Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Targeting the NAT10-HDAC4 positive feedback loop counteracts immunosuppression in breast cancer

doi: 10.1186/s13046-025-03638-7

Figure Lengend Snippet: Inhibition of NAT10 enhances the efficacy of anti–PD-L1 therapy in vivo . A Schematic diagram of the tumor model and treatment schedule with anti-PD-L1 administration. B–D Representative images ( B ), tumor growth curves ( C ), and tumor weights ( D ) of mouse 4T1 tumors from shNC, shNAT10, anti-PD-L1, or the combination of shNAT10 and anti-PD-L1 ( n = 6). E-F Representative mIF images of mouse tumor tissues showing staining for T cells ( E ), Tregs ( F ), and MDSCs ( F ) (scale bar, 20 μm). G - H ELISA analysis of GzmB (G) and IFN-γ (H) levels in the serum of mice from different groups. I Representative IHC staining of PD-L1, CD8, and GzmB in breast cancer tissues ( n = 220), with statistical analysis of staining intensity (scale bar, 200 μm). J Correlation of PD-L1 and CD8, and PD-L1 and GzmB protein levels in breast cancer tissues ( n = 220) assessed by IHC. All data are presented as mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: In the CD8+ T cell depletion assay, mice were administered anti-Mouse CD8α antibody (Clone 2.43) (MedChemExpress, #HY-P990790, China) intraperitoneally at a dose of 300 μg daily for three consecutive days.

Techniques: Inhibition, In Vivo, Staining, Enzyme-linked Immunosorbent Assay, Immunohistochemistry

Journal: Journal for immunotherapy of cancer

Article Title: Angiotensin receptor blocker attacks armored and cold tumors and boosts immune checkpoint blockade.

doi: 10.1136/jitc-2024-009327

Figure Lengend Snippet: Figure 1 AGTR1 highly expresses in armored and cold tumors and predicts low response to immune checkpoint blockade (ICB) therapy. (A) Heatmap showing the targets expression of concomitant medications in the three immuno-collagenic subtypes in the TCGA-BLCA cohort. (B) Boxplot showing high expression of AGTR1 in armored and cold tumors in the TCGA- BLCA cohort. Horizontal lines in the boxplots represent the median value, and the lower and upper hinges correspond to the first and third quartiles, respectively. Significance was calculated using the ANOVA with Tukey’s multiple-comparison test. ns, non-significance, ***p<0.001. (C) Prognostic value of AGTR1 in the TCGA-BLCA cohort. Subgroups for survival analysis were divided by the best cut-off point. Significance was calculated using the log-rank test. *p<0.05. (D) Representative images uncovering stromal and immune markers in the three immuno-collagenic subtypes in the in-house BLCA cohort. Staining data of HE, Masson, and PD-L1 IHC from our previous study 10 was used as controls. A total of 61 samples were analyzed due to 2 samples losses during multiple staining procedures. (E–G) Expression of PD-L1, CD8, and AGTR1 in the three immuno- collagenic subtypes in the in-house BLCA cohort. Data was presented as mean±SD. Significance was calculated using the Kruskal-Wallis test with the Dunn’s multiple-comparison test for (E). Significance was calculated using the ANOVA with Tukey’s multiple-comparison test for (F and G). ns, non-significance, *p<0.05, ***p<0.001. (H) Prognostic value of AGTR1 in the in- house BLCA cohort. Subgroups for survival analysis were divided by the value of 5%. Significance was calculated using the log-rank test. (I) Predictive value and prognostic value of AGTR1 in the IMvigor210 cohort. Subgroups for survival analysis were divided by the best cut-off point. Significance was calculated using the Student t-test (left) and the log-rank test (right). *p<0.05, ***p<0.001. (J, K) Predictive value of AGTR1 in the GSE135222 and the merged BRCA cohorts. Significance was calculated using the Mann-Whitney test (J) and the Student’s t-test (K). *p<0.05, ***p<0.001. ANOVA, analysis of variance; BLCA, bladder cancer; TCGA, The Cancer Genome Atlas.

Article Snippet: The PD- 1 in vivo mAb (catalog BE0273) was purchased from BioXCell (Lebanon, USA), and the CD8 in vivo mAb (catalog A2102) was purchased from Selleck (Shanghai, China).

