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pe cyanine7 anti mouse cd4  (Elabscience Biotechnology)


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    Elabscience Biotechnology pe cyanine7 anti mouse cd4
    IL22 upregulates PD-L1 expression in gastric cancer cells through the UPR IRE1α–XBP1 axis. A, mIF images show the alterations of PD-L1 + tumor cells (purple), <t>CD4</t> + (green), and CD8 + (red) T cells in orthotopic gastric cancer tumors from control and Abhd16a -knockdown mice following IL22 treatment. Scale bar, 50 μm. B, KEGG pathway enrichment analysis of RNA-seq data of gastric cancer tissues with or without IL22 treatment. C, RT-PCR was used to assess the mRNA expression of key downstream molecules of the UPR branches ( XBP1 , ATF4 , ATF6 ) in control and IL22RA1 -knockdown gastric cancer cells. D, Western blotting analysis of PD-L1 and XBP1s levels in control and IL22RA1 -knockdown MGC-803 cells treated with IL22 (100 μg/L). E, Western blotting detection of PD-L1 and XBP1s levels in XBP1- knockdown MGC-803 cells treated with IL22 and MGC-803 cells treated with IL22 or XBP1s inhibitor (STF083010, 30 μmol/L) in combination with IL22. F, The binding sequence of XBP1 on the CD274 promoter. G and H, ChIP ( G ) and luciferase reporter assay ( H ) showing the transcriptional regulation of CD274 by XBP1s under IL22 stimulation. I, Orthotopic gastric cancer mouse models ( n = 5 per group) were injected with anti-IL22 (200 μg per mouse), anti-CD90.2 antibody (150 μg per mouse), anti-CD90.2 antibody in combination with IL22 (500 ng per mouse), or anti-CD90.2 antibody in combination with XBP1s inhibitors (STF083010, 30 mg/kg) and IL22 for 2 weeks. IHC analysis was used to show IL22, XBP1s, and PD-L1 levels in gastric cancer tissues. Scale bar, 200 μm. J, Tumor volume of orthotopic gastric cancer models under treatments the same as in I . *, P < 0.05; **, P < 0.01; ***, P < 0.001; ns, nonsignificant.
    Pe Cyanine7 Anti Mouse Cd4, supplied by Elabscience Biotechnology, used in various techniques. Bioz Stars score: 94/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/pe+cyanine7+anti+mouse+cd4/PE%2FCyanine7+Anti-Mouse+CD4+Antibody/pmc13080324-162-48-51
    Average 94 stars, based on 8 article reviews
    pe cyanine7 anti mouse cd4 - by Bioz Stars, 2026-09
    94/100 stars

    Images

    1) Product Images from "Nerves Stimulate Cross-talk Between Gastric Cancer and Group 3 Innate Lymphoid Cells to Enhance Immunosuppression"

    Article Title: Nerves Stimulate Cross-talk Between Gastric Cancer and Group 3 Innate Lymphoid Cells to Enhance Immunosuppression

    Journal: Cancer Research

    doi: 10.1158/0008-5472.CAN-25-3092

    IL22 upregulates PD-L1 expression in gastric cancer cells through the UPR IRE1α–XBP1 axis. A, mIF images show the alterations of PD-L1 + tumor cells (purple), CD4 + (green), and CD8 + (red) T cells in orthotopic gastric cancer tumors from control and Abhd16a -knockdown mice following IL22 treatment. Scale bar, 50 μm. B, KEGG pathway enrichment analysis of RNA-seq data of gastric cancer tissues with or without IL22 treatment. C, RT-PCR was used to assess the mRNA expression of key downstream molecules of the UPR branches ( XBP1 , ATF4 , ATF6 ) in control and IL22RA1 -knockdown gastric cancer cells. D, Western blotting analysis of PD-L1 and XBP1s levels in control and IL22RA1 -knockdown MGC-803 cells treated with IL22 (100 μg/L). E, Western blotting detection of PD-L1 and XBP1s levels in XBP1- knockdown MGC-803 cells treated with IL22 and MGC-803 cells treated with IL22 or XBP1s inhibitor (STF083010, 30 μmol/L) in combination with IL22. F, The binding sequence of XBP1 on the CD274 promoter. G and H, ChIP ( G ) and luciferase reporter assay ( H ) showing the transcriptional regulation of CD274 by XBP1s under IL22 stimulation. I, Orthotopic gastric cancer mouse models ( n = 5 per group) were injected with anti-IL22 (200 μg per mouse), anti-CD90.2 antibody (150 μg per mouse), anti-CD90.2 antibody in combination with IL22 (500 ng per mouse), or anti-CD90.2 antibody in combination with XBP1s inhibitors (STF083010, 30 mg/kg) and IL22 for 2 weeks. IHC analysis was used to show IL22, XBP1s, and PD-L1 levels in gastric cancer tissues. Scale bar, 200 μm. J, Tumor volume of orthotopic gastric cancer models under treatments the same as in I . *, P < 0.05; **, P < 0.01; ***, P < 0.001; ns, nonsignificant.
    Figure Legend Snippet: IL22 upregulates PD-L1 expression in gastric cancer cells through the UPR IRE1α–XBP1 axis. A, mIF images show the alterations of PD-L1 + tumor cells (purple), CD4 + (green), and CD8 + (red) T cells in orthotopic gastric cancer tumors from control and Abhd16a -knockdown mice following IL22 treatment. Scale bar, 50 μm. B, KEGG pathway enrichment analysis of RNA-seq data of gastric cancer tissues with or without IL22 treatment. C, RT-PCR was used to assess the mRNA expression of key downstream molecules of the UPR branches ( XBP1 , ATF4 , ATF6 ) in control and IL22RA1 -knockdown gastric cancer cells. D, Western blotting analysis of PD-L1 and XBP1s levels in control and IL22RA1 -knockdown MGC-803 cells treated with IL22 (100 μg/L). E, Western blotting detection of PD-L1 and XBP1s levels in XBP1- knockdown MGC-803 cells treated with IL22 and MGC-803 cells treated with IL22 or XBP1s inhibitor (STF083010, 30 μmol/L) in combination with IL22. F, The binding sequence of XBP1 on the CD274 promoter. G and H, ChIP ( G ) and luciferase reporter assay ( H ) showing the transcriptional regulation of CD274 by XBP1s under IL22 stimulation. I, Orthotopic gastric cancer mouse models ( n = 5 per group) were injected with anti-IL22 (200 μg per mouse), anti-CD90.2 antibody (150 μg per mouse), anti-CD90.2 antibody in combination with IL22 (500 ng per mouse), or anti-CD90.2 antibody in combination with XBP1s inhibitors (STF083010, 30 mg/kg) and IL22 for 2 weeks. IHC analysis was used to show IL22, XBP1s, and PD-L1 levels in gastric cancer tissues. Scale bar, 200 μm. J, Tumor volume of orthotopic gastric cancer models under treatments the same as in I . *, P < 0.05; **, P < 0.01; ***, P < 0.001; ns, nonsignificant.

