Journal: Immune Network

Article Title: Adenosine Blockage in Tumor Microenvironment and Improvement of Cancer Immunotherapy

doi: 10.4110/in.2019.19.e23

Figure Lengend Snippet: Clinical trials with adenosine pathway inhibitors combined with an immune check inhibitor in cancer

Article Snippet: A2AR , PFB509 , Novartis/Palobiofarma , I, Ib , NSCLC , PDR001 (anti-PD-1) , NCT02403193.

Techniques: Clinical Proteomics

Journal: mBio

Article Title: A Combination of Anti-PD-L1 Treatment and Therapeutic Vaccination Facilitates Improved Retroviral Clearance via Reactivation of Highly Exhausted T Cells

doi: 10.1128/mBio.02121-20

Figure Lengend Snippet: Nanoparticle-based therapeutic vaccination synergizes with a PD-L1 blockade to increase retrovirus-specific CD8 + T cell immunity. (A) Chronically FV-infected mice were treated twice with αPD-L1 or an isotype control (Iso) antibody, starting at 6 weeks postinfection. Groups of mice received therapeutic vaccination with CpG and GagL 85–93 -functionalized CaP nanoparticles alone or in addition to the PD-L1 blockade at the beginning of the treatment. Seven days after the initial treatment, the CD8 + T cell response was analyzed. (B and D) Numbers of total CD8 + T cells (B) or percentages of GagL 85–93 -specific tetramer + CD8 + T cells (D) were determined in the spleen by counting viable cells using trypan blue staining. Cell counts were applied to viable-cell populations in a flow cytometric analysis. (C) Representative dot plots from flow cytometry showing the frequencies of Gag-specific tetramer + CD8 + T cells. (E) Representative histogram from flow cytometry showing GzmB-expressing CD43 + tetramer + CD8 + T cells. (F) Mean fluorescent intensity (MFI) for GzmB gated on CD43 + tetramer + CD8 + T cells. (G) Ratio between GzmB-expressing CD43 + tetramer + CD8 + T cells and Foxp3-expressing CD4 + regulatory T cells in the spleen. (H) Seven days after initial treatment, an in vivo cytotoxicity assay was performed to determine the killing capacity of Gag-specific CD8 + T cells. Representative histograms of CD45.1 gated donor cells from the spleen showing in vivo killing of target cells loaded with FV GagL peptide. (I) Elimination of donor CD45.1 + cells in the spleen and blood of chronically infected mice after treatment. (J) Representative dot plots from flow cytometry showing the frequencies of IFN-γ- and TNF-α-producing tetramer + CD8 + T cells. (K) Frequencies of IFN-γ- and TNF-α-expressing tetramer + CD8 + T cells after treatment. Splenocytes were restimulated in vitro for 4 h with PMA and ionomycin in the presence of BFA. (L) Frequencies of proliferating tetramer + CD8 + T cells indicated by Ki67 expression. Results are pooled from two independent experiments. (M) Viral load was determined in the spleen 7 days after treatment started. Results are pooled from three independent experiments. Data shown are means ± standard errors of the means (SEM). Statistics were done by one-way ANOVA with Tukey’s multiple-comparison posttest.

Article Snippet: The following monoclonal antibodies were used. αCD4 (clone RM4-5), αCD8 (clone 53-6.7), αCD80 (clone 16-10A1), αPD-1 (clone J43), and αCD43 (clone 1B11) were obtained from BD Biosciences Pharmingen. αGzmB (clone GB12) and αTOX (clone TXRX10) antibodies were purchased from Invitrogen. αFoxp3 (clone FJK-16s), Ki67 (clone SolA15), and CD127 (clone A7R34) antibodies were purchased from eBioscience. αIFN-y (clone XMG1.2), αTNF-α (clone MP6-XT22), αTCF1 (clone 7F11A10), and CXCR5 (clone L138D7) antibodies were obtained from BioLegend.

