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cd8a microbeads  (Miltenyi Biotec)


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    Miltenyi Biotec cd8a microbeads
    Neonatal and adult RTEs are phenotypically distinct (A) Schematic of the timestamping system marking T cells made at the time of tamoxifen administration. RTEs were collected at 2 weeks post-marking. (B) Left: Representative contour plots displaying virtual memory (VM, CD44 hi CD122 hi ) and true naive (TN, CD44 lo CD122 lo ) <t>CD8</t> + populations. Right: Statistical analysis of VM population within the marked T cell population. (C) Statistical analysis and representative histograms of the RTE markers Qa2 (left) and CD103 (right). (D) Statistical analysis and representative histograms of the phenotypic markers CD127 (left) and CD11a (right). n = 8-9 mice per group from 2 independent experiments. For statistical analysis, unpaired t tests were performed. ns, not significant; ∗∗∗, p < 0.001; ∗∗∗∗, p < 0.0001.
    Cd8a Microbeads, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 96/100, based on 268 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/cd8a/CD8a+(Ly-2)+MicroBeads%2C+mouse/pmc13098507-44-0-4
    Average 96 stars, based on 268 article reviews
    cd8a microbeads - by Bioz Stars, 2026-10
    96/100 stars

    Images

    1) Product Images from "Single-cell gene expression and TCR profiling reveal age-related differences in recent thymic emigrants"

    Article Title: Single-cell gene expression and TCR profiling reveal age-related differences in recent thymic emigrants

    Journal: iScience

    doi: 10.1016/j.isci.2026.115582

    Neonatal and adult RTEs are phenotypically distinct (A) Schematic of the timestamping system marking T cells made at the time of tamoxifen administration. RTEs were collected at 2 weeks post-marking. (B) Left: Representative contour plots displaying virtual memory (VM, CD44 hi CD122 hi ) and true naive (TN, CD44 lo CD122 lo ) CD8 + populations. Right: Statistical analysis of VM population within the marked T cell population. (C) Statistical analysis and representative histograms of the RTE markers Qa2 (left) and CD103 (right). (D) Statistical analysis and representative histograms of the phenotypic markers CD127 (left) and CD11a (right). n = 8-9 mice per group from 2 independent experiments. For statistical analysis, unpaired t tests were performed. ns, not significant; ∗∗∗, p < 0.001; ∗∗∗∗, p < 0.0001.
    Figure Legend Snippet: Neonatal and adult RTEs are phenotypically distinct (A) Schematic of the timestamping system marking T cells made at the time of tamoxifen administration. RTEs were collected at 2 weeks post-marking. (B) Left: Representative contour plots displaying virtual memory (VM, CD44 hi CD122 hi ) and true naive (TN, CD44 lo CD122 lo ) CD8 + populations. Right: Statistical analysis of VM population within the marked T cell population. (C) Statistical analysis and representative histograms of the RTE markers Qa2 (left) and CD103 (right). (D) Statistical analysis and representative histograms of the phenotypic markers CD127 (left) and CD11a (right). n = 8-9 mice per group from 2 independent experiments. For statistical analysis, unpaired t tests were performed. ns, not significant; ∗∗∗, p < 0.001; ∗∗∗∗, p < 0.0001.

    Techniques Used:

    Neonatal RTEs exhibit enhanced immune functionality (A) Schematic of in vitro stimulation. CD8 + T cells from neonatal and adult timestamped (ts) mice were coated with CTV and stimulated for 48 h via antibody-mediated crosslinking of CD3/CD28. (B) Representative histograms of CTV dilution. (C) Division index. Unpaired t test was performed for statistical analysis. n = 8 samples per group from 2 independent experiments. (D) Schematic of the in vivo infection experiment. Thymic lobes from newborn GFP timestamped mice were grafted under the kidney capsule of adult RFP timestamped mice. Tamoxifen was administered to thymic graft recipients, and after 2 weeks, recipients were infected with 5 × 10 3 CFUs of LM-gB. Spleens were collected at 5 dpi. (E) Percentage of timestamped tetramer-positive cells with a short-lived effector phenotype (KLRG1 hi CD127 lo ). (F) Percentage of timestamped tetramer-positive cells with a memory precursor effector phenotype (KLRG1 lo CD127 hi ). (G) Percentage of timestamped cells producing IFN-γ. N = 19. For statistical analysis, paired t tests were performed. ∗∗, p < 0.01; ∗∗∗, p < 0.001; ∗∗∗∗, p < 0.0001.
    Figure Legend Snippet: Neonatal RTEs exhibit enhanced immune functionality (A) Schematic of in vitro stimulation. CD8 + T cells from neonatal and adult timestamped (ts) mice were coated with CTV and stimulated for 48 h via antibody-mediated crosslinking of CD3/CD28. (B) Representative histograms of CTV dilution. (C) Division index. Unpaired t test was performed for statistical analysis. n = 8 samples per group from 2 independent experiments. (D) Schematic of the in vivo infection experiment. Thymic lobes from newborn GFP timestamped mice were grafted under the kidney capsule of adult RFP timestamped mice. Tamoxifen was administered to thymic graft recipients, and after 2 weeks, recipients were infected with 5 × 10 3 CFUs of LM-gB. Spleens were collected at 5 dpi. (E) Percentage of timestamped tetramer-positive cells with a short-lived effector phenotype (KLRG1 hi CD127 lo ). (F) Percentage of timestamped tetramer-positive cells with a memory precursor effector phenotype (KLRG1 lo CD127 hi ). (G) Percentage of timestamped cells producing IFN-γ. N = 19. For statistical analysis, paired t tests were performed. ∗∗, p < 0.01; ∗∗∗, p < 0.001; ∗∗∗∗, p < 0.0001.

    Techniques Used: In Vitro, In Vivo, Infection

    Related Articles

    Cell Isolation:

    Article Title: Stem-like precursors of exhausted Th cells upheld by a Tox-Myb-Eomes transcriptional hierarchy propagate Th cell responses in chronic infection.
    Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Anti-Biotin microbeads Miltenyi Biotec Cat# 130-090-485; RRID: AB_244365 Anti-mouse CD4 Microbead Miltenyi Biotec Cat# 130-117-043 Anti-mouse CD8 Microbead Miltenyi Biotec Cat# 130-117-044 Anti-mouse CD19 Microbead Miltenyi Biotec Cat# 130-121-301 Percoll GE Healthcare Bioscience Cat# 17089101 Peroxidase-conjugated AffiniPure Goat Anti-Rat IgG Jackson ImmunoResearch Laboratories Cat# 112-035-003 Anti-VL-4 antibody BioXcell Cat# BE0106; RRID: AB_10949017 Triton X-100 Ajax Finechem Cat# AJA1552 O-phenylenediamine dihydrochloride Thermo Fisher Scientific Cat# P7288 H2O2 Perhydrol Sigma-Aldrich Cat# 7722-84-1 Na2HPO4 Sigma-Aldrich Cat# 7558-79-4 Citric Acid Sigma-Aldrich Cat# 251275 Critical commercial assays Naı̈ve CD4 T cell isolation kit Miltenyi Biotec Cat# 130-104-453 FOXP3 transcription factor staining buffer set ebioscience Cat# 00-5523-00 Chromium NEXT GEM Single Cell 5’ Kit v2 10x Genomics Cat# PN-1000263 Library Construction Kit 10x Genomics Cat# PN-1000190 Chromium Single Cell Mouse TCR Amplification Kit 10x Genomics Cat# PN-1000254 Dual Index Kit TN Set 10x Genomics Cat# PN-1000215 Dual Index Kit TT Set 10x Genomics Cat# PN-1000250 High Sensitivity D5000 Reagents Agilent Technologies Cat# 5067-5593 Deposited data Single cell-RNA sequencing raw data This paper GEO: GSE283820 Experimental models: Organisms/strains MybGFP Stephen L. Nutt lab, Walter and Eliza Hall Institute of Medical Research N/A Foxp3RFP Stephen L. Nutt lab, Walter and Eliza Hall Institute of Medical Research N/A Mybfl/flCd4Cre Stephen L. Nutt lab, Walter and Eliza Hall Institute of Medical Research N/A Mybfl/flCd4Cre SMARTA Axel Kallies lab, The Peter Doherty Institute, University of Melbourne N/A Mybfl/flERT2Cre Axel Kallies lab, The Peter Doherty Institute, University of Melbourne N/A Eomesfl/flCd4Cre Laura Mackay lab, The Peter Doherty Institute, University of Melbourne N/A ERT2Cre The Jackson laboratory N/A Cd4Cre The Jackson laboratory N/A SMARTA The Jackson laboratory N/A C57BL/6 The Peter Doherty Institute, University of Melbourne N/A Ly5.1 Australian Bio Resources/ Ozgene Animal Resources Centre N/A Oligonucleotides TOX sgRNA Synthego 1.CGAAGATTGTGGCATCAATG 2.AGCGGCCTGCCTCTGATATG Software and algorithms Flowjo 10.10.0 Flowjo, LLC RRID:SCR_008520 (Continued on next page) ll OPEN ACCESSArticle Immunity 59, 1–17.e1–e5, July 14, 2026 e2 ..