    Techniques Used: Expressing, Control, Knockdown, RNA Sequencing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Binding Assay, Sequencing, Luciferase, Reporter Assay, Injection

    Combination therapy enhances the anti–PD-L1 immunotherapeutic effect in gastric cancer. A and B, After tumor formation, the orthotopic gastric cancer mice ( n = 5 per group) were treated with anti–PD-L1 (100 μg per mouse), GPR34 inhibitor (20 mg/kg), or XBP1s inhibitor (30 mg/kg) every 3 days or ACh inhibitor (2.5 mg/kg) daily. Combinations of anti–PD-L1 with each inhibitor followed the every 3-day dosing schedule for a total duration of 2 weeks via i.p. injection. Living images were used to monitor tumor progression at 5-day intervals from the time of drug administration ( A ); IHC and mIF were performed to detect PD-L1 and XBP1s levels and proportions of CD4 + (green) and CD8 + (red) T cells in gastric cancer tissues at the end of treatments ( B ). Scale bars, 1.000e+5 –∼ 5.000e + 5 p/s/cm 2 /sr for living images; 200 μm for IHC; 50 μm for immunofluorescence. C and D, Representative images ( C ) and tumor volume ( D ) of subcutaneous tumors. The administration protocol for the mice was consistent with the description provided in A and B . **, P < 0.01; ***, P < 0.001.
    Figure Legend Snippet: Combination therapy enhances the anti–PD-L1 immunotherapeutic effect in gastric cancer. A and B, After tumor formation, the orthotopic gastric cancer mice ( n = 5 per group) were treated with anti–PD-L1 (100 μg per mouse), GPR34 inhibitor (20 mg/kg), or XBP1s inhibitor (30 mg/kg) every 3 days or ACh inhibitor (2.5 mg/kg) daily. Combinations of anti–PD-L1 with each inhibitor followed the every 3-day dosing schedule for a total duration of 2 weeks via i.p. injection. Living images were used to monitor tumor progression at 5-day intervals from the time of drug administration ( A ); IHC and mIF were performed to detect PD-L1 and XBP1s levels and proportions of CD4 + (green) and CD8 + (red) T cells in gastric cancer tissues at the end of treatments ( B ). Scale bars, 1.000e+5 –∼ 5.000e + 5 p/s/cm 2 /sr for living images; 200 μm for IHC; 50 μm for immunofluorescence. C and D, Representative images ( C ) and tumor volume ( D ) of subcutaneous tumors. The administration protocol for the mice was consistent with the description provided in A and B . **, P < 0.01; ***, P < 0.001.

    Techniques Used: Injection, Immunofluorescence

    Related Articles

    Flow Cytometry:

    Article Title: Nerves Stimulates Crosstalk between Gastric Cancer and Group 3 Innate Lymphoid Cells to Enhance Immunosuppression
    Article Snippet: 2 The immunosuppressive tumor microenvironment (TME) enables cancer cells to 2 evade clinical immunotherapies.. Neural networks are vital components of the TME, 3 and interactions between cancer cells, neuronal cells, and immune cells mediate 4 immunosuppression.. Hence, understanding the mechanisms of intercellular crosstalk 5 could inform immunomodulatory approaches to enhance immunotherapy efficacy.

    Article Title: Nerves Stimulate Cross-talk Between Gastric Cancer and Group 3 Innate Lymphoid Cells to Enhance Immunosuppression
    Article Snippet: .. The antibodies used for flow cytometry: Brilliant Violet 605 anti-mouse CD127 (BioLegend, cat. #135025, RRID: AB_2562114, 5 μL/1 × 10 6 cells), FITC anti-mouse CD3 (BioLegend, cat. #100203, RRID: AB_312660, 2 μL/1 × 10 6 cells), APC anti-mouse CD3 (Elabscience, cat. #E-AB-F1013E, RRID: AB_3675272, 5 μL/1×10 6 cells), PE/Cyanine7 anti-mouse CD4 (Elabscience, cat. #E-AB-F1097H, 5 μL/1 × 10 6 cells), FITC Anti-Mouse CD8a (Elabscience, cat. #E-AB-F1104UC, 5 μL/1 × 10 6 cells), FITC anti-mouse CD19 (BioLegend, cat. #152403, RRID: AB_2629812, 0.25 μL/1 × 10 6 cells), FITC anti-mouse CD11c (BioLegend, cat. #117305, RRID: AB_313774, 0.5 μL/1 × 10 6 cells), FITC anti-mouse NK1.1 (BioLegend, cat. #108705, RRID: AB_313392, 0.5 μL/1 × 10 6 cells), Brilliant Violet 421 anti-mouse CD45 (BioLegend, cat. #103133, RRID: AB_10899570, 1 μL/1 × 10 6 cells), PE anti-mouse RORγt (BD Biosciences, cat. #562607, RRID: AB_11153137, 2 μL/1 × 10 6 cells), PerCP/Cyanine5.5 anti-mouse IL22 (BioLegend, cat. #516411, RRID: AB_2563373, 5 μL/1 × 10 6 cells), AF647 anti-STAT3 phospho (BioLegend, cat. #651007, RRID: AB_2572085, 5 μL/1 × 10 6 cells), PE anti-mouse CD45 (BioLegend, cat. #157604, RRID: AB_2876536, 1.25 μL/1 × 10 6 cells), APC anti-mouse CD8b (BioLegend, cat. #126613, RRID: AB_2562774, 0.625 μL/1 × 10 6 cells), APC anti-mouse CD4 (BioLegend, cat. #100411, RRID: AB_312696, 1.25 μL/1 × 10 6 cells), APC anti-mouse CD206 (BioLegend, cat. #141707, RRID: AB_10896057, 2.5 μL/1 × 10 6 cells), FITC anti-mouse F4/80 (BioLegend, cat. #157309, RRID: AB_2876535, 2 μL/1 × 10 6 cells), FITC anti-mouse CD25 (BioLegend, cat. #101907, RRID: AB_961210, 2 μL/1 × 10 6 cells), AF700 anti-mouse FOXP3 (BioLegend, cat. #126421, RRID: AB_2750492, 0.12 μL/1 × 10 6 cells), PE anti-mouse Ly6G (BioLegend, cat. #127607, RRID: AB_1186104, 1.25 μL/1 × 10 6 cells), APC anti-mouse CD274 (Elabscience, cat. #E-AB-F1132E, 5 μL/1 × 10 6 cells), PerCP-Cyanine5.5 anti–T-bet (eBioscience, cat. #45-5825-80, RRID: AB_953658, 0.25 μg/1 × 10 6 cells), PE/Dazzle 594 anti-mouse CD273 (BioLegend, cat. #107215, RRID: AB_2728124, 0.25 μg/1 × 10 6 cells), Brilliant Violet 421 anti-mouse CD274 (BioLegend, cat. #124315, RRID: AB_10897097, 5 μL/1 × 10 6 cells), and PE anti-mouse MHC-I (H-2Kk; BioLegend, cat. #114907, RRID: AB_313614, 0.25 μg/1 × 10 6 cells). .. FITC anti-human CD3 (Thermo Fisher Scientific, cat. #11-0038-42, RRID: AB_2043831, 5 μL/1 × 10 6 cells), PE anti-human CD127 (BioLegend, cat. #351304, RRID: AB_10720185, 5 μL/1 × 10 6 cells), Brilliant Violet 421 anti-human CD294 (BioLegend, cat. #350112, RRID: AB_2562468, 5 μL/1 × 10 6 cells), APC anti-human CD117 (BioLegend, cat. #313206, RRID: AB_314985, 5 μL/1 × 10 6 cells), FITC anti-human CD19 (eBioscience, cat. #11-0199-42, RRID: AB_10669461, 5 μL/1 × 10 6 cells), FITC anti-human CD14 (eBioscience, cat. #11-0149-42, RRID: AB_10597597, 5 μL/1 × 10 6 cells), BV650 anti-human CD45 (eBioscience, cat. #416-0459-42, RRID: AB_2925684, 5 μL/1 × 10 6 cells), iFluor 647 anti-Ki67 (HUABIO, cat. #HA720163F, RRID: AB_3072100, 1 μL/1 × 10 6 cells), PE/Cyanine7 anti-human CD274 (Elabscience, cat. #E-AB-F1133H, 5 μL/1 × 10 6 cells), and PE anti-human CD273 (Elabscience, cat. #E-AB-F1175D, 5 μL/1 × 10 6 cells).