Techniques: Infection, Staining, Flow Cytometry, Expressing, In Vivo, Cytotoxicity Assay, In Vitro

Journal: mBio

Article Title: A Combination of Anti-PD-L1 Treatment and Therapeutic Vaccination Facilitates Improved Retroviral Clearance via Reactivation of Highly Exhausted T Cells

doi: 10.1128/mBio.02121-20

Figure Lengend Snippet: Professional APCs experience maturation by combination therapy. Chronically FV-infected mice were treated twice with αPD-L1 or an isotype control (Iso) antibody with or without therapeutic NPV. Splenocytes were analyzed 7 days after the initial treatment. Mean fluorescent intensity (MFI) data are shown for the expression of CD80 by CD11c + F4/80 – DCs (A) and F4/80 + macrophages (B). The results of 2 independent experiments are illustrated. Data represent means ± SEM. Statistics were done by one-way ANOVA with Tukey’s multiple-comparisons posttest.

Article Snippet: The following monoclonal antibodies were used. αCD4 (clone RM4-5), αCD8 (clone 53-6.7), αCD80 (clone 16-10A1), αPD-1 (clone J43), and αCD43 (clone 1B11) were obtained from BD Biosciences Pharmingen. αGzmB (clone GB12) and αTOX (clone TXRX10) antibodies were purchased from Invitrogen. αFoxp3 (clone FJK-16s), Ki67 (clone SolA15), and CD127 (clone A7R34) antibodies were purchased from eBioscience. αIFN-y (clone XMG1.2), αTNF-α (clone MP6-XT22), αTCF1 (clone 7F11A10), and CXCR5 (clone L138D7) antibodies were obtained from BioLegend.

Techniques: Infection, Expressing

Journal: mBio

Article Title: A Combination of Anti-PD-L1 Treatment and Therapeutic Vaccination Facilitates Improved Retroviral Clearance via Reactivation of Highly Exhausted T Cells

doi: 10.1128/mBio.02121-20

Figure Lengend Snippet: Differentiated effector cells are induced by therapeutic vaccination during chronic retrovirus infection, but not by a PD-L1 blockade. Chronically FV-infected mice were treated twice with αPD-L1 or an isotype control (Iso) antibody with or without therapeutic NPV. CD8 + T cell immunity was analyzed 7 days after the initial treatment. (A) Representative flow cytometry dot plots for the identification of SLECs by Klrg1 and CD127 expression. (B) Frequencies of Klrg1 + CD127 − tetramer + CD8 + SLECs are depicted. (C) Frequencies of Klrg1 – CD127 + tetramer + CD8 + cells are shown. The results of 2 independent experiments are illustrated. Data represent means ± SEM. Statistics were done by one-way ANOVA with Tukey’s multiple-comparison posttest. T EX , exhausted CD8 + T cells.

Article Snippet: The following monoclonal antibodies were used. αCD4 (clone RM4-5), αCD8 (clone 53-6.7), αCD80 (clone 16-10A1), αPD-1 (clone J43), and αCD43 (clone 1B11) were obtained from BD Biosciences Pharmingen. αGzmB (clone GB12) and αTOX (clone TXRX10) antibodies were purchased from Invitrogen. αFoxp3 (clone FJK-16s), Ki67 (clone SolA15), and CD127 (clone A7R34) antibodies were purchased from eBioscience. αIFN-y (clone XMG1.2), αTNF-α (clone MP6-XT22), αTCF1 (clone 7F11A10), and CXCR5 (clone L138D7) antibodies were obtained from BioLegend.

Techniques: Infection, Flow Cytometry, Expressing

Journal: mBio

Article Title: A Combination of Anti-PD-L1 Treatment and Therapeutic Vaccination Facilitates Improved Retroviral Clearance via Reactivation of Highly Exhausted T Cells

doi: 10.1128/mBio.02121-20

Figure Lengend Snippet: Enhanced reactivation of differentiated exhausted CD8 + T cells. Chronically FV-infected mice were treated twice with αPD-L1 or an isotype control (Iso) antibody with or without therapeutic NPV. CD8 + T cell immunity was analyzed 7 days after the initial treatment in the spleen. (A) Percentages of PD-1 expression by CD8 + T cells during chronic FV infection. (B) PD-1 expression by tetramer + CD8 + T cells after treatment. (C) Percentages of Ki67-expressing TOX + or TOX – Tet + CD8 + T cells. (D) Frequencies of TOX-expressing cells among PD-1 + tetramer + CD8 + T cells. (E) Representative flow cytometry dot plots illustrate the expression of TOX by the two PD-1 + subpopulations of Tet + CD8 + T cells. Percentages of TOX expression by PD-1 lo and PD-1 hi tetramer + CD8 + T cells were analyzed. (F) Frequencies of GzmB-expressing PD-1 + tetramer + CD8 + T cells were analyzed. (G) GzmB-expressing PD-1 hi tetramer + CD8 + T cells were analyzed. Representative flow cytometry dot plots demonstrate the gating on PD-1 hi -expressing Tet + CD8 + T cells. (H) PD-1 hi -expressing CD8 + T cells were isolated from the spleens of chronically infected mice 7 days after the initial treatment and cultured ex vivo with Gag peptide-loaded and unloaded CFSE-stained target cells. The specific killing of peptide-loaded target cells is depicted. The results from 2 independent experiments were pooled. Bars represent means ± SEM. Statistical analysis was performed by one-way ANOVA with Tukey’s multiple-comparison posttest.