    Staining:

    Article Title: Stem-like precursors of exhausted Th cells upheld by a Tox-Myb-Eomes transcriptional hierarchy propagate Th cell responses in chronic infection.
    Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Anti-Biotin microbeads Miltenyi Biotec Cat# 130-090-485; RRID: AB_244365 Anti-mouse CD4 Microbead Miltenyi Biotec Cat# 130-117-043 Anti-mouse CD8 Microbead Miltenyi Biotec Cat# 130-117-044 Anti-mouse CD19 Microbead Miltenyi Biotec Cat# 130-121-301 Percoll GE Healthcare Bioscience Cat# 17089101 Peroxidase-conjugated AffiniPure Goat Anti-Rat IgG Jackson ImmunoResearch Laboratories Cat# 112-035-003 Anti-VL-4 antibody BioXcell Cat# BE0106; RRID: AB_10949017 Triton X-100 Ajax Finechem Cat# AJA1552 O-phenylenediamine dihydrochloride Thermo Fisher Scientific Cat# P7288 H2O2 Perhydrol Sigma-Aldrich Cat# 7722-84-1 Na2HPO4 Sigma-Aldrich Cat# 7558-79-4 Citric Acid Sigma-Aldrich Cat# 251275 Critical commercial assays Naı̈ve CD4 T cell isolation kit Miltenyi Biotec Cat# 130-104-453 FOXP3 transcription factor staining buffer set ebioscience Cat# 00-5523-00 Chromium NEXT GEM Single Cell 5’ Kit v2 10x Genomics Cat# PN-1000263 Library Construction Kit 10x Genomics Cat# PN-1000190 Chromium Single Cell Mouse TCR Amplification Kit 10x Genomics Cat# PN-1000254 Dual Index Kit TN Set 10x Genomics Cat# PN-1000215 Dual Index Kit TT Set 10x Genomics Cat# PN-1000250 High Sensitivity D5000 Reagents Agilent Technologies Cat# 5067-5593 Deposited data Single cell-RNA sequencing raw data This paper GEO: GSE283820 Experimental models: Organisms/strains MybGFP Stephen L. Nutt lab, Walter and Eliza Hall Institute of Medical Research N/A Foxp3RFP Stephen L. Nutt lab, Walter and Eliza Hall Institute of Medical Research N/A Mybfl/flCd4Cre Stephen L. Nutt lab, Walter and Eliza Hall Institute of Medical Research N/A Mybfl/flCd4Cre SMARTA Axel Kallies lab, The Peter Doherty Institute, University of Melbourne N/A Mybfl/flERT2Cre Axel Kallies lab, The Peter Doherty Institute, University of Melbourne N/A Eomesfl/flCd4Cre Laura Mackay lab, The Peter Doherty Institute, University of Melbourne N/A ERT2Cre The Jackson laboratory N/A Cd4Cre The Jackson laboratory N/A SMARTA The Jackson laboratory N/A C57BL/6 The Peter Doherty Institute, University of Melbourne N/A Ly5.1 Australian Bio Resources/ Ozgene Animal Resources Centre N/A Oligonucleotides TOX sgRNA Synthego 1.CGAAGATTGTGGCATCAATG 2.AGCGGCCTGCCTCTGATATG Software and algorithms Flowjo 10.10.0 Flowjo, LLC RRID:SCR_008520 (Continued on next page) ll OPEN ACCESSArticle Immunity 59, 1–17.e1–e5, July 14, 2026 e2 ..