    Immunopeptidomics:

    Article Title: Nerves Stimulates Crosstalk between Gastric Cancer and Group 3 Innate Lymphoid Cells to Enhance Immunosuppression
    Article Snippet: 2 The immunosuppressive tumor microenvironment (TME) enables cancer cells to 2 evade clinical immunotherapies.. Neural networks are vital components of the TME, 3 and interactions between cancer cells, neuronal cells, and immune cells mediate 4 immunosuppression.. Hence, understanding the mechanisms of intercellular crosstalk 5 could inform immunomodulatory approaches to enhance immunotherapy efficacy.

    Article Title: Nerves Stimulate Cross-talk Between Gastric Cancer and Group 3 Innate Lymphoid Cells to Enhance Immunosuppression
    Article Snippet: .. The antibodies used for flow cytometry: Brilliant Violet 605 anti-mouse CD127 (BioLegend, cat. #135025, RRID: AB_2562114, 5 μL/1 × 10 6 cells), FITC anti-mouse CD3 (BioLegend, cat. #100203, RRID: AB_312660, 2 μL/1 × 10 6 cells), APC anti-mouse CD3 (Elabscience, cat. #E-AB-F1013E, RRID: AB_3675272, 5 μL/1×10 6 cells), PE/Cyanine7 anti-mouse CD4 (Elabscience, cat. #E-AB-F1097H, 5 μL/1 × 10 6 cells), FITC Anti-Mouse CD8a (Elabscience, cat. #E-AB-F1104UC, 5 μL/1 × 10 6 cells), FITC anti-mouse CD19 (BioLegend, cat. #152403, RRID: AB_2629812, 0.25 μL/1 × 10 6 cells), FITC anti-mouse CD11c (BioLegend, cat. #117305, RRID: AB_313774, 0.5 μL/1 × 10 6 cells), FITC anti-mouse NK1.1 (BioLegend, cat. #108705, RRID: AB_313392, 0.5 μL/1 × 10 6 cells), Brilliant Violet 421 anti-mouse CD45 (BioLegend, cat. #103133, RRID: AB_10899570, 1 μL/1 × 10 6 cells), PE anti-mouse RORγt (BD Biosciences, cat. #562607, RRID: AB_11153137, 2 μL/1 × 10 6 cells), PerCP/Cyanine5.5 anti-mouse IL22 (BioLegend, cat. #516411, RRID: AB_2563373, 5 μL/1 × 10 6 cells), AF647 anti-STAT3 phospho (BioLegend, cat. #651007, RRID: AB_2572085, 5 μL/1 × 10 6 cells), PE anti-mouse CD45 (BioLegend, cat. #157604, RRID: AB_2876536, 1.25 μL/1 × 10 6 cells), APC anti-mouse CD8b (BioLegend, cat. #126613, RRID: AB_2562774, 0.625 μL/1 × 10 6 cells), APC anti-mouse CD4 (BioLegend, cat. #100411, RRID: AB_312696, 1.25 μL/1 × 10 6 cells), APC anti-mouse CD206 (BioLegend, cat. #141707, RRID: AB_10896057, 2.5 μL/1 × 10 6 cells), FITC anti-mouse F4/80 (BioLegend, cat. #157309, RRID: AB_2876535, 2 μL/1 × 10 6 cells), FITC anti-mouse CD25 (BioLegend, cat. #101907, RRID: AB_961210, 2 μL/1 × 10 6 cells), AF700 anti-mouse FOXP3 (BioLegend, cat. #126421, RRID: AB_2750492, 0.12 μL/1 × 10 6 cells), PE anti-mouse Ly6G (BioLegend, cat. #127607, RRID: AB_1186104, 1.25 μL/1 × 10 6 cells), APC anti-mouse CD274 (Elabscience, cat. #E-AB-F1132E, 5 μL/1 × 10 6 cells), PerCP-Cyanine5.5 anti–T-bet (eBioscience, cat. #45-5825-80, RRID: AB_953658, 0.25 μg/1 × 10 6 cells), PE/Dazzle 594 anti-mouse CD273 (BioLegend, cat. #107215, RRID: AB_2728124, 0.25 μg/1 × 10 6 cells), Brilliant Violet 421 anti-mouse CD274 (BioLegend, cat. #124315, RRID: AB_10897097, 5 μL/1 × 10 6 cells), and PE anti-mouse MHC-I (H-2Kk; BioLegend, cat. #114907, RRID: AB_313614, 0.25 μg/1 × 10 6 cells). .. FITC anti-human CD3 (Thermo Fisher Scientific, cat. #11-0038-42, RRID: AB_2043831, 5 μL/1 × 10 6 cells), PE anti-human CD127 (BioLegend, cat. #351304, RRID: AB_10720185, 5 μL/1 × 10 6 cells), Brilliant Violet 421 anti-human CD294 (BioLegend, cat. #350112, RRID: AB_2562468, 5 μL/1 × 10 6 cells), APC anti-human CD117 (BioLegend, cat. #313206, RRID: AB_314985, 5 μL/1 × 10 6 cells), FITC anti-human CD19 (eBioscience, cat. #11-0199-42, RRID: AB_10669461, 5 μL/1 × 10 6 cells), FITC anti-human CD14 (eBioscience, cat. #11-0149-42, RRID: AB_10597597, 5 μL/1 × 10 6 cells), BV650 anti-human CD45 (eBioscience, cat. #416-0459-42, RRID: AB_2925684, 5 μL/1 × 10 6 cells), iFluor 647 anti-Ki67 (HUABIO, cat. #HA720163F, RRID: AB_3072100, 1 μL/1 × 10 6 cells), PE/Cyanine7 anti-human CD274 (Elabscience, cat. #E-AB-F1133H, 5 μL/1 × 10 6 cells), and PE anti-human CD273 (Elabscience, cat. #E-AB-F1175D, 5 μL/1 × 10 6 cells).