Article Snippet: The following monoclonal antibodies were used. αCD4 (clone RM4-5), αCD8 (clone 53-6.7), αCD80 (clone 16-10A1), αPD-1 (clone J43), and αCD43 (clone 1B11) were obtained from BD Biosciences Pharmingen. αGzmB (clone GB12) and αTOX (clone TXRX10) antibodies were purchased from Invitrogen. αFoxp3 (clone FJK-16s), Ki67 (clone SolA15), and CD127 (clone A7R34) antibodies were purchased from eBioscience. αIFN-y (clone XMG1.2), αTNF-α (clone MP6-XT22), αTCF1 (clone 7F11A10), and CXCR5 (clone L138D7) antibodies were obtained from BioLegend.

Techniques: Infection, Expressing, Flow Cytometry, Isolation, Cell Culture, Ex Vivo, Staining

Journal: mBio

Article Title: A Combination of Anti-PD-L1 Treatment and Therapeutic Vaccination Facilitates Improved Retroviral Clearance via Reactivation of Highly Exhausted T Cells

doi: 10.1128/mBio.02121-20

Figure Lengend Snippet: The timing of therapeutic vaccination is critical for effective combination therapy with a PD-L1 blockade. (A) Chronically FV-infected mice were treated twice with αPD-L1 or an isotype control (Iso) antibody starting at 6 weeks postinfection. Groups of mice received either therapeutic vaccination with CpG- and GagL 85–93 -functionalized CaP nanoparticles alone or in addition to a PD-L1 blockade 4 days after the treatment start. Seven days after therapeutic vaccination, the CD8 + T cell response was analyzed. (B) Viral load was determined in the spleen 7 days after therapeutic vaccination. (C) Percentages of Gag-specific tetramer + CD8 + T cells are shown. (D) Frequencies of IFN-γ- and TNF-α-expressing tetramer + CD8 + T cells after treatment. Splenocytes were restimulated in vitro for 4 h. (E) Frequencies of GzmB-expressing CD43 + tetramer + CD8 + T cells. (F) Frequencies of GzmB-expressing PD-1 hi tetramer + CD8 + T cells. Numbers of CXCR5 + (G) or CXCR5 – (H) tetramer + CD8 + T cells are shown. The results of 2 independent experiments were pooled. Bars represent means ± SEM. Statistical analysis was performed by one-way ANOVA with Tukey’s multiple-comparison posttest.

Article Snippet: The following monoclonal antibodies were used. αCD4 (clone RM4-5), αCD8 (clone 53-6.7), αCD80 (clone 16-10A1), αPD-1 (clone J43), and αCD43 (clone 1B11) were obtained from BD Biosciences Pharmingen. αGzmB (clone GB12) and αTOX (clone TXRX10) antibodies were purchased from Invitrogen. αFoxp3 (clone FJK-16s), Ki67 (clone SolA15), and CD127 (clone A7R34) antibodies were purchased from eBioscience. αIFN-y (clone XMG1.2), αTNF-α (clone MP6-XT22), αTCF1 (clone 7F11A10), and CXCR5 (clone L138D7) antibodies were obtained from BioLegend.

Techniques: Infection, Expressing, In Vitro

Journal: Frontiers in Oncology

Article Title: 4-1BB Agonists: Multi-Potent Potentiators of Tumor Immunity

doi: 10.3389/fonc.2015.00117

Figure Lengend Snippet: Combinations with 4-1BB targeted therapies .