Techniques: Expressing, Medications, Comparison, Staining, MANN-WHITNEY

Journal: Journal for immunotherapy of cancer

Article Title: Angiotensin receptor blocker attacks armored and cold tumors and boosts immune checkpoint blockade.

doi: 10.1136/jitc-2024-009327

Figure Lengend Snippet: Figure 5 ARB reverses CAFs-mediated T cell inhibition and exerts immune-dependent tumor suppressive role. (A) The positive rate of AGTR1 in CAFs in 10 BRCA samples in our published cohort. Based on the percentage of AGTR1+ cells in CAFs, BRCA patients were divided into AGTR1-positive and AGTR1-negative groups based on 10%. (B) Boxplot showing exhausted and cytotoxic signature scores in CD8+ T cells in AGTR1-positive and AGTR1-negative groups. Horizontal lines in the boxplots represent the median value, and the lower and upper hinges correspond to the first and third quartiles, respectively. Significance was calculated using the Student’s t-test. ***p<0.001. (C) Schematic protocol of co-culture of CAFs and T cells. (D, E) Differentially expressed genes and expression of activated markers in T cells identified by T cell activation PCR array. (F) Relative content of cytokines in supernatant from the co-culture system was assessed by ELISA assay. Data was presented as mean±SD. Significance was calculated with the Student’s t-test. **p<0.01. (G) Representative images uncovering CD8+ T cell infiltration in samples with low and high AGTR1 expression in the in-house BLCA cohort and quantitative analysis. Significance was calculated using the Pearson test. (H) Tumor growth curve of mice treated with PBS, losartan, and losartan+anti-CD8 antibody. (I) Representative images showing the tumors harvested from mice bearing Lewis cells treated with PBS, losartan, and losartan+anti-CD8 antibody, and weight of the harvested tumors. Data was presented as mean±SD. Significance was calculated with one-way ANOVA with Tukey’s multiple-comparison test. ns, non-significance, *p<0.05. BRCA, breast cancer; ANOVA, analysis of variance; ARB, angiotensin receptor blocker; BLCA, bladder cancer; CAFs, cancer-associated fibroblasts.

Article Snippet: The PD- 1 in vivo mAb (catalog BE0273) was purchased from BioXCell (Lebanon, USA), and the CD8 in vivo mAb (catalog A2102) was purchased from Selleck (Shanghai, China).

Techniques: Inhibition, Co-Culture Assay, Expressing, Activation Assay, Enzyme-linked Immunosorbent Assay, Comparison

Journal: Journal for immunotherapy of cancer

Article Title: Angiotensin receptor blocker attacks armored and cold tumors and boosts immune checkpoint blockade.

doi: 10.1136/jitc-2024-009327

Figure Lengend Snippet: Figure 6 Angiotensin receptor blocker (ARB) shapes an inflamed tumor microenvironment (TME) and enhances immunotherapy in vivo. (A) Tumor growth curve of mice treated with PBS, losartan, anti-PD-1 antibody, and the combination. (B) Representative images showing the tumors harvested from mice bearing Lewis cells treated with PBS, losartan, anti- PD-1 antibody, and the combination, and weight of the harvested tumors. Data was presented as mean±SD. Significance was calculated with one-way ANOVA with Tukey’s multiple-comparison test. ns, non-significance, *p<0.05, ***p<0.001. (C) Representative images showing structure of heart, liver, and kidney from mice in different groups. (D) Representative images showing the levels of collagen, AGTR1, CD8, and Ki67 in tumor tissues from mice in different groups. (E) Representative results of flow cytometry analysis of total CD8+ T cells represented by CD3+CD8+ and quantitative analysis. Data was presented as mean±SD. Significance was calculated with one-way ANOVA with Tukey’s multiple-comparison test. ns, non-significance, **p<0.01, ***p<0.001. (F) Representative results of flow cytometry analysis of myeloid-derived suppressor cells (MDSC) represented by CD11b+Gr-1+ and quantitative analysis. Data was presented as mean±SD. Significance was calculated with one- way ANOVA with Tukey’s multiple-comparison test. ns, non-significance, ***p<0.001. PBS: phosphate buffer saline; ANOVA, analysis of variance.

Article Snippet: The PD- 1 in vivo mAb (catalog BE0273) was purchased from BioXCell (Lebanon, USA), and the CD8 in vivo mAb (catalog A2102) was purchased from Selleck (Shanghai, China).

Techniques: In Vivo, Comparison, Flow Cytometry, Derivative Assay, Saline

Journal: Immunity

Article Title: The white adipose tissue is a reservoir for memory T cells that promotes protective memory responses to infection

