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Cell Signaling Technology Inc cd8a c8 144b mouse mab cell signaling technology

Figure 1. Anti-TGF-b CAR T cells reduced tumor growth and promoted T cell expansion in vivo (A) Anti-TGF-b CAR vector (T28zT2), anti-GPC3 CAR vector (G28zT2), and anti-CD19 CAR vector (1928zT2) based on an anti-TGF-b scFv (US20140127230A1), anti-GPC3 scFv (GC33), or anti-CD19 scFv (FMC63), respectively. All contained expression cassettes encoding a human <t>CD8</t> leader signal peptide, CD28, CD3z, and TLR2 signaling domain along with eGFP using 2A self-cleaving peptide (2A). The eGFP expression was used to monitor CAR-transduced cells. (B) The percentage of Huh7 cells with 1928zT2, G28zT2, or T28zT2 T cell-induced lysis after 72 h; data are the mean percentage of tumor cell-speciﬁc lysis ± SEM values; n = 3 independent experiments; two-way ANOVA with Tukey’s multiple comparisons test; ****p % 0.0001.

Cd8a C8 144b Mouse Mab Cell Signaling Technology, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 9025 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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1) Product Images from "CD4 + anti-TGF-β CAR T cells and CD8 + conventional CAR T cells exhibit synergistic antitumor effects."

Article Title: CD4 + anti-TGF-β CAR T cells and CD8 + conventional CAR T cells exhibit synergistic antitumor effects.

Journal: Cell reports. Medicine

doi: 10.1016/j.xcrm.2025.102020

Figure Legend Snippet: Figure 1. Anti-TGF-b CAR T cells reduced tumor growth and promoted T cell expansion in vivo (A) Anti-TGF-b CAR vector (T28zT2), anti-GPC3 CAR vector (G28zT2), and anti-CD19 CAR vector (1928zT2) based on an anti-TGF-b scFv (US20140127230A1), anti-GPC3 scFv (GC33), or anti-CD19 scFv (FMC63), respectively. All contained expression cassettes encoding a human CD8 leader signal peptide, CD28, CD3z, and TLR2 signaling domain along with eGFP using 2A self-cleaving peptide (2A). The eGFP expression was used to monitor CAR-transduced cells. (B) The percentage of Huh7 cells with 1928zT2, G28zT2, or T28zT2 T cell-induced lysis after 72 h; data are the mean percentage of tumor cell-speciﬁc lysis ± SEM values; n = 3 independent experiments; two-way ANOVA with Tukey’s multiple comparisons test; ****p % 0.0001.

Techniques Used: In Vivo, Plasmid Preparation, Expressing, Lysis

Figure 3. CD4+ but not CD8+ T28zT2 T cells are effective for tumor growth inhibition (A) CD4+ and CD8+ T cell compartments were sorted, and CD8+ T28zT2 T cells and CD4+ T28zT2 cells were generated. We also mixed the CD4+ and CD8+ T28zT2 T cells at a ratio of 1:1 to obtain CD3+ T28zT2 T cells. We then infused a total of 5 3 106 of the three types of T28zT2 T cells, CD3+ 1928zT2 T cells, or CD3+ M28zT2 T cells peritumorally into subcutaneous xenografts of NSI mice inoculated with 1 3 106 AsPc-1 cells (day 0). Tumor volumes were monitored on the indicated days

Figure Legend Snippet: Figure 3. CD4+ but not CD8+ T28zT2 T cells are effective for tumor growth inhibition (A) CD4+ and CD8+ T cell compartments were sorted, and CD8+ T28zT2 T cells and CD4+ T28zT2 cells were generated. We also mixed the CD4+ and CD8+ T28zT2 T cells at a ratio of 1:1 to obtain CD3+ T28zT2 T cells. We then infused a total of 5 3 106 of the three types of T28zT2 T cells, CD3+ 1928zT2 T cells, or CD3+ M28zT2 T cells peritumorally into subcutaneous xenografts of NSI mice inoculated with 1 3 106 AsPc-1 cells (day 0). Tumor volumes were monitored on the indicated days