Article Snippet: While both the BMS and Pfizer/Merck trials of α4-1BB/αPD-1 combination therapy seek to translate the impressive pre-clinical efficacy of this combination, it remains to be seen, especially for Urelumab, whether patients will be able to tolerate sufficient doses of these two antibodies to realize this potential.

Techniques: Activity Assay, Expressing, Cell Function Assay, Irradiation

Journal: bioRxiv

Article Title: Stromal HIF2 Regulates Immune Suppression in the Pancreatic Cancer Microenvironment

doi: 10.1101/2021.05.21.445190

Figure Lengend Snippet: (A) UMAP of scRNA-seq analysis of 22,635 cells isolated from KPF CAF-HIF2 WT tumors (10,703 cells; n = 3 mice) and KPF CAF-HIF2 KO tumors (11,932 cells; n = 3 mice). Cell types were identified through graph-based clustering followed by manual annotation using marker genes. (B) Percentage of myeloid cells in each tumor. (C) M2-polarized TAMs were identified within the myeloid cell population via expression of Arg1 and Mrc1. (D) Immunosuppressive TAMs were identified within the myeloid cell population via expression of Cd274 ( Pdl1 ) and Cd86 ( B7-2 ). (E) Violin plots showing findings on scRNA-seq analysis of Ctla4 in KPF CAF-HIF2 WT and KO tumors in all identified cell types. (F) Left: Representative IHC images of CAF-HIF2 WT and KO tumors stained for FoxP3 (n = 5-6/group); scale bars, 50 µm. Right: Quantification of FoxP3+ Tregs per field. (G) Schematic for administration of PT2399 + αCTLA4 in a syngeneic flank KPC model. i.p., intraperitoneal; o.g., oral gavage; b.i.d., bid in die (twice a day). (H) Tumor growth curve from (A) (n = 10/group). Veh, vehicle; P , by Mann–Whitney U test. (I) Schematic for administration of PT2399 + αCTLA4/αPD1 in a syngeneic orthotopic KPC model. (J) Kaplan-Meier curves showing percentage survival for (C) (n = 10/group); P , by log-rank test. All error bars represent mean ± SEM; P , by Student’s t test unless otherwise noted. See also Supplementary Figure 6 and Supplementary Table 2.

Article Snippet: After 2 weeks of recovery, murine αCTLA4 (clone 9D9, Merck) and murine αPD1 (muDX400, Merck) or isotype control were administered IP every 4 days at 20 μg/mouse, 200 μg/mouse, and 220 μg/mouse, respectively for 2 weeks.

Techniques: Isolation, Marker, Expressing, Staining, MANN-WHITNEY

Journal: Biodrugs

Article Title: Targeting Co-Stimulatory Receptors of the TNF Superfamily for Cancer Immunotherapy

doi: 10.1007/s40259-022-00573-3

Figure Lengend Snippet: Tumor necrosis factor super family ( TNF-SF) agonistic monospecific antibodies in clinical studies ( www.clinicaltrials.gov )

Article Snippet: , , Adv. or metastatic malignancies , Ipilimumab (αCTLA-4)/Nivolumab (αPD-1) , I/II , Completed (11/2021) , NCT03126110 , Incyte Corporation.

Techniques: Clinical Proteomics

Journal: Biodrugs

Article Title: Targeting Co-Stimulatory Receptors of the TNF Superfamily for Cancer Immunotherapy

doi: 10.1007/s40259-022-00573-3

Figure Lengend Snippet: Tumor necrosis factor super family ( TNF-SF) agonistic monospecific antibodies in clinical studies ( www.clinicaltrials.gov )

Article Snippet: , , Adv. solid tumors , Durvalumab (αPD-1)/Tremelimumab (αCTLA-4) , I , Completed (08/2019) , NCT02705482 , MedImmune LLC.

Techniques: Clinical Proteomics

Journal: Biodrugs

Article Title: Targeting Co-Stimulatory Receptors of the TNF Superfamily for Cancer Immunotherapy

doi: 10.1007/s40259-022-00573-3

Figure Lengend Snippet: Agonistic tumor necrosis factor super family (TNF-SF) ligand-fusion proteins in clinical studies ( www.clinicaltrials.gov )

Article Snippet: , , Adv. solid tumors , Durvalumab (αPD-1)/Tremelimumab (αCTLA-4) , I , Completed (08/2019) , NCT02705482 , MedImmune LLC.

Techniques: