, p<0.05, **, p<0.01, ***, p<0.001, ****, p<0.0001. TDLN=tumor draining lymph node; APCs=antigen presenting cells.

Article Snippet: For adoptive T cell transfer, CD8 + T cells were isolated using the mouse CD8 negative selection kit according to the manufacturer specifications (Stem Cell Technologies), washed twice with PBS, and resuspended in PBS at 1–2×10 7 /ml for intravenous (i.v.) injection.

Techniques: Expressing, Activation Assay, Injection, Control

Journal: Cancer immunology research

Article Title: A CD40 agonist and PD-1 antagonist antibody reprogram the microenvironment of non-immunogenic tumors to allow T cell–mediated anticancer activity

doi: 10.1158/2326-6066.CIR-18-0061

Figure Lengend Snippet: Tumor infiltration of A) CD4+ and B) CD8+ memory T cells as measured by number of cells per mg tumor, and C) CD4+ and D) CD8+ memory T cells in TDLN as measured by number of cells per LN on Day 14 in neu-N mice. E) Tumor volume and F) overall survival of triple therapy treated neu-N mice rechallenged with NT2.5 tumor cells on Day 28. Neu-N mice were inoculated with 1×106 tumor cells on Day −8 and treated with 3T3neuGM on Day 0 and 14, as well as 6×106 neu-specific T cells on Day 1. MAb therapy was administered as described until Day 28, at which point therapy was discontinued and tumor-free triple therapy treated mice, or naïve controls, were rechallenged with 5×104 NT2.5 cells in the contralateral mammary fat pad. Graphs represent days post injection of rechallenge tumor. G) Tumor volume and H) overall survival of triple therapy treated mice rechallenged with PANC02 tumors cells in the contralateral lower limb (or naïve controls) on day 60. Graphs represent days post injection of rechallenge tumor **p<0.01, ****, p<0.0001.

Article Snippet: For adoptive T cell transfer, CD8 + T cells were isolated using the mouse CD8 negative selection kit according to the manufacturer specifications (Stem Cell Technologies), washed twice with PBS, and resuspended in PBS at 1–2×10 7 /ml for intravenous (i.v.) injection.

Techniques: Injection

Journal: Cancer immunology research

Article Title: A CD40 agonist and PD-1 antagonist antibody reprogram the microenvironment of non-immunogenic tumors to allow T cell–mediated anticancer activity

doi: 10.1158/2326-6066.CIR-18-0061

Figure Lengend Snippet: A) t-SNE plots of T cell clusters from tumors of neu-N mice, separated by treatment group. Samples were gated on Live/CD3+/CD4+CD8− or Live/CD3+/CD8+CD4− T cells (n = 4 mice per group). Key clusters are labeled with T cell phenotype and cluster number. B) High dimensional flow plots showing MFI of each T cell marker, with clusters highlighted according to cluster color and overlaid on total T cells. Dot plots show frequency of each cluster per treatment group, represented by percent of live T cells. *, p<0.05, **, p<0.01, ***, p<0.001, ****, p<0.0001. t-SNE = t-distributed stochastic neighbor embedding. MFI = mean fluorescence intensity.

Article Snippet: For adoptive T cell transfer, CD8 + T cells were isolated using the mouse CD8 negative selection kit according to the manufacturer specifications (Stem Cell Technologies), washed twice with PBS, and resuspended in PBS at 1–2×10 7 /ml for intravenous (i.v.) injection.

Techniques: Labeling, Marker, Fluorescence

Journal: Cancer immunology research

Article Title: A CD40 agonist and PD-1 antagonist antibody reprogram the microenvironment of non-immunogenic tumors to allow T cell–mediated anticancer activity

doi: 10.1158/2326-6066.CIR-18-0061

Figure Lengend Snippet: PD-1 and Tim3 expression in A) Thy1.2+ CD8+ and B) Thy1.2– CD8+ T cells on Day 4, 6 and 8 from tumors of treated neu-N mice, represented as frequency of parent population (Thy1.2+ CD8+ T cells or Thy1.2− CD8+ T cells, respectively).

Article Snippet: For adoptive T cell transfer, CD8 + T cells were isolated using the mouse CD8 negative selection kit according to the manufacturer specifications (Stem Cell Technologies), washed twice with PBS, and resuspended in PBS at 1–2×10 7 /ml for intravenous (i.v.) injection.

Techniques: Expressing

Journal: Cancer immunology research

Article Title: A CD40 agonist and PD-1 antagonist antibody reprogram the microenvironment of non-immunogenic tumors to allow T cell–mediated anticancer activity

doi: 10.1158/2326-6066.CIR-18-0061

Figure Lengend Snippet: Tumor infiltration of A) CD11b+ F4/80– MDSCS (Ly6G+ G-MDSCs or Ly6C+ M-MDSCs), B) CD11b+ F4/80+ macrophages, and C) CD8+ CD11c+ MHCII+ cDCs as measured by number of cells per mg tumor in neu-N mice. D) Expression of arginase 1 (Arg1) in tumor infiltrating M-MDSCs and macrophages, represented as percent of parent population. E) Expression of CD80 and CD86 in TDLN CD11c+ cDCs, represented as number of positive cells per TDLN. F) CD103+ DCs, represented as number of positive cells per TDLN. G) Expression of CD80 and CD86 in TDLN macrophages (CD11b+ F4/80+), represented as number of positive cells per TDLN. H) CD169+ macrophages, represented as number of positive cells per TDLN. *, p<0.05, **, p<0.01, ***, p<0.001, ****, p<0.0001.

Article Snippet: For adoptive T cell transfer, CD8 + T cells were isolated using the mouse CD8 negative selection kit according to the manufacturer specifications (Stem Cell Technologies), washed twice with PBS, and resuspended in PBS at 1–2×10 7 /ml for intravenous (i.v.) injection.

Techniques: Expressing