    Single Cell:

    Article Title: Stem-like precursors of exhausted Th cells upheld by a Tox-Myb-Eomes transcriptional hierarchy propagate Th cell responses in chronic infection.
    Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Anti-Biotin microbeads Miltenyi Biotec Cat# 130-090-485; RRID: AB_244365 Anti-mouse CD4 Microbead Miltenyi Biotec Cat# 130-117-043 Anti-mouse CD8 Microbead Miltenyi Biotec Cat# 130-117-044 Anti-mouse CD19 Microbead Miltenyi Biotec Cat# 130-121-301 Percoll GE Healthcare Bioscience Cat# 17089101 Peroxidase-conjugated AffiniPure Goat Anti-Rat IgG Jackson ImmunoResearch Laboratories Cat# 112-035-003 Anti-VL-4 antibody BioXcell Cat# BE0106; RRID: AB_10949017 Triton X-100 Ajax Finechem Cat# AJA1552 O-phenylenediamine dihydrochloride Thermo Fisher Scientific Cat# P7288 H2O2 Perhydrol Sigma-Aldrich Cat# 7722-84-1 Na2HPO4 Sigma-Aldrich Cat# 7558-79-4 Citric Acid Sigma-Aldrich Cat# 251275 Critical commercial assays Naı̈ve CD4 T cell isolation kit Miltenyi Biotec Cat# 130-104-453 FOXP3 transcription factor staining buffer set ebioscience Cat# 00-5523-00 Chromium NEXT GEM Single Cell 5’ Kit v2 10x Genomics Cat# PN-1000263 Library Construction Kit 10x Genomics Cat# PN-1000190 Chromium Single Cell Mouse TCR Amplification Kit 10x Genomics Cat# PN-1000254 Dual Index Kit TN Set 10x Genomics Cat# PN-1000215 Dual Index Kit TT Set 10x Genomics Cat# PN-1000250 High Sensitivity D5000 Reagents Agilent Technologies Cat# 5067-5593 Deposited data Single cell-RNA sequencing raw data This paper GEO: GSE283820 Experimental models: Organisms/strains MybGFP Stephen L. Nutt lab, Walter and Eliza Hall Institute of Medical Research N/A Foxp3RFP Stephen L. Nutt lab, Walter and Eliza Hall Institute of Medical Research N/A Mybfl/flCd4Cre Stephen L. Nutt lab, Walter and Eliza Hall Institute of Medical Research N/A Mybfl/flCd4Cre SMARTA Axel Kallies lab, The Peter Doherty Institute, University of Melbourne N/A Mybfl/flERT2Cre Axel Kallies lab, The Peter Doherty Institute, University of Melbourne N/A Eomesfl/flCd4Cre Laura Mackay lab, The Peter Doherty Institute, University of Melbourne N/A ERT2Cre The Jackson laboratory N/A Cd4Cre The Jackson laboratory N/A SMARTA The Jackson laboratory N/A C57BL/6 The Peter Doherty Institute, University of Melbourne N/A Ly5.1 Australian Bio Resources/ Ozgene Animal Resources Centre N/A Oligonucleotides TOX sgRNA Synthego 1.CGAAGATTGTGGCATCAATG 2.AGCGGCCTGCCTCTGATATG Software and algorithms Flowjo 10.10.0 Flowjo, LLC RRID:SCR_008520 (Continued on next page) ll OPEN ACCESSArticle Immunity 59, 1–17.e1–e5, July 14, 2026 e2 ..

    Amplification:

    Article Title: Stem-like precursors of exhausted Th cells upheld by a Tox-Myb-Eomes transcriptional hierarchy propagate Th cell responses in chronic infection.
    Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Anti-Biotin microbeads Miltenyi Biotec Cat# 130-090-485; RRID: AB_244365 Anti-mouse CD4 Microbead Miltenyi Biotec Cat# 130-117-043 Anti-mouse CD8 Microbead Miltenyi Biotec Cat# 130-117-044 Anti-mouse CD19 Microbead Miltenyi Biotec Cat# 130-121-301 Percoll GE Healthcare Bioscience Cat# 17089101 Peroxidase-conjugated AffiniPure Goat Anti-Rat IgG Jackson ImmunoResearch Laboratories Cat# 112-035-003 Anti-VL-4 antibody BioXcell Cat# BE0106; RRID: AB_10949017 Triton X-100 Ajax Finechem Cat# AJA1552 O-phenylenediamine dihydrochloride Thermo Fisher Scientific Cat# P7288 H2O2 Perhydrol Sigma-Aldrich Cat# 7722-84-1 Na2HPO4 Sigma-Aldrich Cat# 7558-79-4 Citric Acid Sigma-Aldrich Cat# 251275 Critical commercial assays Naı̈ve CD4 T cell isolation kit Miltenyi Biotec Cat# 130-104-453 FOXP3 transcription factor staining buffer set ebioscience Cat# 00-5523-00 Chromium NEXT GEM Single Cell 5’ Kit v2 10x Genomics Cat# PN-1000263 Library Construction Kit 10x Genomics Cat# PN-1000190 Chromium Single Cell Mouse TCR Amplification Kit 10x Genomics Cat# PN-1000254 Dual Index Kit TN Set 10x Genomics Cat# PN-1000215 Dual Index Kit TT Set 10x Genomics Cat# PN-1000250 High Sensitivity D5000 Reagents Agilent Technologies Cat# 5067-5593 Deposited data Single cell-RNA sequencing raw data This paper GEO: GSE283820 Experimental models: Organisms/strains MybGFP Stephen L. Nutt lab, Walter and Eliza Hall Institute of Medical Research N/A Foxp3RFP Stephen L. Nutt lab, Walter and Eliza Hall Institute of Medical Research N/A Mybfl/flCd4Cre Stephen L. Nutt lab, Walter and Eliza Hall Institute of Medical Research N/A Mybfl/flCd4Cre SMARTA Axel Kallies lab, The Peter Doherty Institute, University of Melbourne N/A Mybfl/flERT2Cre Axel Kallies lab, The Peter Doherty Institute, University of Melbourne N/A Eomesfl/flCd4Cre Laura Mackay lab, The Peter Doherty Institute, University of Melbourne N/A ERT2Cre The Jackson laboratory N/A Cd4Cre The Jackson laboratory N/A SMARTA The Jackson laboratory N/A C57BL/6 The Peter Doherty Institute, University of Melbourne N/A Ly5.1 Australian Bio Resources/ Ozgene Animal Resources Centre N/A Oligonucleotides TOX sgRNA Synthego 1.CGAAGATTGTGGCATCAATG 2.AGCGGCCTGCCTCTGATATG Software and algorithms Flowjo 10.10.0 Flowjo, LLC RRID:SCR_008520 (Continued on next page) ll OPEN ACCESSArticle Immunity 59, 1–17.e1–e5, July 14, 2026 e2 ..