Techniques Used: Inhibition, Generated

Figure 5. A combination of CD4+ anti-TGF-b CAR T cells and CD8+ anti-MSLN CAR T cells exhibits augmented antitumor effects in NSCLC PDX (A) NSCLC PDX tumors were diced into 30 mm3 pieces, and tissue were inoculated subcutaneously into the right ﬂanks of 8-week-old male NSI mice. 5 3 106

Figure Legend Snippet: Figure 5. A combination of CD4+ anti-TGF-b CAR T cells and CD8+ anti-MSLN CAR T cells exhibits augmented antitumor effects in NSCLC PDX (A) NSCLC PDX tumors were diced into 30 mm3 pieces, and tissue were inoculated subcutaneously into the right ﬂanks of 8-week-old male NSI mice. 5 3 106

Techniques Used:

Figure 6. TGF-b1 suppressed OXPHOS activity in CD4+ human T cells (A–D) CD4+ and CD8+ T cells (1 3 106) were activated with CD3/CD28 mAbs for 24 h, followed by PBS or TGF-b1 (10 ng/mL) treatment for 16 h. (A) Mitochondrial morphology of CD4+ and CD8+ T cells upon PBS or TGF-b1 (10 ng/mL) treatment, as determined by spinning disk confocal microscopy. Mitochondria are green (MitoTracker Green), and nuclei are blue (DAPI). Scale bar, 5 mm. (B) Relative lengths of mitochondria, as analyzed by ImageJ software, in CD4+ and CD8+ T cells (2 independent experiments). Each dot represents the mean relative length of the mitochondria in a sample. Data are shown as the mean ± SEM values; paired two-tailed t test; ****p % 0.0001. (C) Immunoblot analysis of cellular protein extracts probed with antibodies against pSMAD2S465/467 (top)/pSMAD3S423/425

Figure Legend Snippet: Figure 6. TGF-b1 suppressed OXPHOS activity in CD4+ human T cells (A–D) CD4+ and CD8+ T cells (1 3 106) were activated with CD3/CD28 mAbs for 24 h, followed by PBS or TGF-b1 (10 ng/mL) treatment for 16 h. (A) Mitochondrial morphology of CD4+ and CD8+ T cells upon PBS or TGF-b1 (10 ng/mL) treatment, as determined by spinning disk confocal microscopy. Mitochondria are green (MitoTracker Green), and nuclei are blue (DAPI). Scale bar, 5 mm. (B) Relative lengths of mitochondria, as analyzed by ImageJ software, in CD4+ and CD8+ T cells (2 independent experiments). Each dot represents the mean relative length of the mitochondria in a sample. Data are shown as the mean ± SEM values; paired two-tailed t test; ****p % 0.0001. (C) Immunoblot analysis of cellular protein extracts probed with antibodies against pSMAD2S465/467 (top)/pSMAD3S423/425

Techniques Used: Activity Assay, Confocal Microscopy, Software, Two Tailed Test, Western Blot

Recombinant:

Article Title: CD4 + anti-TGF-β CAR T cells and CD8 + conventional CAR T cells exhibit synergistic antitumor effects.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Human CD25 Antibody R&D systems Cat# AF-223-NA RRID: AB_354408 Purified anti-human CXCR3 (Maxpar Ready) Antibody Fluidigm Cat# 3163004B Purified anti-human CCR6 (Maxpar Ready) Antibody Fluidigm Cat# 3141003A Anti-Human CD45 (HI30)-89Y Fluidigm Cat# 3089003B TIGIT Monoclonal Antibody (MBSA43), Functional Grade, eBioscienceTM Thermo Cat# 16-9500-82 RRID: AB_10718831 FOXP3 Monoclonal Antibody (PCH101), Functional Grade, eBioscienceTM Thermo Cat# 14-4776-82 RRID: AB_467553 APC/Cyanine7 anti-human CD279 (PD-1) Antibody Biolegend Cat# 367416 RRID: AB_2616744 APC anti-human CD279 (PD-1) Antibody Biolegend Cat# 367406 RRID: AB_2566067 PE/Cyanine7 anti-human CD223 (LAG-3) Antibody Biolegend Cat# 369208 RRID: AB_2629835 PerCP/Cyanine5.5 anti-human CD223 (LAG-3) Antibody Biolegend Cat# 369216 RRID: AB_2910413 APC anti-human CD3 Antibody Biolegend Cat# 300439 RRID: AB_2562045 PE/Cyanine7 anti-human CD3 Antibody Biolegend Cat# 300420 RRID: AB_439781 APC/Cyanine7 anti-human CD4 Antibody Biolegend Cat# 317418 RRID: AB_571947 PE anti-human CD8 Antibody Biolegend Cat# 303804 RRID: AB_2860786 APC anti-human CD25 Antibody Biolegend Cat# 356110 RRID: AB_2561977 PE/Cyanine7 anti-human CD69 Antibody Biolegend Cat# 310912 RRID: AB_314847 APC anti-human CD69 Antibody Biolegend Cat# 310910 RRID: AB_314845 PE anti-human CD69 Antibody Biolegend Cat# 310906 RRID: AB_314841 PE anti-human NKG2D Antibody Biolegend Cat# 320806 RRID: AB_492960 APC anti-human MICA/B Antibody Biolegend Cat# 320908 RRID: AB_493196 Human ULBP-2/5/6 APC-conjugated Antibody R&D systems Cat# FAB1298A RRID: AB_2257142 Human Mesothelin APC-conjugated Antibody R&D systems Cat# FAB32652A RRID: AB_2298058 APC Mouse IgG1, k Isotype Ctrl Antibody Biolegend Cat# 400120 RRID: AB_2888687 APC anti-mouse IgG2a Antibody Biolegend Cat# 407110 RRID: AB_2561754 SMAD2/3(D7G7) XP Rabbit mAb Cell Signaling Technology Cat# 8685 RRID: 10891619 Phospho-SMAD2(Ser465/467)/SMAD3 (Ser423/425) (D27F4) Rabbit mAb Cell Signaling Technology Cat# 8828 RRID: 2631089 DRP1 (D6C7) Rabbit mAb Cell Signaling Technology Cat# 8570 RRID: AB_10950498 (Continued on next page) e2 Cell Reports Medicine 6, 102020, March 18, 2025 .. REAGENT or RESOURCE SOURCE IDENTIFIER Phospho-DRP1 (Ser616) (D9A1) Rabbit mAb Cell Signaling Technology Cat# 4494 RRID: AB_11178659 OPA1 (D7C1A) Rabbit mAb Cell Signaling Technology Cat# 67589 RRID: AB_2799728 MFF (E5W4M) XP Rabbit mAb Cell Signaling Technology Cat# 84580 RRID: AB_2799819 SMAD4 (D3R4N) XP Rabbit mAb Cell Signaling Technology Cat# 46535 RRID: AB_2736998 SMAD4 Polyclonal antibody Proteintech Cat# 10231-1-AP RRID: AB_2193323 a-Smooth Muscle Actin (D4K9N) XP Rabbit mAb Cell Signaling Technology Cat# 19245 RRID: AB_2734735 Cleaved Caspase-3 (Asp175) (5A1E) Rabbit mAb Cell Signaling Technology Cat# 9664 RRID: 10828837 CD8a (C8/144B) Mouse mAb Cell Signaling Technology Cat# 70306 RRID: AB_2799781 Anti-CD4 antibody Abcam