doi: 10.1016/j.immuni.2017.11.009

Figure Lengend Snippet: DATA AND SOFTWARE AVAILABILITY

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Anti-mouse α4β7 (DATK32) eBioscience Cat# 12-5887-82 Anti-mouse B220 (RA3-6B2) BD Biosciences Cat# 562290 Anti-mouse CCR2(FAB5538P) R&D Systems Cat# FAB5538P Anti-mouse CCR7 (4B12) Biolegend Cat# 120107 Anti-mouse CCR9 (CW1.2) eBioscience Cat# 12-1991-82 Anti-mouse CD3 (17A2) eBioscience Cat# 47-0032-82 Anti-mouse CD4 (RM4-5) Biolegend Cat# 100548 Anti-mouse CD8β (H35–17.2) eBioscience Cat# 25-0083-82 Anti-mouse CD11a (M17/4) eBioscience Cat# 12-0111-82 Anti-mouse CD11b (M1/70) BD Biosciences Cat# 563015 Anti-mouse CD11c (N418) Biolegend Cat# 117336 Anti-mouse CD44 (IM7) eBioscience Cat# 25-0441-82 Anti-mouse CD45 (30-F11) eBioscience Cat# 47-0451-82 Anti-mouse CD45.1 (A20) Biolegend Cat# 110737 Anti-mouse CD45.2 (104) Biolegend Cat# 109837 Anti-mouse CD62L (MEL-14) eBioscience Cat# 11-0621-85 Anti-mouse CD69 (H1.2F3) eBioscience Cat# 12-0691-83 Anti-mouse CD90.2 (30-H12) Biolegend Cat# 105331 Anti-mouse CD103 (2E7) eBioscience Cat# 46-1031-82 Anti-mouse CD122 (TMb1) eBioscience Cat# 46-1222-82 Anti-mouse CD127 (A7R34) Biolegend Cat# 135025 Anti-mouse Foxp3 (FJK-16S) eBioscience Cat# 17-5773-82 Anti-mouse γδTCR (GL3) BD Biosciences Cat# 563532 Anti-mouse GATA-3 (L50-823) BD Biosciences Cat# 563349 Anti-mouse IFN-γ (XMG1.2) eBioscience Cat# 48-7311-82 Anti-mouse IL-5 (TRFK5) Fisher Scientific Cat# 554395 Anti-mouse IL-13 (13A) eBioscience Cat# 53-7133-82 Anti-mouse IL-17A (17B7) Biolegend Cat# 506910 Anti-mouse Ki67 (SolA15) eBioscience Cat# 48-5698-82 Anti-mouse KLRG1 (2F1) eBioscience Cat# 12-5893-82 Anti-mouse Ly-6C (HK1.5) Biolegend Cat# 128035 Anti-mouse Ly-6G (IA8) BD Biosciences Cat# 562700 Anti-mouse MHC-II (M5/114.15.2) eBioscience Cat# 56-5321-82 Anti-mouse NK1.1 (PK136) BD Biosciences Cat# 562864 Anti-mouse Siglec F (E50-2440) BD Biosciences Cat# 562757 Anti-mouse T-bet (4B10) Biolegend Cat# 644816 Anti-mouse TCRβ (H57–597) eBioscience Cat# 47-5961-82 Anti-mouse TNF-α (MP6-XT22) BD Biosciences Cat# 558000 Anti-human CD3 (SP34-2) BD Biosciences Cat# 558124 Anti-human CD4 (OKT4) eBioscience Cat# 93-0048-42 Anti-human CD8 (SK1) eBioscience Cat# 46-0087-42 Anti-human CD28 (CD28.2) Beckman Coulter Cat# 6607111 Anti-human CD69 (FN50) BD Biosciences Cat# 557745 Anti-human CD95 (DX2) Biolegend Cat# 305610 Anti-human CD103 (HML-1) Beckman Coulter Cat# IM 1856 Anti-human IFN-γ (B27) BD Biosciences Cat# 561024 Anti-human IL-13 (JES10-5A2) BD Biosciences Cat# 554571 Anti-human TNF-α (MAb11) eBiosciences Cat# 25-7349-82 Anti-mouse CD4 (GK1.1) (depletion) BioXcell Cat# BE0003-1 Anti-mouse CD8α (2.43) (depletion) BioXcell Cat# BE0061 Bacterial and Virus Strains Yersinia pseudotuberculosis (32777) Laboratory of I.E.B.

Techniques: Software, Recombinant, Enzyme-linked Immunosorbent Assay, Isolation, SYBR Green Assay, Staining, Microarray

Journal: Frontiers in Immunology

Article Title: LAMP1 targeting of the large T antigen of Merkel cell polyomavirus results in potent CD4 T cell responses and tumor inhibition

doi: 10.3389/fimmu.2023.1253568

Figure Lengend Snippet: ITI-3000 induces spatial immune T cell infiltration into tumors. Experimental design shown in (A) . C57BL/6 mice were given 5E4 B16-LT tumor cells subcutaneously in the right flank. Starting on day 3 post tumor injection, mice were vaccinated twice, weekly, with 40µg of ITI-3000 or control vector via intradermal injection followed by electroporation. Tumors were collected and formalin fixed on day 15 post tumor injection, followed by a sucrose gradient, paraffin embedding, and sectioning. The cellular composition of the tumor microenvironment on FFPE slides was determined by multiplex-IHC including percent of CD3 + (green), CD8 + (magenta) (B) , and CD4 + (red) (C) cells. One representative section (displayed both as whole tumor section (right) and magnified area of interest (left)) of each group is shown in (D) The total cell population was determined by nuclear DAPI staining (blue). Sections were analyzed by AI gating in the Biodock platform. N=5. Statistical significance was calculated by unpaired Student’s t-test. The "*" symbol represents a p-value of <0.05.

Article Snippet: A murine 5-color panel including CD3, CD4, CD8, FoxP3, and DAPI was done at Ultivue Inc. (Cambridge, MA).

Techniques: Injection, Control, Plasmid Preparation, Electroporation, Multiplex Assay, Staining