    Sequencing:

    Article Title: Stem-like precursors of exhausted Th cells upheld by a Tox-Myb-Eomes transcriptional hierarchy propagate Th cell responses in chronic infection.
    Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Anti-Biotin microbeads Miltenyi Biotec Cat# 130-090-485; RRID: AB_244365 Anti-mouse CD4 Microbead Miltenyi Biotec Cat# 130-117-043 Anti-mouse CD8 Microbead Miltenyi Biotec Cat# 130-117-044 Anti-mouse CD19 Microbead Miltenyi Biotec Cat# 130-121-301 Percoll GE Healthcare Bioscience Cat# 17089101 Peroxidase-conjugated AffiniPure Goat Anti-Rat IgG Jackson ImmunoResearch Laboratories Cat# 112-035-003 Anti-VL-4 antibody BioXcell Cat# BE0106; RRID: AB_10949017 Triton X-100 Ajax Finechem Cat# AJA1552 O-phenylenediamine dihydrochloride Thermo Fisher Scientific Cat# P7288 H2O2 Perhydrol Sigma-Aldrich Cat# 7722-84-1 Na2HPO4 Sigma-Aldrich Cat# 7558-79-4 Citric Acid Sigma-Aldrich Cat# 251275 Critical commercial assays Naı̈ve CD4 T cell isolation kit Miltenyi Biotec Cat# 130-104-453 FOXP3 transcription factor staining buffer set ebioscience Cat# 00-5523-00 Chromium NEXT GEM Single Cell 5’ Kit v2 10x Genomics Cat# PN-1000263 Library Construction Kit 10x Genomics Cat# PN-1000190 Chromium Single Cell Mouse TCR Amplification Kit 10x Genomics Cat# PN-1000254 Dual Index Kit TN Set 10x Genomics Cat# PN-1000215 Dual Index Kit TT Set 10x Genomics Cat# PN-1000250 High Sensitivity D5000 Reagents Agilent Technologies Cat# 5067-5593 Deposited data Single cell-RNA sequencing raw data This paper GEO: GSE283820 Experimental models: Organisms/strains MybGFP Stephen L. Nutt lab, Walter and Eliza Hall Institute of Medical Research N/A Foxp3RFP Stephen L. Nutt lab, Walter and Eliza Hall Institute of Medical Research N/A Mybfl/flCd4Cre Stephen L. Nutt lab, Walter and Eliza Hall Institute of Medical Research N/A Mybfl/flCd4Cre SMARTA Axel Kallies lab, The Peter Doherty Institute, University of Melbourne N/A Mybfl/flERT2Cre Axel Kallies lab, The Peter Doherty Institute, University of Melbourne N/A Eomesfl/flCd4Cre Laura Mackay lab, The Peter Doherty Institute, University of Melbourne N/A ERT2Cre The Jackson laboratory N/A Cd4Cre The Jackson laboratory N/A SMARTA The Jackson laboratory N/A C57BL/6 The Peter Doherty Institute, University of Melbourne N/A Ly5.1 Australian Bio Resources/ Ozgene Animal Resources Centre N/A Oligonucleotides TOX sgRNA Synthego 1.CGAAGATTGTGGCATCAATG 2.AGCGGCCTGCCTCTGATATG Software and algorithms Flowjo 10.10.0 Flowjo, LLC RRID:SCR_008520 (Continued on next page) ll OPEN ACCESSArticle Immunity 59, 1–17.e1–e5, July 14, 2026 e2 ..