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    Effect of KVBCPs on the T lymphocyte differentiation of splenic lymphocytes of Balb/c mice. (A) Representative flow cytometry plots of CD3 + <t>CD4</t> + T cells after treatment with KVBCPs (100 μg/mL) for 48 h; (B) The change of the proportions of CD4 + after treatment with KVBCPs (1, 10, 100 μg/mL) for 48 h. Data are expressed as the mean ± SD (n = 3). ## P < 0.01 vs control group; ∗∗ P < 0.01 vs Con A group, one way ANOVA.
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    CXCL13 is highly expressed in T cells from responders to ICB therapy. a). UMAP plot displaying 237231 CD3 + T cells across 8 cancer types: BCC (basal cell carcinoma, n = 11 patients; n = 22 samples), SCC (squamous cell carcinoma, n = 3 patients; n = 8 samples), BLCA (bladder cancer, n = 6 patients; n = 6 samples), TNBC (triple‐negative breast cancer, n = 8 patients; n = 14 samples), RCC (Renal cell carcinoma, n = 7 patients; n = 7 samples), HNSCC (head and neck cancer, n = 4 patients; n = 8 samples), NSCLC (non‐small‐cell lung cancer, n = 36 patients; n = 47 samples), and PRAD (prostate cancer, n = 10 patients; n = 12 samples), categorized into 13 distinct cell types. b). Comparison of differential genes in CD3 + , CD8 + , and <t>CD4</t> + T cells in the response and non‐response groups. c,d). UMAP plots (c) and dot plots (d) showing the expression of CXCL13 in CD3 + , CD8 + , and CD4 + T cells of pre‐treatment non‐response, pre‐treatment response, post‐treatment non‐response, and post‐treatment response groups. e). CXCL13 expression levels in responders and non‐responders across multiple tumor cohorts treated with ICB: HNSC, melanoma cohort 1 (anti‐PD‐1 and anti‐CTLA‐4), melanoma cohort 2 (anti‐PD‐1), STAD (stomach adenocarcinoma), and BLCA. f). Comparison of CXCL13 expression in pre‐treatment and post‐treatment groups from melanoma cohort 1 and melanoma cohort 2 treated with ICB. g). Kaplan‐Meier survival curves stratified by high and low CXCL13 expression in ICB‐treated patients from BLCA, melanoma cohort 1, and melanoma cohort 2. h). Heatmap showing the correlation of CXCL13 expression with various cell types across tumor types in the TCGA dataset. Cell‐type deconvolution was performed using TIMER2.0, with significant correlations ( p < 0.05) marked by solid indicators.
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    CXCL13 is highly expressed in T cells from responders to ICB therapy. a). UMAP plot displaying 237231 CD3 + T cells across 8 cancer types: BCC (basal cell carcinoma, n = 11 patients; n = 22 samples), SCC (squamous cell carcinoma, n = 3 patients; n = 8 samples), BLCA (bladder cancer, n = 6 patients; n = 6 samples), TNBC (triple‐negative breast cancer, n = 8 patients; n = 14 samples), RCC (Renal cell carcinoma, n = 7 patients; n = 7 samples), HNSCC (head and neck cancer, n = 4 patients; n = 8 samples), NSCLC (non‐small‐cell lung cancer, n = 36 patients; n = 47 samples), and PRAD (prostate cancer, n = 10 patients; n = 12 samples), categorized into 13 distinct cell types. b). Comparison of differential genes in CD3 + , CD8 + , and <t>CD4</t> + T cells in the response and non‐response groups. c,d). UMAP plots (c) and dot plots (d) showing the expression of CXCL13 in CD3 + , CD8 + , and CD4 + T cells of pre‐treatment non‐response, pre‐treatment response, post‐treatment non‐response, and post‐treatment response groups. e). CXCL13 expression levels in responders and non‐responders across multiple tumor cohorts treated with ICB: HNSC, melanoma cohort 1 (anti‐PD‐1 and anti‐CTLA‐4), melanoma cohort 2 (anti‐PD‐1), STAD (stomach adenocarcinoma), and BLCA. f). Comparison of CXCL13 expression in pre‐treatment and post‐treatment groups from melanoma cohort 1 and melanoma cohort 2 treated with ICB. g). Kaplan‐Meier survival curves stratified by high and low CXCL13 expression in ICB‐treated patients from BLCA, melanoma cohort 1, and melanoma cohort 2. h). Heatmap showing the correlation of CXCL13 expression with various cell types across tumor types in the TCGA dataset. Cell‐type deconvolution was performed using TIMER2.0, with significant correlations ( p < 0.05) marked by solid indicators.
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    Image Search Results


    IL22 upregulates PD-L1 expression in gastric cancer cells through the UPR IRE1α–XBP1 axis. A, mIF images show the alterations of PD-L1 + tumor cells (purple), CD4 + (green), and CD8 + (red) T cells in orthotopic gastric cancer tumors from control and Abhd16a -knockdown mice following IL22 treatment. Scale bar, 50 μm. B, KEGG pathway enrichment analysis of RNA-seq data of gastric cancer tissues with or without IL22 treatment. C, RT-PCR was used to assess the mRNA expression of key downstream molecules of the UPR branches ( XBP1 , ATF4 , ATF6 ) in control and IL22RA1 -knockdown gastric cancer cells. D, Western blotting analysis of PD-L1 and XBP1s levels in control and IL22RA1 -knockdown MGC-803 cells treated with IL22 (100 μg/L). E, Western blotting detection of PD-L1 and XBP1s levels in XBP1- knockdown MGC-803 cells treated with IL22 and MGC-803 cells treated with IL22 or XBP1s inhibitor (STF083010, 30 μmol/L) in combination with IL22. F, The binding sequence of XBP1 on the CD274 promoter. G and H, ChIP ( G ) and luciferase reporter assay ( H ) showing the transcriptional regulation of CD274 by XBP1s under IL22 stimulation. I, Orthotopic gastric cancer mouse models ( n = 5 per group) were injected with anti-IL22 (200 μg per mouse), anti-CD90.2 antibody (150 μg per mouse), anti-CD90.2 antibody in combination with IL22 (500 ng per mouse), or anti-CD90.2 antibody in combination with XBP1s inhibitors (STF083010, 30 mg/kg) and IL22 for 2 weeks. IHC analysis was used to show IL22, XBP1s, and PD-L1 levels in gastric cancer tissues. Scale bar, 200 μm. J, Tumor volume of orthotopic gastric cancer models under treatments the same as in I . *, P < 0.05; **, P < 0.01; ***, P < 0.001; ns, nonsignificant.

    Journal: Cancer Research

    Article Title: Nerves Stimulate Cross-talk Between Gastric Cancer and Group 3 Innate Lymphoid Cells to Enhance Immunosuppression

    doi: 10.1158/0008-5472.CAN-25-3092

    Figure Lengend Snippet: IL22 upregulates PD-L1 expression in gastric cancer cells through the UPR IRE1α–XBP1 axis. A, mIF images show the alterations of PD-L1 + tumor cells (purple), CD4 + (green), and CD8 + (red) T cells in orthotopic gastric cancer tumors from control and Abhd16a -knockdown mice following IL22 treatment. Scale bar, 50 μm. B, KEGG pathway enrichment analysis of RNA-seq data of gastric cancer tissues with or without IL22 treatment. C, RT-PCR was used to assess the mRNA expression of key downstream molecules of the UPR branches ( XBP1 , ATF4 , ATF6 ) in control and IL22RA1 -knockdown gastric cancer cells. D, Western blotting analysis of PD-L1 and XBP1s levels in control and IL22RA1 -knockdown MGC-803 cells treated with IL22 (100 μg/L). E, Western blotting detection of PD-L1 and XBP1s levels in XBP1- knockdown MGC-803 cells treated with IL22 and MGC-803 cells treated with IL22 or XBP1s inhibitor (STF083010, 30 μmol/L) in combination with IL22. F, The binding sequence of XBP1 on the CD274 promoter. G and H, ChIP ( G ) and luciferase reporter assay ( H ) showing the transcriptional regulation of CD274 by XBP1s under IL22 stimulation. I, Orthotopic gastric cancer mouse models ( n = 5 per group) were injected with anti-IL22 (200 μg per mouse), anti-CD90.2 antibody (150 μg per mouse), anti-CD90.2 antibody in combination with IL22 (500 ng per mouse), or anti-CD90.2 antibody in combination with XBP1s inhibitors (STF083010, 30 mg/kg) and IL22 for 2 weeks. IHC analysis was used to show IL22, XBP1s, and PD-L1 levels in gastric cancer tissues. Scale bar, 200 μm. J, Tumor volume of orthotopic gastric cancer models under treatments the same as in I . *, P < 0.05; **, P < 0.01; ***, P < 0.001; ns, nonsignificant.

