Journal: Cancer Immunology, Immunotherapy : CII

Article Title: A CD3-bispecific molecule targeting P-cadherin demonstrates T cell-mediated regression of established solid tumors in mice

doi: 10.1007/s00262-017-2081-0

Figure Lengend Snippet: PF-06671008 binds to CD3 and P-cadherin and directs T cell cytotoxicity to P-cadherin-expressing cells. a Human CD3+ T lymphocytes and b P-cadherin-expressing (triangle) or non-expressing CHO cells (circle) were co-incubated with increasing concentrations of PF-06671008 and detected with APC-labeled anti-human IgG-Fc secondary by FACS. MFI values were plotted versus the PF-06671008 concentration Log10. c P-cadherin-expressing and non-expressing CHO cells engineered to express firefly luciferase were mixed with expanded human CD3+ lymphocytes with increasing concentrations of PF-06671008 or control. Percent cytotoxicity of the P-cadherin-expressing CHO cells (triangle) and parental CHO cells (circle) was plotted against PF-06671008 concentration Log10

Article Snippet: Staining antibodies include anti-granzyme B clone 11F1 (Novocastra), anti-CD3 clone SP162 (Spring Bioscience), and anti-P-cadherin clone 56 (BD Bioscience) conjugated with the Alexa Fluor 488 protein labeling kit (Life Technologies), anti-CD4 clone EP204 (Cell Marque), anti-CD8 clone SP16 (Spring Bioscience), and anti-cleaved caspase 3 clone 5AE1 (Cell Signaling Technologies).

Techniques: Expressing, Incubation, Labeling, Concentration Assay, Luciferase

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: A CD3-bispecific molecule targeting P-cadherin demonstrates T cell-mediated regression of established solid tumors in mice

doi: 10.1007/s00262-017-2081-0

Figure Lengend Snippet: PF-06671008 directs T cell cytotoxicity and activation in response to human tumor cells dependent upon P-cadherin expression. a A panel of human tumor cell lines were used as cytotoxicity targets to determine EC50 for cytotoxicity and assessed for human P-cadherin receptor expression by quantitative FACS. The Log10 average EC50 for cytotoxicity was plotted in a linear regression curve fit versus the Log10 average ABC value for P-cadherin binding. Histogram plots of b CD69 and c 4-1BB expression by the CD3+ CD56− subset of PBMC are shown. Dose–response curves of d percent CD69 positive and e percent 4-1BB positive of CD3+ CD4+ (triangle) or CD3+ CD8+ (circle) PBMC are shown

Article Snippet: Staining antibodies include anti-granzyme B clone 11F1 (Novocastra), anti-CD3 clone SP162 (Spring Bioscience), and anti-P-cadherin clone 56 (BD Bioscience) conjugated with the Alexa Fluor 488 protein labeling kit (Life Technologies), anti-CD4 clone EP204 (Cell Marque), anti-CD8 clone SP16 (Spring Bioscience), and anti-cleaved caspase 3 clone 5AE1 (Cell Signaling Technologies).

Techniques: Activation Assay, Expressing, Binding Assay

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: A CD3-bispecific molecule targeting P-cadherin demonstrates T cell-mediated regression of established solid tumors in mice

doi: 10.1007/s00262-017-2081-0

Figure Lengend Snippet: PF-06671008 directs P-cadherin-dependent growth inhibition to established tumors in human PBMC-engrafted NSG mice. a HCT116, b SW-480, c Ls174T, and d SW620 xenograft tumors were subcutaneously implanted to NSG mice engrafted with human PBMC. Animals were weekly dosed with vehicle (circle), 0.5 mg/kg control LP-DART (square), 0.05 mg/kg PF-06671008 (vertical triangle), or 0.5 mg/kg PF-06671008 (inverted triangle). The average tumor volume (mm3 ± SEM) is plotted against the time in days post-tumor implant

Article Snippet: Staining antibodies include anti-granzyme B clone 11F1 (Novocastra), anti-CD3 clone SP162 (Spring Bioscience), and anti-P-cadherin clone 56 (BD Bioscience) conjugated with the Alexa Fluor 488 protein labeling kit (Life Technologies), anti-CD4 clone EP204 (Cell Marque), anti-CD8 clone SP16 (Spring Bioscience), and anti-cleaved caspase 3 clone 5AE1 (Cell Signaling Technologies).

Techniques: Inhibition

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: A CD3-bispecific molecule targeting P-cadherin demonstrates T cell-mediated regression of established solid tumors in mice

doi: 10.1007/s00262-017-2081-0

Figure Lengend Snippet: IHC illustrates the formation of the PF-06671008-mediated immune synapse and tumor killing. a, b HCT116 or c, d SUM149 tumor samples were examined using IHC at 6 and 7 days, respectively, following treatment with PF-06671008 or vehicle control. a Granzyme B/CD3/P-cadherin triple IHC demonstrates the relationship between P-cadherin + tumor cells (green), CD3+ infiltrating T lymphocytes (pink), and expression of Granzyme B (brown) in a PF-06671008-treated HCT116 tumor. Granzyme B+ CD3+ tumor-infiltrating lymphocytes are shown surrounded by P-cadherin+ tumor cells (insets). b Granzyme B/CD8/CD4 triple stain shows Granzyme B expression (brown) in CD8+ (green) and CD4+ (pink) cells in a PF-06671008- treated HCT116 tumor. c Cleaved Caspase-3 IHC, and d H&E staining of 0.5 mg/kg PF-06671008-treated (left panels) or control-treated (right panels) SUM149 tumors demonstrating increased cell death in response to treatment with PF-06671008. a Micron bars 200 µM, b–d 100 µM

Article Snippet: Staining antibodies include anti-granzyme B clone 11F1 (Novocastra), anti-CD3 clone SP162 (Spring Bioscience), and anti-P-cadherin clone 56 (BD Bioscience) conjugated with the Alexa Fluor 488 protein labeling kit (Life Technologies), anti-CD4 clone EP204 (Cell Marque), anti-CD8 clone SP16 (Spring Bioscience), and anti-cleaved caspase 3 clone 5AE1 (Cell Signaling Technologies).

Techniques: Expressing, Staining