    Software:

    Article Title: Stem-like precursors of exhausted Th cells upheld by a Tox-Myb-Eomes transcriptional hierarchy propagate Th cell responses in chronic infection.
    Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Anti-Biotin microbeads Miltenyi Biotec Cat# 130-090-485; RRID: AB_244365 Anti-mouse CD4 Microbead Miltenyi Biotec Cat# 130-117-043 Anti-mouse CD8 Microbead Miltenyi Biotec Cat# 130-117-044 Anti-mouse CD19 Microbead Miltenyi Biotec Cat# 130-121-301 Percoll GE Healthcare Bioscience Cat# 17089101 Peroxidase-conjugated AffiniPure Goat Anti-Rat IgG Jackson ImmunoResearch Laboratories Cat# 112-035-003 Anti-VL-4 antibody BioXcell Cat# BE0106; RRID: AB_10949017 Triton X-100 Ajax Finechem Cat# AJA1552 O-phenylenediamine dihydrochloride Thermo Fisher Scientific Cat# P7288 H2O2 Perhydrol Sigma-Aldrich Cat# 7722-84-1 Na2HPO4 Sigma-Aldrich Cat# 7558-79-4 Citric Acid Sigma-Aldrich Cat# 251275 Critical commercial assays Naı̈ve CD4 T cell isolation kit Miltenyi Biotec Cat# 130-104-453 FOXP3 transcription factor staining buffer set ebioscience Cat# 00-5523-00 Chromium NEXT GEM Single Cell 5’ Kit v2 10x Genomics Cat# PN-1000263 Library Construction Kit 10x Genomics Cat# PN-1000190 Chromium Single Cell Mouse TCR Amplification Kit 10x Genomics Cat# PN-1000254 Dual Index Kit TN Set 10x Genomics Cat# PN-1000215 Dual Index Kit TT Set 10x Genomics Cat# PN-1000250 High Sensitivity D5000 Reagents Agilent Technologies Cat# 5067-5593 Deposited data Single cell-RNA sequencing raw data This paper GEO: GSE283820 Experimental models: Organisms/strains MybGFP Stephen L. Nutt lab, Walter and Eliza Hall Institute of Medical Research N/A Foxp3RFP Stephen L. Nutt lab, Walter and Eliza Hall Institute of Medical Research N/A Mybfl/flCd4Cre Stephen L. Nutt lab, Walter and Eliza Hall Institute of Medical Research N/A Mybfl/flCd4Cre SMARTA Axel Kallies lab, The Peter Doherty Institute, University of Melbourne N/A Mybfl/flERT2Cre Axel Kallies lab, The Peter Doherty Institute, University of Melbourne N/A Eomesfl/flCd4Cre Laura Mackay lab, The Peter Doherty Institute, University of Melbourne N/A ERT2Cre The Jackson laboratory N/A Cd4Cre The Jackson laboratory N/A SMARTA The Jackson laboratory N/A C57BL/6 The Peter Doherty Institute, University of Melbourne N/A Ly5.1 Australian Bio Resources/ Ozgene Animal Resources Centre N/A Oligonucleotides TOX sgRNA Synthego 1.CGAAGATTGTGGCATCAATG 2.AGCGGCCTGCCTCTGATATG Software and algorithms Flowjo 10.10.0 Flowjo, LLC RRID:SCR_008520 (Continued on next page) ll OPEN ACCESSArticle Immunity 59, 1–17.e1–e5, July 14, 2026 e2 ..

    Isolation:

    Article Title: Synergistic immunotherapy of NSCLC enabled by a biomimetic nanodelivery system for autophagy-mediated MDSC reprogramming and metabolic modulation.
    Article Snippet: The immunosuppressive tumor microenvironment (TME) in non-small cell lung cancer (NSCLC), particularly the dominance of myeloid-derived suppressor cells (MDSCs), remains a major barrier to effective immunotherapy.. Here, we developed a bioengineered nanoplatform (PD-L1-SSD@NPs) by conjugating an anti-PD-L1 antibody to hollow mesoporous silica nanoparticles for the targeted delivery of saikosaponin D (SSD) to PD-L1-high MDSCs.. This strategy nanoplatform enhanced SSD delivery and activated autophagic flux in MDSCs, thereby reprogramming them from an immunosuppressive phenotype toward an immune-activating state.