    Article Snippet: The antibodies used for flow cytometry: Brilliant Violet 605 anti-mouse CD127 (BioLegend, cat. #135025, RRID: AB_2562114, 5 μL/1 × 10 6 cells), FITC anti-mouse CD3 (BioLegend, cat. #100203, RRID: AB_312660, 2 μL/1 × 10 6 cells), APC anti-mouse CD3 (Elabscience, cat. #E-AB-F1013E, RRID: AB_3675272, 5 μL/1×10 6 cells), PE/Cyanine7 anti-mouse CD4 (Elabscience, cat. #E-AB-F1097H, 5 μL/1 × 10 6 cells), FITC Anti-Mouse CD8a (Elabscience, cat. #E-AB-F1104UC, 5 μL/1 × 10 6 cells), FITC anti-mouse CD19 (BioLegend, cat. #152403, RRID: AB_2629812, 0.25 μL/1 × 10 6 cells), FITC anti-mouse CD11c (BioLegend, cat. #117305, RRID: AB_313774, 0.5 μL/1 × 10 6 cells), FITC anti-mouse NK1.1 (BioLegend, cat. #108705, RRID: AB_313392, 0.5 μL/1 × 10 6 cells), Brilliant Violet 421 anti-mouse CD45 (BioLegend, cat. #103133, RRID: AB_10899570, 1 μL/1 × 10 6 cells), PE anti-mouse RORγt (BD Biosciences, cat. #562607, RRID: AB_11153137, 2 μL/1 × 10 6 cells), PerCP/Cyanine5.5 anti-mouse IL22 (BioLegend, cat. #516411, RRID: AB_2563373, 5 μL/1 × 10 6 cells), AF647 anti-STAT3 phospho (BioLegend, cat. #651007, RRID: AB_2572085, 5 μL/1 × 10 6 cells), PE anti-mouse CD45 (BioLegend, cat. #157604, RRID: AB_2876536, 1.25 μL/1 × 10 6 cells), APC anti-mouse CD8b (BioLegend, cat. #126613, RRID: AB_2562774, 0.625 μL/1 × 10 6 cells), APC anti-mouse CD4 (BioLegend, cat. #100411, RRID: AB_312696, 1.25 μL/1 × 10 6 cells), APC anti-mouse CD206 (BioLegend, cat. #141707, RRID: AB_10896057, 2.5 μL/1 × 10 6 cells), FITC anti-mouse F4/80 (BioLegend, cat. #157309, RRID: AB_2876535, 2 μL/1 × 10 6 cells), FITC anti-mouse CD25 (BioLegend, cat. #101907, RRID: AB_961210, 2 μL/1 × 10 6 cells), AF700 anti-mouse FOXP3 (BioLegend, cat. #126421, RRID: AB_2750492, 0.12 μL/1 × 10 6 cells), PE anti-mouse Ly6G (BioLegend, cat. #127607, RRID: AB_1186104, 1.25 μL/1 × 10 6 cells), APC anti-mouse CD274 (Elabscience, cat. #E-AB-F1132E, 5 μL/1 × 10 6 cells), PerCP-Cyanine5.5 anti–T-bet (eBioscience, cat. #45-5825-80, RRID: AB_953658, 0.25 μg/1 × 10 6 cells), PE/Dazzle 594 anti-mouse CD273 (BioLegend, cat. #107215, RRID: AB_2728124, 0.25 μg/1 × 10 6 cells), Brilliant Violet 421 anti-mouse CD274 (BioLegend, cat. #124315, RRID: AB_10897097, 5 μL/1 × 10 6 cells), and PE anti-mouse MHC-I (H-2Kk; BioLegend, cat. #114907, RRID: AB_313614, 0.25 μg/1 × 10 6 cells).

    Techniques: Expressing, Control, Knockdown, RNA Sequencing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Binding Assay, Sequencing, Luciferase, Reporter Assay, Injection

    Combination therapy enhances the anti–PD-L1 immunotherapeutic effect in gastric cancer. A and B, After tumor formation, the orthotopic gastric cancer mice ( n = 5 per group) were treated with anti–PD-L1 (100 μg per mouse), GPR34 inhibitor (20 mg/kg), or XBP1s inhibitor (30 mg/kg) every 3 days or ACh inhibitor (2.5 mg/kg) daily. Combinations of anti–PD-L1 with each inhibitor followed the every 3-day dosing schedule for a total duration of 2 weeks via i.p. injection. Living images were used to monitor tumor progression at 5-day intervals from the time of drug administration ( A ); IHC and mIF were performed to detect PD-L1 and XBP1s levels and proportions of CD4 + (green) and CD8 + (red) T cells in gastric cancer tissues at the end of treatments ( B ). Scale bars, 1.000e+5 –∼ 5.000e + 5 p/s/cm 2 /sr for living images; 200 μm for IHC; 50 μm for immunofluorescence. C and D, Representative images ( C ) and tumor volume ( D ) of subcutaneous tumors. The administration protocol for the mice was consistent with the description provided in A and B . **, P < 0.01; ***, P < 0.001.

    Journal: Cancer Research

    Article Title: Nerves Stimulate Cross-talk Between Gastric Cancer and Group 3 Innate Lymphoid Cells to Enhance Immunosuppression

    doi: 10.1158/0008-5472.CAN-25-3092

    Figure Lengend Snippet: Combination therapy enhances the anti–PD-L1 immunotherapeutic effect in gastric cancer. A and B, After tumor formation, the orthotopic gastric cancer mice ( n = 5 per group) were treated with anti–PD-L1 (100 μg per mouse), GPR34 inhibitor (20 mg/kg), or XBP1s inhibitor (30 mg/kg) every 3 days or ACh inhibitor (2.5 mg/kg) daily. Combinations of anti–PD-L1 with each inhibitor followed the every 3-day dosing schedule for a total duration of 2 weeks via i.p. injection. Living images were used to monitor tumor progression at 5-day intervals from the time of drug administration ( A ); IHC and mIF were performed to detect PD-L1 and XBP1s levels and proportions of CD4 + (green) and CD8 + (red) T cells in gastric cancer tissues at the end of treatments ( B ). Scale bars, 1.000e+5 –∼ 5.000e + 5 p/s/cm 2 /sr for living images; 200 μm for IHC; 50 μm for immunofluorescence. C and D, Representative images ( C ) and tumor volume ( D ) of subcutaneous tumors. The administration protocol for the mice was consistent with the description provided in A and B . **, P < 0.01; ***, P < 0.001.

    Article Snippet: The antibodies used for flow cytometry: Brilliant Violet 605 anti-mouse CD127 (BioLegend, cat. #135025, RRID: AB_2562114, 5 μL/1 × 10 6 cells), FITC anti-mouse CD3 (BioLegend, cat. #100203, RRID: AB_312660, 2 μL/1 × 10 6 cells), APC anti-mouse CD3 (Elabscience, cat. #E-AB-F1013E, RRID: AB_3675272, 5 μL/1×10 6 cells), PE/Cyanine7 anti-mouse CD4 (Elabscience, cat. #E-AB-F1097H, 5 μL/1 × 10 6 cells), FITC Anti-Mouse CD8a (Elabscience, cat. #E-AB-F1104UC, 5 μL/1 × 10 6 cells), FITC anti-mouse CD19 (BioLegend, cat. #152403, RRID: AB_2629812, 0.25 μL/1 × 10 6 cells), FITC anti-mouse CD11c (BioLegend, cat. #117305, RRID: AB_313774, 0.5 μL/1 × 10 6 cells), FITC anti-mouse NK1.1 (BioLegend, cat. #108705, RRID: AB_313392, 0.5 μL/1 × 10 6 cells), Brilliant Violet 421 anti-mouse CD45 (BioLegend, cat. #103133, RRID: AB_10899570, 1 μL/1 × 10 6 cells), PE anti-mouse RORγt (BD Biosciences, cat. #562607, RRID: AB_11153137, 2 μL/1 × 10 6 cells), PerCP/Cyanine5.5 anti-mouse IL22 (BioLegend, cat. #516411, RRID: AB_2563373, 5 μL/1 × 10 6 cells), AF647 anti-STAT3 phospho (BioLegend, cat. #651007, RRID: AB_2572085, 5 μL/1 × 10 6 cells), PE anti-mouse CD45 (BioLegend, cat. #157604, RRID: AB_2876536, 1.25 μL/1 × 10 6 cells), APC anti-mouse CD8b (BioLegend, cat. #126613, RRID: AB_2562774, 0.625 μL/1 × 10 6 cells), APC anti-mouse CD4 (BioLegend, cat. #100411, RRID: AB_312696, 1.25 μL/1 × 10 6 cells), APC anti-mouse CD206 (BioLegend, cat. #141707, RRID: AB_10896057, 2.5 μL/1 × 10 6 cells), FITC anti-mouse F4/80 (BioLegend, cat. #157309, RRID: AB_2876535, 2 μL/1 × 10 6 cells), FITC anti-mouse CD25 (BioLegend, cat. #101907, RRID: AB_961210, 2 μL/1 × 10 6 cells), AF700 anti-mouse FOXP3 (BioLegend, cat. #126421, RRID: AB_2750492, 0.12 μL/1 × 10 6 cells), PE anti-mouse Ly6G (BioLegend, cat. #127607, RRID: AB_1186104, 1.25 μL/1 × 10 6 cells), APC anti-mouse CD274 (Elabscience, cat. #E-AB-F1132E, 5 μL/1 × 10 6 cells), PerCP-Cyanine5.5 anti–T-bet (eBioscience, cat. #45-5825-80, RRID: AB_953658, 0.25 μg/1 × 10 6 cells), PE/Dazzle 594 anti-mouse CD273 (BioLegend, cat. #107215, RRID: AB_2728124, 0.25 μg/1 × 10 6 cells), Brilliant Violet 421 anti-mouse CD274 (BioLegend, cat. #124315, RRID: AB_10897097, 5 μL/1 × 10 6 cells), and PE anti-mouse MHC-I (H-2Kk; BioLegend, cat. #114907, RRID: AB_313614, 0.25 μg/1 × 10 6 cells).

