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anti cd4 pe  (Miltenyi Biotec)


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    Structured Review

    Miltenyi Biotec anti cd4 pe
    Anti Cd4 Pe, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 95/100, based on 123 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/cd4+pe/CD4+Antibody%2C+anti-mouse%2C+REAfinity/pmc13355016-7-0-2
    Average 95 stars, based on 123 article reviews
    anti cd4 pe - by Bioz Stars, 2026-11
    95/100 stars

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    Related Articles

    Bioprocessing:

    Article Title: Ex vivo enhancement of CD8+ T cell activity using functionalized hydrogel encapsulating tonsil-derived lymphatic endothelial cells
    Article Snippet: .. The following anti-human monoclonal antibodies were used in this study: 1) CD44 PE, CD73 APC, CD90 FITC, CD105PerCP-Cy5.5, and CD11b/CD19/CD34/CD45/HLA-DR PE (Human MSC Analysis Kit #562245, BD Biosciences) for TMSCs; 2) podoplanin APC, VEGF-R3 PE (both Biolegend), ALEXA FLUOR ® 488-conjugated Prox-1 (Bioss, Woburn, MA., USA), and ALEXA FLUOR ® 488-conjugated LYVE-1 (R&D Systems, Minneapolis, MN., USA) for T-LECs; 3) CD4 PE, CD8 APC, CD44 FITC, CD62L PE, CD69PE (Miltenyi Biotec, Westphalia, Germany), IL-2 PE, and IFN-γ FITC(Biolegend) for T cells sorting or STCs; 4) PD-L1 PE (Biolegend) for TME. ..

    Article Title: Longitudinal Assessment of Multiple Immunological and Inflammatory Parameters during Successful DAA Therapy in HCV Monoinfected and HIV/HCV Coinfected Subjects
    Article Snippet: Lysing solution (BD Biosciences Pharmagen, Milano, Italy) was added to the samples, and after additional 10 min of incubation, the samples were analyzed using a MACS Quant flow cytometer (Miltenyi Biotec, Bergisch Gladbach, Cologne, Germany). .. The anti-human monoclonal antibodies used were CD45-VioBlue, HLA-DR-FITC, CD38-APC, CD8-PerCP and CD4-PE (all from Miltenyi Biotec, Bergisch Gladbach, Germany). ..

    Article Title: Targeting the melanoma-associated antigen CSPG4 with HLA-C*07:01-restricted T-cell receptors.
    Article Snippet: .. The following monoclonal antibodies were used: CD4-PE (clone 13B8.2; Beckman Coulter, Brea, CA, USA), CD8-APC (clone B9.11; Beckman Coulter), CD8-PE (clone B9.11; Beckman Coulter), antimurine TCR-FITC (clone H57-697; Biolegend, San Diego, CA, USA), anti-human TCR-APC (clone IP26, Biolegend) and CSPG4-PE (REA1041, Milteny). .. For surface staining of MHC-I, tumor cells were stained by incubation with supernatant of the hybridomaW6/32 (kindly provided by Dr. P. Parham, Departments of Structural Biology and Microbiology and Immunology, Stanford University, Stanford, CA, USA) followed by staining with polyclonal goat-anti-mouse-FITC (Biozol, Eching, Germany).

    Staining:

    Article Title: From DGCR8 expression analysis to diseased pathways in 22q11.2 deletion syndrome
    Article Snippet: Thawed PBMCs of patients and healthy controls, obtained as described above, were treated with IFN2alpha (300 U/mL, Miltenyi Biotech) for 48 and 72 hours at 37°C, 5% CO2. .. Cells were surface stained with CD3 VioBlue (Miltenyi), CD4 PE (Miltenyi) and CD45 V500-C (BD). .. Cells were then fixed and permeabilized with Fix&Perm Biolegend kit and stained with anti-STAT1 AF647 (BD) or corresponding isotype control AF647 (Biolegend).

