Journal: Journal for Immunotherapy of Cancer

Article Title: Combination of gemcitabine and anti-PD-1 antibody enhances the anticancer effect of M1 macrophages and the Th1 response in a murine model of pancreatic cancer liver metastasis

doi: 10.1136/jitc-2020-001367

Figure Lengend Snippet: The response in pancreatic ductular adenocarcinoma (PDAC) liver metastasis tumors of mice treated with gemcitabine (GEM) and anti-programmed cell death 1 (anti-PD-1) Ab. (A) Experimental schedule showing the treatments for PDAC liver metastasis mice: no treatment (phosphate-buffered saline), anti-PD-1 Ab, GEM, and GEM plus anti-PD-1 Ab. Treatment was administered twice a week from day 7 to day 33. Tumor tissues were obtained and stained immunohistochemically on day 34 (n=3 per group). (B) Number of liver metastasis tumor nodules and averaged nodule volumes and (C) macroscopic images of tumors are shown; (B) bars represent mean±SEM; Student’s t-test was performed as statistical analysis. (D) Immunohistochemical analysis of tumors for CD8a+, CD4+, and CD11b+ inflammatory cells. Magnification: ×200; bars: 100 µm. Quantification of cell infiltration by using ImageJ (each area analyzed=1.576 mm 2 , three different areas were analyzed for each sample). White bar: no treatment; light gray bar: anti-PD-1 Ab; dark gray bar: GEM; black bar: GEM plus anti-PD-1 Ab; bars represent mean±SEM; one-way analysis of variance followed by Tukey’s HSD post hoc test was performed as statistical analysis; *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001. DAB, 3,3'-diaminobenzidine; HSD, honestly significant difference.

Article Snippet: The PDAC mice were treated with 50 mg/kg GEM (G0367; TCI, Tokyo, Japan) by tail vein injection, and with or without intraperitoneal injection of anti-PD-1 Ab (200 µg; clone: J43; BD Biosciences, Franklin Lakes, New Jersey) or anti-CD8 Ab (200 µg; clone: 53–6.7; BD Biosciences).

Techniques: Staining, Immunohistochemical staining

Journal: Journal for Immunotherapy of Cancer

Article Title: Combination of gemcitabine and anti-PD-1 antibody enhances the anticancer effect of M1 macrophages and the Th1 response in a murine model of pancreatic cancer liver metastasis

doi: 10.1136/jitc-2020-001367

Figure Lengend Snippet: Gene expression analysis of tumor-infiltrating inflammatory cells (TICs) from pancreatic ductular adenocarcinoma (PDAC) liver metastasis model mice by qRT-PCR. PDAC liver metastasis mice received a single dose of the indicated treatment on day 28 and tumor tissues were obtained 2 days later for TIC isolation, followed by RNA extraction. Quantitative RT-PCR showing the expression of M1 macrophage-related cytokine genes ( Il6 , Il12a , Il12b , Il1b , and Tnf ), M2 macrophage-related genes ( Arg1 , Il10 , Tgfb1 , and Mmp9 ), proinflammatory chemokines ( Cxcl10 and Ccl2 ), and T-cell activation markers ( Pdcd1 and Prf1 ); n=3; bars represent mean±SEM; Student’s t-test was used for statistical analysis; *p<0.05, **p<0.01. GEM, gemcitabine; PD-1, programmed cell death 1; qRT-PCR, quantitative real-time polymerase chain reaction.

Article Snippet: The PDAC mice were treated with 50 mg/kg GEM (G0367; TCI, Tokyo, Japan) by tail vein injection, and with or without intraperitoneal injection of anti-PD-1 Ab (200 µg; clone: J43; BD Biosciences, Franklin Lakes, New Jersey) or anti-CD8 Ab (200 µg; clone: 53–6.7; BD Biosciences).

Techniques: Expressing, Quantitative RT-PCR, Isolation, RNA Extraction, Activation Assay, Real-time Polymerase Chain Reaction

Journal: Journal for Immunotherapy of Cancer

Article Title: Combination of gemcitabine and anti-PD-1 antibody enhances the anticancer effect of M1 macrophages and the Th1 response in a murine model of pancreatic cancer liver metastasis

doi: 10.1136/jitc-2020-001367

Figure Lengend Snippet: Flow cytometry (FCM) analysis of tumor-infiltrating inflammatory cells (TICs) for lymphoid-lineage cells. Pancreatic ductular adenocarcinoma liver metastasis mice received a single dose of the indicated treatment on day 28 and tumor tissues were obtained 2 days later for TIC isolation followed by FCM analysis; n=3. (A) CD4+ and CD8a+ cells within TICs. (B) Th1 CD4+ cells expressing T-bet and interferon (IFN)-γ; IFN-γ-secreting cells within CD8+ TICs. (C) Regulatory T cell phenotype, CD4+CD25+FoxP3+ cells, and interleukin (IL)-10+-producing cells within Treg cells. (A–C) Bars represent mean±SEM; one-way analysis of variance followed by Tukey’s HSD post hoc test was performed as statistical analysis; *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001. GEM, gemcitabine. PD-1, programmed cell death 1.