    Techniques: Injection, Immunofluorescence

    Effect of KVBCPs on the T lymphocyte differentiation of splenic lymphocytes of Balb/c mice. (A) Representative flow cytometry plots of CD3 + CD4 + T cells after treatment with KVBCPs (100 μg/mL) for 48 h; (B) The change of the proportions of CD4 + after treatment with KVBCPs (1, 10, 100 μg/mL) for 48 h. Data are expressed as the mean ± SD (n = 3). ## P < 0.01 vs control group; ∗∗ P < 0.01 vs Con A group, one way ANOVA.

    Journal: Journal of Traditional and Complementary Medicine

    Article Title: Polysaccharides derived from alkali-extracted vinegar-baked Radix Bupleuri suppress hyperimmune T lymphocytes and ameliorate skin graft rejection

    doi: 10.1016/j.jtcme.2024.11.006

    Figure Lengend Snippet: Effect of KVBCPs on the T lymphocyte differentiation of splenic lymphocytes of Balb/c mice. (A) Representative flow cytometry plots of CD3 + CD4 + T cells after treatment with KVBCPs (100 μg/mL) for 48 h; (B) The change of the proportions of CD4 + after treatment with KVBCPs (1, 10, 100 μg/mL) for 48 h. Data are expressed as the mean ± SD (n = 3). ## P < 0.01 vs control group; ∗∗ P < 0.01 vs Con A group, one way ANOVA.

    Article Snippet: Foxp3/Transcription Factor Staining Buffer Kit (TNB-0607-KIT), Flow Staining Buffer (1X) (TNB-4222-L500), APC Anti-Mouse CD3 (Lot: C0032032122203), PE-Cyanine7 Anti-Mouse CD4 (Lot: C0041121820603), FITC Anti-Mouse CD25 (Lot: C0251020422353) and PE Anti-Mouse Foxp3(Lot: C2107092418503) were supplied by Tonbo Biosciences (California, USA).

    Techniques: Flow Cytometry, Control

    The effect of KVBCP3 and KVBCP4 on CD3 + and CD4 + /CD8 + infiltration of mice with skin allograft rejection. (A) CD3 + , CD4 + and CD8 + staining of skin graft; (B) The change of CD3 + infiltration after treatment with KVBCPs (n = 3). (C) The change of CD4 + /CD8 + infiltration after treatment with KVBCPs (n = 3). Data are expressed as the mean ± SD. ## P < 0.01 vs Sham control group; ∗ P < 0.05, ∗∗ P < 0.01 vs Model group, one way ANOVA.

    Journal: Journal of Traditional and Complementary Medicine

    Article Title: Polysaccharides derived from alkali-extracted vinegar-baked Radix Bupleuri suppress hyperimmune T lymphocytes and ameliorate skin graft rejection

    doi: 10.1016/j.jtcme.2024.11.006

    Figure Lengend Snippet: The effect of KVBCP3 and KVBCP4 on CD3 + and CD4 + /CD8 + infiltration of mice with skin allograft rejection. (A) CD3 + , CD4 + and CD8 + staining of skin graft; (B) The change of CD3 + infiltration after treatment with KVBCPs (n = 3). (C) The change of CD4 + /CD8 + infiltration after treatment with KVBCPs (n = 3). Data are expressed as the mean ± SD. ## P < 0.01 vs Sham control group; ∗ P < 0.05, ∗∗ P < 0.01 vs Model group, one way ANOVA.

    Article Snippet: Foxp3/Transcription Factor Staining Buffer Kit (TNB-0607-KIT), Flow Staining Buffer (1X) (TNB-4222-L500), APC Anti-Mouse CD3 (Lot: C0032032122203), PE-Cyanine7 Anti-Mouse CD4 (Lot: C0041121820603), FITC Anti-Mouse CD25 (Lot: C0251020422353) and PE Anti-Mouse Foxp3(Lot: C2107092418503) were supplied by Tonbo Biosciences (California, USA).

    Techniques: Staining, Control

    The effect of KVBCP3 and KVBCP4 on immune cytokines and Treg cells of mice with skin allograft rejection. (A) The secretion of IL-2 in serum was determined by Elisa assay (n = 6). (B) The secretion of IL-4 in serum was determined by Elisa assay (n = 6). (C) Representative flow cytometry plots of CD4 + CD25 + Foxp3 + Treg cells after treatment with KVBCPs at the 14th day; (D) The change of the proportions of CD4 + CD25 + Foxp3 + Treg cells after treatment with KVBCPs at the 14th day (n = 3). (E) Representative flow cytometry plots of CD4 + IL-17A + cells after treatment with KVBCPs at the 14th day; (F) The change of the proportions of CD4 + IL-17A + cells after treatment with KVBCPs at the 14th day (n = 3). ∗ P < 0.05, ∗∗ P < 0.01 vs Model group, one way ANOVA.

    Journal: Journal of Traditional and Complementary Medicine

    Article Title: Polysaccharides derived from alkali-extracted vinegar-baked Radix Bupleuri suppress hyperimmune T lymphocytes and ameliorate skin graft rejection

    doi: 10.1016/j.jtcme.2024.11.006

    Figure Lengend Snippet: The effect of KVBCP3 and KVBCP4 on immune cytokines and Treg cells of mice with skin allograft rejection. (A) The secretion of IL-2 in serum was determined by Elisa assay (n = 6). (B) The secretion of IL-4 in serum was determined by Elisa assay (n = 6). (C) Representative flow cytometry plots of CD4 + CD25 + Foxp3 + Treg cells after treatment with KVBCPs at the 14th day; (D) The change of the proportions of CD4 + CD25 + Foxp3 + Treg cells after treatment with KVBCPs at the 14th day (n = 3). (E) Representative flow cytometry plots of CD4 + IL-17A + cells after treatment with KVBCPs at the 14th day; (F) The change of the proportions of CD4 + IL-17A + cells after treatment with KVBCPs at the 14th day (n = 3). ∗ P < 0.05, ∗∗ P < 0.01 vs Model group, one way ANOVA.