    Article Title: Therapeutic activity of retroviral replicating vector-mediated gene therapy in combination with anti-PD-1 antibody in a murine pancreatic cancer model.
    Article Snippet: Toca 511, a tumor-selective retroviral replicating vector encoding the yeast cytosine deaminase (yCD) gene, exerts direct antitumor effects through intratumoral prodrug 5-fluorocytosine (5-FC) conversion to active drug 5-fluorouracil by yCD, and has demonstrated therapeutic efficacy in preclinical and clinical trials of various cancers.. Toca 511/5-FC treatment may also induce antitumor immunity.. Here, we first examined antitumor immune responses activated by Toca 511/5-FC treatment in an immunocompetent murine pancreatic cancer model. We then evaluated the therapeutic effects achieved in combination with anti-programmed cell death protein 1 antibody.

    Flow Cytometry:

    Article Title: Therapeutic activity of retroviral replicating vector-mediated gene therapy in combination with anti-PD-1 antibody in a murine pancreatic cancer model.
    Article Snippet: Toca 511, a tumor-selective retroviral replicating vector encoding the yeast cytosine deaminase (yCD) gene, exerts direct antitumor effects through intratumoral prodrug 5-fluorocytosine (5-FC) conversion to active drug 5-fluorouracil by yCD, and has demonstrated therapeutic efficacy in preclinical and clinical trials of various cancers.. Toca 511/5-FC treatment may also induce antitumor immunity.. Here, we first examined antitumor immune responses activated by Toca 511/5-FC treatment in an immunocompetent murine pancreatic cancer model. We then evaluated the therapeutic effects achieved in combination with anti-programmed cell death protein 1 antibody.

    Article Title: Monocyte infiltration induces CNS arginine catabolism to fuel neuroinflammation.
    Article Snippet: .. Flow cytometry and cell sorting For analysis of spinal cord cell populations, stainings were performed in PBS using fixable viability dye (Invitrogen, 65-0865-14, 1:2,000, 10 min at room temperature) as well as the following antibodies: CD45.2-BV650 (clone 104, Biolegend, 109836, 1:100), Ly6G-BV605 (clone 1A8, Biolegend, 127639, 1:100), Ly6G-AF700 (clone 1A8, Biolegend, 127622, 1:100), CD3-BV605 (clone 145-2C11, Biolegend, 100351, 1:100), Ly6C-BV605 (clone HK14, Biolegend, 128035, 1:100), Ly6C-BV510 (clone HK14, Biolegend, 128033, 1:100), CD11b-BV510 (clone M1/70, Biolegend, 101245, 1:100), CD11b-PB (clone M1/70.15, Invitrogen, RM2828, 1:100), CD11b-BV605 (clone M1/70, Biolegend, 101237, 1:100), CX3CR1-PeCy7 (clone SA011F11, Biolegend, 149015, 1:100), CCR2-BV421 (clone SA203G11, Biolegend, 150605, 1:100), CCR2-FITC (clone SA203G11, Biolegend, 150607, 1:100), F4/80-BV510 (clone BM8, Biolegend, 123135, 1:100), F4/80-BV605 (clone BM8, Biolegend, 123133, 1:100), F4/80-FITC (clone BM8, Biolegend, 123108, 1:100), CD11c-PE-Cy5.5 (clone N418, Invitrogen, 35-0114-82, 1:100), P2RY12-APC (clone S16007D, Biolegend, 848006, 1:100), P2RY12-AF488 (clone S16007D, Biolegend, 848016, 1:100), MerTK-APC (clone 2B10C42, Biolegend, 151507, 1:100), CD44-FITC (clone IM7, Invitrogen, 11-0441-82, 1:100), CD44-eF506 (clone IM7, Invitrogen, 69-0441-82, 1:100), CD64-BV605 (clone X545/7.1, Biolegend, 139323, 1:100), CD3-AF700 (clone 17A2, Biolegend, 100216, 1:100), CD3-rF710 (clone 17A2, Tonbo, 80-0032-U100, 1:80), CD8-BV510 (clone 53-6.7, Biolegend, 100751, 1:80), CD4-APC (clone REA604, Miltenyi Biotec, 130-116-487, 1:80), CD4-PE (clone REA604, Miltenyi Biotec, 130-116-509, 1:80) and CD25-BV421 (clone PC61, Biolegend, 102033, 1:80). ..