Cat# ab133616 RRID: AB_2750883 Goat Anti-Rabbit IgG H&L (Alexa Fluor 568) Abcam Cat# ab175471 RRID: AB_2576207 Goat Anti-Mouse IgG H&L (Alexa Fluor 647) Abcam Cat# ab150115 RRID: AB_2687948 Biological samples Patient-derived xenografts (PDX) Jiang. et al.,26 Qin. et al.46 N/A Heathy PBMCs Guangzhou Tianhe Noah Biological Engineering Co., LTD NA Chemicals, peptides, and recombinant proteins TGF beta 1 Protein, Human, Rhesus, Cynomolgus, Canine, Recombinant SinoBiological CAS number: 10804-HNAC TMRM MedChemExpress Cat# HY-D0984 MitoTracker Green Yeasen Cat# 40742ES50 MitoTracker Deep Red Yeasen Cat# 40743ES50 GenCRISPRTM Cas9 v1.2 GenScript Cat# Z03702-1 Seahorse XF Cell Mito Stress Test Kit Agilent Cat# 103010-100 Critical commercial assays Human Granzyme B Precoated ELISA Kit DAKEWE Cat# 1118503 Human IFN-g Precoated ELISA Kit DAKEWE Cat# 1110003 Human IL-2 Precoated ELISA Kit DAKEWE Cat# 1110203 Human TGFb1 Precoated ELISA Kit DAKEWE Cat# 1117102 Human PRF1 Precoated ELISA Kit Bioswamp Cat# HM10300 MACS Pan T cell Isolation Kit Miltenyi Biotec Cat# 130-096-535 MACS GMP T cell TransAct Miltenyi Biotec Cat# 170-076-156 Deposited data Original data of the bulk RNAseq This paper GSA-human: HRA001397 Original western blot images This paper https://doi.org/10.17632/n7htzcxmw4.1 Links: https://data.mendeley.com/drafts/ n7htzcxmw4/1 Experimental models: Cell lines Huh7 Procell Cat# CL-0120 HepG2 Procell Cat# CL-0103 (Continued on next page) Cell Reports Medicine 6, 102020, March 18, 2025 e3 .. REAGENT or RESOURCE SOURCE IDENTIFIER SK-Hep-1 Procell Cat# CL-0212 AsPc-1 Procell Cat# CL-0027 Hela Procell Cat# CL-0292 Jukrat Procell Cat# CL-0315 Experimental models: Organisms/strains Mouse: NSI, NOD-SCID; IL2rg / Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, China N/A Oligonucleotides GZMB target gRNA: 50-TGTCTGCCCTGGCTTCACCT-30 GenScript Cat# C080C988G0 IFNG target gRNA: 50-AAAGAGTGTGGAGACCATCA -30 GenScript Cat# C5095GJ130 ctrl target gRNA: 50-CCGGGTCTTCGAGAAGACCT-3-30 GenScript Cat# C2639HE050 Recombinant DNA Anti-TGFb scFv-CD28 CD3z-2A-eGFP This paper N/A Anti-TGFb scFv-CD28 CD3z-TLR2-2AeGFP This paper N/A Anti-CD19 scFv-CD28 CD3z-TLR2-2AeGFP This paper N/A Anti-GPC3 scFv-CD28 CD3z-TLR2-2AeGFP This paper N/A Anti-MSLN scFv-CD28 CD3z-TLR2-2AeGFP This paper N/A DnTGFbRII-2A-eGFP This paper N/A Anti-TGFb scFv-mCD28-mCD3z-mTLR22A-eGFP This paper N/A Anti-CD19 scFv-mCD28-mCD3z-mTLR22A-eGFP This paper N/A Software and algorithms FlowJo v10 TreeStar https://www.flowjo.com/solutions/flowjo/ downloads Prism 9.0 GraphPad https://www.graphpad.com/ scientificsoftware/prism/ ImageJ National Institutes of Health https://imagej.en.softonic.com BioRender BioRender https://www.biorender.com/ BGI Dr. Tom BGI https://biosys.bgi.com/ iDEP iDEP http://bioinformatics.sdstate.edu/idep/

Enzyme-linked Immunosorbent Assay:

Magnetic Cell Separation:

Cell Isolation:

RNA sequencing:

Western Blot:

other:

Article Title: Disease-specific suppressive granulocytes participate in glioma progression.
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