Article Snippet: The PDAC mice were treated with 50 mg/kg GEM (G0367; TCI, Tokyo, Japan) by tail vein injection, and with or without intraperitoneal injection of anti-PD-1 Ab (200 µg; clone: J43; BD Biosciences, Franklin Lakes, New Jersey) or anti-CD8 Ab (200 µg; clone: 53–6.7; BD Biosciences).

Techniques: Flow Cytometry, Isolation, Expressing

Journal: Journal for Immunotherapy of Cancer

Article Title: Combination of gemcitabine and anti-PD-1 antibody enhances the anticancer effect of M1 macrophages and the Th1 response in a murine model of pancreatic cancer liver metastasis

doi: 10.1136/jitc-2020-001367

Figure Lengend Snippet: Immunohistochemical analysis of pancreatic ductular adenocarcinoma liver metastasis tumors for myeloid-lineage cells. Treatments were conducted twice a week from day 7 to day 33; tumors were obtained on day 34 and immunohistochemically analyzed for (A) F4/80+, Ly6C+, and Ly6G+, and (B) CD86+, CD206+ infiltrating cells, and programmed cell death-ligand 1 (PD-L1)+ for the indicated treatment. Magnification: ×200; bars: 100 µm. (A, B) Quantification of cell infiltration by using ImageJ (each area analyzed=1.576 mm 2 , 3 different areas were analyzed for each sample). White bar: no treatment; light gray bar: anti-PD-1 Ab; dark gray bar: gemcitabine (GEM); black bar: GEM plus anti-PD-1 Ab. Bars represent mean±SEM; one-way analysis of variance followed by Tukey’s HSD post hoc test was performed as statistical analysis; *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001.

Article Snippet: The PDAC mice were treated with 50 mg/kg GEM (G0367; TCI, Tokyo, Japan) by tail vein injection, and with or without intraperitoneal injection of anti-PD-1 Ab (200 µg; clone: J43; BD Biosciences, Franklin Lakes, New Jersey) or anti-CD8 Ab (200 µg; clone: 53–6.7; BD Biosciences).

Techniques: Immunohistochemical staining

Journal: Journal for Immunotherapy of Cancer

Article Title: Combination of gemcitabine and anti-PD-1 antibody enhances the anticancer effect of M1 macrophages and the Th1 response in a murine model of pancreatic cancer liver metastasis

doi: 10.1136/jitc-2020-001367

Figure Lengend Snippet: Flow cytometry (FCM) analysis of tumor-infiltrating inflammatory cells (TICs) for myeloid-lineage cells. Pancreatic ductular adenocarcinoma liver metastasis mice received the following treatments: no treatment, anti-programmed cell death 1 (anti-PD-1) Ab, gemcitabine (GEM), or GEM plus anti-PD-1 Ab on day 28 and tumor tissues were obtained 2 days later for TIC isolation and FCM; n=3, bars represent mean±SEM. The following cell populations were analyzed: Ly6C+Ly6G− inflammatory monocytes within CD11b+F4/80+ TICs, CD206− M1 macrophages within CD11b+F4/80 high TICs, and CD206+ M2 macrophages within CD11b+F4/80 high TICs. One-way analysis of variance followed by Tukey’s HSD post hoc test was performed as statistical analysis; *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001.

Article Snippet: The PDAC mice were treated with 50 mg/kg GEM (G0367; TCI, Tokyo, Japan) by tail vein injection, and with or without intraperitoneal injection of anti-PD-1 Ab (200 µg; clone: J43; BD Biosciences, Franklin Lakes, New Jersey) or anti-CD8 Ab (200 µg; clone: 53–6.7; BD Biosciences).

Techniques: Flow Cytometry, Isolation

Journal: Journal for Immunotherapy of Cancer

Article Title: Combination of gemcitabine and anti-PD-1 antibody enhances the anticancer effect of M1 macrophages and the Th1 response in a murine model of pancreatic cancer liver metastasis

doi: 10.1136/jitc-2020-001367

Figure Lengend Snippet: Survival of pancreatic ductular adenocarcinoma (PDAC) liver metastasis model mice that underwent treatment. (A) Experimental schedule showing treatment timing: the PDAC liver metastasis mice received treatment 10 times, twice a week from day 7 to day 39, and survival percentage was monitored until day 72. (B) Survival curves of PDAC liver metastasis mice that received: no treatment (only phosphate-buffered saline; n=7), anti-programmed cell death 1 (anti-PD-1) Ab (n=12), gemcitabine (GEM) (n=7), GEM plus anti-PD-1 Ab (n=6), or GEM plus anti-PD-1 Ab plus anti-CD8 Ab (n=7). The log-rank test was performed to obtain the p values: no treatment/anti-PD-1 Ab=0.492723; no treatment/GEM=0.000172; no treatment/GEM plus anti-PD-1 Ab=0.000446; anti-PD-1 Ab/GEM=0.000034; anti-PD-1 Ab/GEM plus anti-PD-1 Ab=0.000098; GEM/GEM plus anti-PD-1 Ab=0.034672; and GEM plus anti-PD-1 Ab/GEM plus anti-PD-1 Ab plus anti-CD8 Ab=0.0345.