    Article Title: Patterns of Differentially Expressed circRNAs in Human Thymocytes
    Article Snippet: .. All the immunophenotypes of the thymocyte populations were magnetically sorted and validated by flow cytometry using a FACSCalibur cytometer (Becton Dickinson, San Diego, CA, USA) with the following antibodies: CD34-PE, CD8-APC and CD4-PE (all from MACS Miltenyi Biotec), revealing more than 98% purification efficiency. ..

    Control:

    Article Title: Therapeutic activity of retroviral replicating vector-mediated gene therapy in combination with anti-PD-1 antibody in a murine pancreatic cancer model.
    Article Snippet: Toca 511, a tumor-selective retroviral replicating vector encoding the yeast cytosine deaminase (yCD) gene, exerts direct antitumor effects through intratumoral prodrug 5-fluorocytosine (5-FC) conversion to active drug 5-fluorouracil by yCD, and has demonstrated therapeutic efficacy in preclinical and clinical trials of various cancers.. Toca 511/5-FC treatment may also induce antitumor immunity.. Here, we first examined antitumor immune responses activated by Toca 511/5-FC treatment in an immunocompetent murine pancreatic cancer model. We then evaluated the therapeutic effects achieved in combination with anti-programmed cell death protein 1 antibody.

    FACS:

    Article Title: Monocyte infiltration induces CNS arginine catabolism to fuel neuroinflammation.
    Article Snippet: .. Flow cytometry and cell sorting For analysis of spinal cord cell populations, stainings were performed in PBS using fixable viability dye (Invitrogen, 65-0865-14, 1:2,000, 10 min at room temperature) as well as the following antibodies: CD45.2-BV650 (clone 104, Biolegend, 109836, 1:100), Ly6G-BV605 (clone 1A8, Biolegend, 127639, 1:100), Ly6G-AF700 (clone 1A8, Biolegend, 127622, 1:100), CD3-BV605 (clone 145-2C11, Biolegend, 100351, 1:100), Ly6C-BV605 (clone HK14, Biolegend, 128035, 1:100), Ly6C-BV510 (clone HK14, Biolegend, 128033, 1:100), CD11b-BV510 (clone M1/70, Biolegend, 101245, 1:100), CD11b-PB (clone M1/70.15, Invitrogen, RM2828, 1:100), CD11b-BV605 (clone M1/70, Biolegend, 101237, 1:100), CX3CR1-PeCy7 (clone SA011F11, Biolegend, 149015, 1:100), CCR2-BV421 (clone SA203G11, Biolegend, 150605, 1:100), CCR2-FITC (clone SA203G11, Biolegend, 150607, 1:100), F4/80-BV510 (clone BM8, Biolegend, 123135, 1:100), F4/80-BV605 (clone BM8, Biolegend, 123133, 1:100), F4/80-FITC (clone BM8, Biolegend, 123108, 1:100), CD11c-PE-Cy5.5 (clone N418, Invitrogen, 35-0114-82, 1:100), P2RY12-APC (clone S16007D, Biolegend, 848006, 1:100), P2RY12-AF488 (clone S16007D, Biolegend, 848016, 1:100), MerTK-APC (clone 2B10C42, Biolegend, 151507, 1:100), CD44-FITC (clone IM7, Invitrogen, 11-0441-82, 1:100), CD44-eF506 (clone IM7, Invitrogen, 69-0441-82, 1:100), CD64-BV605 (clone X545/7.1, Biolegend, 139323, 1:100), CD3-AF700 (clone 17A2, Biolegend, 100216, 1:100), CD3-rF710 (clone 17A2, Tonbo, 80-0032-U100, 1:80), CD8-BV510 (clone 53-6.7, Biolegend, 100751, 1:80), CD4-APC (clone REA604, Miltenyi Biotec, 130-116-487, 1:80), CD4-PE (clone REA604, Miltenyi Biotec, 130-116-509, 1:80) and CD25-BV421 (clone PC61, Biolegend, 102033, 1:80). ..

