αpd1 antibody Search Results


αpd-1 j43  (Becton Dickinson)
90
Becton Dickinson αpd-1 j43
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 <t>αPD-L1</t> 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.
αpd 1 J43, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/αpd-1 j43/product/Becton Dickinson
Average 90 stars, based on 1 article reviews
αpd-1 j43 - by Bioz Stars, 2026-03
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murine αpd1 (mudx400  (Merck & Co)
90
Merck & Co murine αpd1 (mudx400
(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 + <t>αCTLA4/αPD1</t> 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.
Murine αpd1 (Mudx400, supplied by Merck & Co, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/murine αpd1 (mudx400/product/Merck & Co
Average 90 stars, based on 1 article reviews
murine αpd1 (mudx400 - by Bioz Stars, 2026-03
90/100 stars
  Buy from Supplier

Image Search Results


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.

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

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.

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

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.

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

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.

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

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.

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

(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.

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