    Article Snippet: Foxp3/Transcription Factor Staining Buffer Kit (TNB-0607-KIT), Flow Staining Buffer (1X) (TNB-4222-L500), APC Anti-Mouse CD3 (Lot: C0032032122203), PE-Cyanine7 Anti-Mouse CD4 (Lot: C0041121820603), FITC Anti-Mouse CD25 (Lot: C0251020422353) and PE Anti-Mouse Foxp3(Lot: C2107092418503) were supplied by Tonbo Biosciences (California, USA).

    Techniques: Enzyme-linked Immunosorbent Assay, Flow Cytometry

    CXCL13 is highly expressed in T cells from responders to ICB therapy. a). UMAP plot displaying 237231 CD3 + T cells across 8 cancer types: BCC (basal cell carcinoma, n = 11 patients; n = 22 samples), SCC (squamous cell carcinoma, n = 3 patients; n = 8 samples), BLCA (bladder cancer, n = 6 patients; n = 6 samples), TNBC (triple‐negative breast cancer, n = 8 patients; n = 14 samples), RCC (Renal cell carcinoma, n = 7 patients; n = 7 samples), HNSCC (head and neck cancer, n = 4 patients; n = 8 samples), NSCLC (non‐small‐cell lung cancer, n = 36 patients; n = 47 samples), and PRAD (prostate cancer, n = 10 patients; n = 12 samples), categorized into 13 distinct cell types. b). Comparison of differential genes in CD3 + , CD8 + , and CD4 + T cells in the response and non‐response groups. c,d). UMAP plots (c) and dot plots (d) showing the expression of CXCL13 in CD3 + , CD8 + , and CD4 + T cells of pre‐treatment non‐response, pre‐treatment response, post‐treatment non‐response, and post‐treatment response groups. e). CXCL13 expression levels in responders and non‐responders across multiple tumor cohorts treated with ICB: HNSC, melanoma cohort 1 (anti‐PD‐1 and anti‐CTLA‐4), melanoma cohort 2 (anti‐PD‐1), STAD (stomach adenocarcinoma), and BLCA. f). Comparison of CXCL13 expression in pre‐treatment and post‐treatment groups from melanoma cohort 1 and melanoma cohort 2 treated with ICB. g). Kaplan‐Meier survival curves stratified by high and low CXCL13 expression in ICB‐treated patients from BLCA, melanoma cohort 1, and melanoma cohort 2. h). Heatmap showing the correlation of CXCL13 expression with various cell types across tumor types in the TCGA dataset. Cell‐type deconvolution was performed using TIMER2.0, with significant correlations ( p < 0.05) marked by solid indicators.

    Journal: Advanced Science

    Article Title: CXCL13 Expression Promotes CAR T Cell Antitumor Activity and Potentiates Response to PD‐1 Blockade

    doi: 10.1002/advs.202508095

    Figure Lengend Snippet: CXCL13 is highly expressed in T cells from responders to ICB therapy. a). UMAP plot displaying 237231 CD3 + T cells across 8 cancer types: BCC (basal cell carcinoma, n = 11 patients; n = 22 samples), SCC (squamous cell carcinoma, n = 3 patients; n = 8 samples), BLCA (bladder cancer, n = 6 patients; n = 6 samples), TNBC (triple‐negative breast cancer, n = 8 patients; n = 14 samples), RCC (Renal cell carcinoma, n = 7 patients; n = 7 samples), HNSCC (head and neck cancer, n = 4 patients; n = 8 samples), NSCLC (non‐small‐cell lung cancer, n = 36 patients; n = 47 samples), and PRAD (prostate cancer, n = 10 patients; n = 12 samples), categorized into 13 distinct cell types. b). Comparison of differential genes in CD3 + , CD8 + , and CD4 + T cells in the response and non‐response groups. c,d). UMAP plots (c) and dot plots (d) showing the expression of CXCL13 in CD3 + , CD8 + , and CD4 + T cells of pre‐treatment non‐response, pre‐treatment response, post‐treatment non‐response, and post‐treatment response groups. e). CXCL13 expression levels in responders and non‐responders across multiple tumor cohorts treated with ICB: HNSC, melanoma cohort 1 (anti‐PD‐1 and anti‐CTLA‐4), melanoma cohort 2 (anti‐PD‐1), STAD (stomach adenocarcinoma), and BLCA. f). Comparison of CXCL13 expression in pre‐treatment and post‐treatment groups from melanoma cohort 1 and melanoma cohort 2 treated with ICB. g). Kaplan‐Meier survival curves stratified by high and low CXCL13 expression in ICB‐treated patients from BLCA, melanoma cohort 1, and melanoma cohort 2. h). Heatmap showing the correlation of CXCL13 expression with various cell types across tumor types in the TCGA dataset. Cell‐type deconvolution was performed using TIMER2.0, with significant correlations ( p < 0.05) marked by solid indicators.

    Article Snippet: The following antibodies were used for staining: anti‐mouse CD45.1, BV450 (E‐AB‐F1184UQ), Elabscience, Cat # AF19855; anti‐mouse CD45.1, PE (A20), Biolegend, Cat # 110 708; anti‐mouse CD3ε, PE/Cyanine7 (145‐2C11), Biolegend, Cat # 100 320; anti‐mouse PD‐1, PE/Cyanine7 (RMP1‐30), Biolegend, Cat # 109 110; anti‐mouse CD4, APC/Cy7 (E‐AB‐F1353UJ), Elabscience, Cat # AF18028; anti‐mouse CD4, PerCP/Cyanine5.5 (RM4‐5), Biolegend, Cat # 100 540; anti‐mouse CD4, PE/Cyanine7 (RM4‐5), Biolegend, Cat # 100 527; anti‐mouse CD4, Brilliant Violet 785 (RM4‐5), Biolegend, Cat # 100 552; anti‐mouse CD8a, PE (53‐6.7), Biolegend, Cat # 100 708; anti‐mouse CD8a, APC (53‐6.7), Biolegend, Cat # 100 712; anti‐mouse CD8a, PE/Cyanine7 (53‐6.7), Biolegend, Cat # 100 721; anti‐mouse CD45, PerCP/Cyanine5.5 (30‐F11), Biolegend, Cat # 103 132; anti‐mouse Ly108, PE (330‐AJ), Biolegend, Cat # 134 605; anti‐mouse LAG‐3, PerCP/Cyanine5.5 (C9B7W), Biolegend, Cat # 125 211; anti‐mouse LAG‐3, APC (C9B7W), Biolegend, Cat # 125 210; anti‐mouse CD69, Brilliant Violet 605 (H1.2F3), Biolegend, Cat # 104 530; anti‐mouse/human CD44, Brilliant Violet 510 (IM7), Biolegend, Cat # 103 044; anti‐mouse CD62L, PerCP/Cyanine5.5 (MEL‐14), Biolegend, Cat # 104 432; anti‐mouse CTLA‐4, APC (UC10‐4B9), Biolegend, Cat # 106 309; anti‐human CD3, APC (OKT3), Biolegend, Cat # 317 318; anti‐human CD4, PerCP/Cyanine5.5 (RPA‐T4), Biolegend, Cat # 300 530; anti‐human CD8, APC/Cyanine7 (SK1), Biolegend, Cat # 344 713; anti‐human CD279(PD‐1), PE (A17188A), Biolegend, Cat # 379 209; anti‐human CD366(Tim3), PE/CY7 (F38‐2E2), Biolegend, Cat # 345 014.