    Cytometry:

    Article Title: Patterns of Differentially Expressed circRNAs in Human Thymocytes
    Article Snippet: .. All the immunophenotypes of the thymocyte populations were magnetically sorted and validated by flow cytometry using a FACSCalibur cytometer (Becton Dickinson, San Diego, CA, USA) with the following antibodies: CD34-PE, CD8-APC and CD4-PE (all from MACS Miltenyi Biotec), revealing more than 98% purification efficiency. ..

    Magnetic Cell Separation:

    Article Title: Patterns of Differentially Expressed circRNAs in Human Thymocytes
    Article Snippet: .. All the immunophenotypes of the thymocyte populations were magnetically sorted and validated by flow cytometry using a FACSCalibur cytometer (Becton Dickinson, San Diego, CA, USA) with the following antibodies: CD34-PE, CD8-APC and CD4-PE (all from MACS Miltenyi Biotec), revealing more than 98% purification efficiency. ..

    Purification:

    Article Title: Patterns of Differentially Expressed circRNAs in Human Thymocytes
    Article Snippet: .. All the immunophenotypes of the thymocyte populations were magnetically sorted and validated by flow cytometry using a FACSCalibur cytometer (Becton Dickinson, San Diego, CA, USA) with the following antibodies: CD34-PE, CD8-APC and CD4-PE (all from MACS Miltenyi Biotec), revealing more than 98% purification efficiency. ..



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    Miltenyi Biotec cd4 pe vio 770
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    Cd4 Pe Vio 770, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Miltenyi Biotec cd4 pe
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    94
    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 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Flow immunophenotyping release testing data are presented in A-G and IFNγ ELISpot data presented in H-K (Arm A: n = 12; Arms B/C: n = 27). Each data point represents a separate TAA-T cell product infused into patients (biological replicates). Lower whisker is defined as the minima, upper whisker is defined as the maxima, center is defined as median, lower bound of box is defined as Q1 (25th percentile), and upper bound of box is defined as Q3 (75th percentile). a ) % Total T cells (CD3 + ). Arm A: Minima=51, Maxima=99, Median=97, 25th Percentile=95, 75th Percentile=99; Arms B/C: Minima=31, Maxima=100, Median=97, 25th Percentile=94, 75th Percentile=98. b ) % CD4 + T cells (CD3 + CD4+ of CD45 + ). Arm A: Minima=0.50, Maxima=88, Median=16, 25th Percentile=2.6, 75th Percentile=38; Arms B/C: Minima=0.40, Maxima=55, Median=9.9, 25th Percentile=6.1, 75th Percentile=17. c ) %CD8 + T cells (% CD3 + CD8+ of CD45 + ). Arm A: Minima=4.3, Maxima=88, Median=37, 25th Percentile=22, 75th Percentile=47; Arms B/C: Minima=19, Maxima=91, Median=51, 25th Percentile=36, 75th Percentile=71. d ) % αβ T cells (% TCRαβ+ of CD3 + ). Arm A: Minima=8.2, Maxima=94, Median=69, 25th Percentile=28, 75th Percentile=86; Arms B/C: Minima=32, Maxima=98, Median=71, 25th Percentile=48, 75th Percentile=84. e ) % γδ T cells (% TCRγδ+ of CD3 + ). Arm A: Minima=5.1, Maxima=79, Median=28, 25th Percentile=8.4, 75th Percentile=67; Arms B/C: Minima=1.1, Maxima=83, Median=25, 25th Percentile=10, 75th Percentile=53. f ) % CD3 + CD16 + CD56+ of CD45 + . Arm A: Minima=2.8, Maxima=25, Median=11, 25th Percentile=7.2, 75th Percentile=17; Arms B/C: Minima=3.4, Maxima=64, Median=16, 25th Percentile=8.1, 75th Percentile=28. g ) % NK cells (CD16/CD56 + CD3- of CD45 + ). Arm A: Minima=0.40, Maxima=48, Median=1.0, 25th Percentile=0.48, 75th Percentile=4.6; Arms B/C: Minima=0.10, Maxima=62, Median=1.6, 25th Percentile=0.60, 75th Percentile=6.1. h - i ) IFNγ ELISpot with background correction for Arm A ( h ) and Arms B/C ( i ). Negative values after background normalization have been normalized to zero for visual representation.