Article Snippet: The PDAC mice were treated with 50 mg/kg GEM (G0367; TCI, Tokyo, Japan) by tail vein injection, and with or without intraperitoneal injection of anti-PD-1 Ab (200 µg; clone: J43; BD Biosciences, Franklin Lakes, New Jersey) or anti-CD8 Ab (200 µg; clone: 53–6.7; BD Biosciences).

Techniques:

Journal: Journal for Immunotherapy of Cancer

Article Title: Combination of gemcitabine and anti-PD-1 antibody enhances the anticancer effect of M1 macrophages and the Th1 response in a murine model of pancreatic cancer liver metastasis

doi: 10.1136/jitc-2020-001367

Figure Lengend Snippet: In vitro macrophage polarization assay. Pancreatic ductular adenocarcinoma liver metastasis mice received a single dose of the indicated treatment on day 28 and splenocytes were obtained on day 30. The splenocytes were activated by coculture with irradiated PAN02 cells; after 7 days, activated splenocytes were harvested and cocultured with bone marrow-derived macrophages. After 48 hours, macrophages were collected for (A) flow cytometry analysis of CD11b, CD86, and CD206; and total RNA was isolated for qRT-PCR analysis of (B) Il6 , Il12a , Il1b , and Tnf (M1 macrophage-related genes) or (C) Arg1 , Il10 , Tgfb1 , and Mmp9 (M2 macrophage-related genes). (A–C) n=3; bars represent mean±SEM; Student’s t-test was performed for statistical analysis; *p<0.05, **p<0.01, ***p<0.001. GEM, gemcitabine; PD-1, programmed cell death 1.

Article Snippet: The PDAC mice were treated with 50 mg/kg GEM (G0367; TCI, Tokyo, Japan) by tail vein injection, and with or without intraperitoneal injection of anti-PD-1 Ab (200 µg; clone: J43; BD Biosciences, Franklin Lakes, New Jersey) or anti-CD8 Ab (200 µg; clone: 53–6.7; BD Biosciences).

Techniques: In Vitro, Irradiation, Derivative Assay, Flow Cytometry, Isolation, Quantitative RT-PCR

Journal: Otolaryngology--Head and Neck Surgery

Article Title: PD‐L1 Acts Independently of PD‐1 as a Marker of Pathologic Fibroblasts in Laryngotracheal Stenosis

doi: 10.1002/ohn.1034

Figure Lengend Snippet: PD‐1 KO mice are not protected from laryngotracheal stenosis. (A) Significant increase in lamina propria thickness following injury with no significant difference between PD1 KO and WT. (B) Sample sections. Scale bar = 200 µm. KO, knockout; ns, not significant; PD‐1, programmed cell death 1; WT, wild type. ** P < .01; *** P < .001.

Article Snippet: Matched scar and healthy fibroblasts from 1 iSGS patient were also cultured in 24‐well plates that were untreated or precoated with 5 µg/mL of the PD‐1 agonist, PD‐1‐Fc‐Fusion (AB Biosciences), or Isotype control IgG2b (Biolegend).

Techniques: Knock-Out

Journal: Cancer Management and Research

Article Title: Epigenome-Driven Strategies for Personalized Cancer Immunotherapy

doi: 10.2147/CMAR.S272031

Figure Lengend Snippet: Some Immunotherapy Treatments, Their Associated Epigenetic Hallmarks Found in Cancer Patients and Clinical Outcomes

Article Snippet: NSCLC , Use of anti-PD-1 monoclonal Ab , FOXP1 demethylation and EPIMMUNE epigenetic signature , Clinical benefit with PD-1 blockade , [ ] .

Techniques: Methylation, Disruption, Activity Assay

Journal: Cancers

Article Title: Protein Expression in Metastatic Melanoma and the Link to Disease Presentation in a Range of Tumor Phenotypes

doi: 10.3390/cancers12030767

Figure Lengend Snippet: Combinative treatments for metastatic melanoma.

Article Snippet: , Anti PD-1 ab (from BioXCell) LBH589 (HDACi) , Slower tumor progression and increased survival compared with control and single treatments in B16F10 mice model , [ ] .

Techniques: Control