    Techniques: Comparison, Expressing

    CXCL13 CAR T cells display enhanced antitumor characteristics. a). GSEA analysis of the mitochondrial membrane potential gene set in bulk RNA‐seq comparing CXCL13 CAR T cells with control CAR T cells. b–d). Flow cytometry analysis of mitochondrial membrane potential (TMRM) in control CAR T and CXCL13 CAR T cells. Representative TMRM peak plots are shown for GFP + (b) and GFP − (c) T cells cultured in vitro on day 10 and for cells isolated from the spleen (d). TMRM: Tetramethylrhodamine, methyl ester. Mean ± SEM, n = 3 (b, c); n = 5 (control CAR T), n = 4 (CXCL13 CAR T) (d). e). Intracellular ATP levels in control CAR T cells and CXCL13 CAR T cells. Mean ± SEM, n = 4. f,g). GSEA analysis of positive regulation of T cell migration gene set in CXCL13 CAR T cells versus control CAR T cells using bulk RNA‐seq data (f) and CXCL13 + cells versus CXCL13 − cells using scRNA‐seq data (g). h). Schematic overview of the transwell migration assay. i,j). The migration ability of CAR T cells was assessed by the transwell assay, showing the proportion (i) and number (j) of cells that migrated through the membrane. Mean ± SEM, n = 4. k–m). C57BL/6 mice injected s.c. with 5 × 10 5 B16‐CD19 cells were treated with 2×10⁶ CAR T cells on day 5 post‐tumor establishment. The number of CAR T cells was detected after 7 days of adoptive transfer in the spleen (k), draining lymph nodes (l), and tumor sites (m). Mean ± SEM, n = 3. n). Percentage of CD4 + T cells among CD3 + T cells from patients with GPC3‐positive solid tumors treated with CAR T cells (non‐responders [NR], n = 6; responders [R], n = 15). Mean ± SEM. o,p). Flow cytometry plots (o) and line graphs (p) showing the percentage of CD4⁺ CAR T cells in vitro from day 4 to day 10. Mean ± SEM, n = 4. q–s). Proportions of CD4 + and CD8 + CAR T cells in blood (q), spleen (r), and draining lymph nodes (s). Mean ± SEM, blood ( n = 6), spleen ( n = 6 for control CAR T; n = 4 for CXCL13 CAR T), draining lymph nodes ( n = 6 for control CAR T; n = 4 for CXCL13 CAR T). * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; two‐tailed unpaired t‐test (b–e,i–n,p–s).

    Journal: Advanced Science

    Article Title: CXCL13 Expression Promotes CAR T Cell Antitumor Activity and Potentiates Response to PD‐1 Blockade

    doi: 10.1002/advs.202508095

    Figure Lengend Snippet: CXCL13 CAR T cells display enhanced antitumor characteristics. a). GSEA analysis of the mitochondrial membrane potential gene set in bulk RNA‐seq comparing CXCL13 CAR T cells with control CAR T cells. b–d). Flow cytometry analysis of mitochondrial membrane potential (TMRM) in control CAR T and CXCL13 CAR T cells. Representative TMRM peak plots are shown for GFP + (b) and GFP − (c) T cells cultured in vitro on day 10 and for cells isolated from the spleen (d). TMRM: Tetramethylrhodamine, methyl ester. Mean ± SEM, n = 3 (b, c); n = 5 (control CAR T), n = 4 (CXCL13 CAR T) (d). e). Intracellular ATP levels in control CAR T cells and CXCL13 CAR T cells. Mean ± SEM, n = 4. f,g). GSEA analysis of positive regulation of T cell migration gene set in CXCL13 CAR T cells versus control CAR T cells using bulk RNA‐seq data (f) and CXCL13 + cells versus CXCL13 − cells using scRNA‐seq data (g). h). Schematic overview of the transwell migration assay. i,j). The migration ability of CAR T cells was assessed by the transwell assay, showing the proportion (i) and number (j) of cells that migrated through the membrane. Mean ± SEM, n = 4. k–m). C57BL/6 mice injected s.c. with 5 × 10 5 B16‐CD19 cells were treated with 2×10⁶ CAR T cells on day 5 post‐tumor establishment. The number of CAR T cells was detected after 7 days of adoptive transfer in the spleen (k), draining lymph nodes (l), and tumor sites (m). Mean ± SEM, n = 3. n). Percentage of CD4 + T cells among CD3 + T cells from patients with GPC3‐positive solid tumors treated with CAR T cells (non‐responders [NR], n = 6; responders [R], n = 15). Mean ± SEM. o,p). Flow cytometry plots (o) and line graphs (p) showing the percentage of CD4⁺ CAR T cells in vitro from day 4 to day 10. Mean ± SEM, n = 4. q–s). Proportions of CD4 + and CD8 + CAR T cells in blood (q), spleen (r), and draining lymph nodes (s). Mean ± SEM, blood ( n = 6), spleen ( n = 6 for control CAR T; n = 4 for CXCL13 CAR T), draining lymph nodes ( n = 6 for control CAR T; n = 4 for CXCL13 CAR T). * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; two‐tailed unpaired t‐test (b–e,i–n,p–s).

    Article Snippet: The following antibodies were used for staining: anti‐mouse CD45.1, BV450 (E‐AB‐F1184UQ), Elabscience, Cat # AF19855; anti‐mouse CD45.1, PE (A20), Biolegend, Cat # 110 708; anti‐mouse CD3ε, PE/Cyanine7 (145‐2C11), Biolegend, Cat # 100 320; anti‐mouse PD‐1, PE/Cyanine7 (RMP1‐30), Biolegend, Cat # 109 110; anti‐mouse CD4, APC/Cy7 (E‐AB‐F1353UJ), Elabscience, Cat # AF18028; anti‐mouse CD4, PerCP/Cyanine5.5 (RM4‐5), Biolegend, Cat # 100 540; anti‐mouse CD4, PE/Cyanine7 (RM4‐5), Biolegend, Cat # 100 527; anti‐mouse CD4, Brilliant Violet 785 (RM4‐5), Biolegend, Cat # 100 552; anti‐mouse CD8a, PE (53‐6.7), Biolegend, Cat # 100 708; anti‐mouse CD8a, APC (53‐6.7), Biolegend, Cat # 100 712; anti‐mouse CD8a, PE/Cyanine7 (53‐6.7), Biolegend, Cat # 100 721; anti‐mouse CD45, PerCP/Cyanine5.5 (30‐F11), Biolegend, Cat # 103 132; anti‐mouse Ly108, PE (330‐AJ), Biolegend, Cat # 134 605; anti‐mouse LAG‐3, PerCP/Cyanine5.5 (C9B7W), Biolegend, Cat # 125 211; anti‐mouse LAG‐3, APC (C9B7W), Biolegend, Cat # 125 210; anti‐mouse CD69, Brilliant Violet 605 (H1.2F3), Biolegend, Cat # 104 530; anti‐mouse/human CD44, Brilliant Violet 510 (IM7), Biolegend, Cat # 103 044; anti‐mouse CD62L, PerCP/Cyanine5.5 (MEL‐14), Biolegend, Cat # 104 432; anti‐mouse CTLA‐4, APC (UC10‐4B9), Biolegend, Cat # 106 309; anti‐human CD3, APC (OKT3), Biolegend, Cat # 317 318; anti‐human CD4, PerCP/Cyanine5.5 (RPA‐T4), Biolegend, Cat # 300 530; anti‐human CD8, APC/Cyanine7 (SK1), Biolegend, Cat # 344 713; anti‐human CD279(PD‐1), PE (A17188A), Biolegend, Cat # 379 209; anti‐human CD366(Tim3), PE/CY7 (F38‐2E2), Biolegend, Cat # 345 014.

    Techniques: Membrane, RNA Sequencing, Control, Flow Cytometry, Cell Culture, In Vitro, Isolation, Migration, Transwell Migration Assay, Transwell Assay, Injection, Adoptive Transfer Assay, Two Tailed Test