    Journal: Nature Medicine

    Article Title: Multi-antigen-targeting T cells in pediatric central nervous system tumors: a phase 1 trial

    doi: 10.1038/s41591-026-04449-9

    Figure Lengend Snippet: Flow immunophenotyping release testing data are presented in A-G and IFNγ ELISpot data presented in H-K (Arm A: n = 12; Arms B/C: n = 27). Each data point represents a separate TAA-T cell product infused into patients (biological replicates). Lower whisker is defined as the minima, upper whisker is defined as the maxima, center is defined as median, lower bound of box is defined as Q1 (25th percentile), and upper bound of box is defined as Q3 (75th percentile). a ) % Total T cells (CD3 + ). Arm A: Minima=51, Maxima=99, Median=97, 25th Percentile=95, 75th Percentile=99; Arms B/C: Minima=31, Maxima=100, Median=97, 25th Percentile=94, 75th Percentile=98. b ) % CD4 + T cells (CD3 + CD4+ of CD45 + ). Arm A: Minima=0.50, Maxima=88, Median=16, 25th Percentile=2.6, 75th Percentile=38; Arms B/C: Minima=0.40, Maxima=55, Median=9.9, 25th Percentile=6.1, 75th Percentile=17. c ) %CD8 + T cells (% CD3 + CD8+ of CD45 + ). Arm A: Minima=4.3, Maxima=88, Median=37, 25th Percentile=22, 75th Percentile=47; Arms B/C: Minima=19, Maxima=91, Median=51, 25th Percentile=36, 75th Percentile=71. d ) % αβ T cells (% TCRαβ+ of CD3 + ). Arm A: Minima=8.2, Maxima=94, Median=69, 25th Percentile=28, 75th Percentile=86; Arms B/C: Minima=32, Maxima=98, Median=71, 25th Percentile=48, 75th Percentile=84. e ) % γδ T cells (% TCRγδ+ of CD3 + ). Arm A: Minima=5.1, Maxima=79, Median=28, 25th Percentile=8.4, 75th Percentile=67; Arms B/C: Minima=1.1, Maxima=83, Median=25, 25th Percentile=10, 75th Percentile=53. f ) % CD3 + CD16 + CD56+ of CD45 + . Arm A: Minima=2.8, Maxima=25, Median=11, 25th Percentile=7.2, 75th Percentile=17; Arms B/C: Minima=3.4, Maxima=64, Median=16, 25th Percentile=8.1, 75th Percentile=28. g ) % NK cells (CD16/CD56 + CD3- of CD45 + ). Arm A: Minima=0.40, Maxima=48, Median=1.0, 25th Percentile=0.48, 75th Percentile=4.6; Arms B/C: Minima=0.10, Maxima=62, Median=1.6, 25th Percentile=0.60, 75th Percentile=6.1. h - i ) IFNγ ELISpot with background correction for Arm A ( h ) and Arms B/C ( i ). Negative values after background normalization have been normalized to zero for visual representation.

    Article Snippet: An FcR blocking agent (Miltenyi Biotec, cat. no. 130-059-901) was added to the tubes for 5 min to prevent any nonspecific binding, followed by an appropriate antibody mix (shown in tables below), which was incubated in the dark for 15 min. Antibodies for the pan leukocyte panel included CD14 Vio Blue (Miltenyi Biotec, cat. no. 130-110-524), CD19 FITC (Miltenyi Biotec, cat. no. 130-113-646), CD16 PE (Miltenyi Biotec, cat. no. 130-113-393), CD56 PE (Miltenyi Biotec, cat. no. 130-113-312), CD3 Per CP Vio-770 (Miltenyi Biotec, cat. no. 130-113-141), CD4 PE-Vio-770 (Miltenyi Biotec, cat. no. 130-113-227), CD45 APC (Miltenyi Biotec, cat. no. 130-110-633) and CD8 APC Vio-770 (Miltenyi Biotec, cat. no. 130-110-681).

    Techniques: Enzyme-linked Immunospot, Whisker Assay

    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