    Activation Assay:

    Article Title: Synergistic immunotherapy of NSCLC enabled by a biomimetic nanodelivery system for autophagy-mediated MDSC reprogramming and metabolic modulation.
    Article Snippet: The immunosuppressive tumor microenvironment (TME) in non-small cell lung cancer (NSCLC), particularly the dominance of myeloid-derived suppressor cells (MDSCs), remains a major barrier to effective immunotherapy.. Here, we developed a bioengineered nanoplatform (PD-L1-SSD@NPs) by conjugating an anti-PD-L1 antibody to hollow mesoporous silica nanoparticles for the targeted delivery of saikosaponin D (SSD) to PD-L1-high MDSCs.. This strategy nanoplatform enhanced SSD delivery and activated autophagic flux in MDSCs, thereby reprogramming them from an immunosuppressive phenotype toward an immune-activating state.

    Selection:

    Article Title: Synergistic immunotherapy of NSCLC enabled by a biomimetic nanodelivery system for autophagy-mediated MDSC reprogramming and metabolic modulation.
    Article Snippet: The immunosuppressive tumor microenvironment (TME) in non-small cell lung cancer (NSCLC), particularly the dominance of myeloid-derived suppressor cells (MDSCs), remains a major barrier to effective immunotherapy.. Here, we developed a bioengineered nanoplatform (PD-L1-SSD@NPs) by conjugating an anti-PD-L1 antibody to hollow mesoporous silica nanoparticles for the targeted delivery of saikosaponin D (SSD) to PD-L1-high MDSCs.. This strategy nanoplatform enhanced SSD delivery and activated autophagic flux in MDSCs, thereby reprogramming them from an immunosuppressive phenotype toward an immune-activating state.

    Magnetic Cell Separation:

    Article Title: Synergistic immunotherapy of NSCLC enabled by a biomimetic nanodelivery system for autophagy-mediated MDSC reprogramming and metabolic modulation.
    Article Snippet: The immunosuppressive tumor microenvironment (TME) in non-small cell lung cancer (NSCLC), particularly the dominance of myeloid-derived suppressor cells (MDSCs), remains a major barrier to effective immunotherapy.. Here, we developed a bioengineered nanoplatform (PD-L1-SSD@NPs) by conjugating an anti-PD-L1 antibody to hollow mesoporous silica nanoparticles for the targeted delivery of saikosaponin D (SSD) to PD-L1-high MDSCs.. This strategy nanoplatform enhanced SSD delivery and activated autophagic flux in MDSCs, thereby reprogramming them from an immunosuppressive phenotype toward an immune-activating state.



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    Proangiogenic and immunosuppression phenotype of TANs in the CRLM microenvironment. A, Heatmap showing signature scores for angiogenesis, myeloid cell recruitment, neutrophil cytotoxicity, MDSC, and NET across each TAN and PMN subset. B, Comparison of angiogenesis, myeloid cell recruitment, MDSC, and NET signature scores among TAN and PMN subsets. C, Dot plot illustrating expression levels of major cytokines and chemokines in each TAN and PMN subset. D, Representative IHC images of CD31 staining in CRLM tissues. Scale bars, 50 µm. E, Correlation analysis between CD31 and TAN1 density using Spearman’s rank correlation ( n = 102). F, Representative images and quantification of tube formation in HUVECs when cocultured with conditioned medium from human LGALS3 − neutrophils or LGALS3 + neutrophils ( n = 6). G, UMAP plot showing six monocyte/macrophage subsets across colorectal cancer, adjacent liver tissues, CRLM, and PB samples. H, Shell-like plot depicting ligand–receptor interaction strength between TAN1_LGASL3 and neutrophil/monocyte/macrophage subset in CRLM. I, Selected ligand–receptor interactions between TAN1_LGASL3 and neutrophil/monocyte/macrophage subset in CRLM. J, Representative mIHC images of CD68, CCL3L1, CD66b, CCR1, and BHLHE40 in CRLM. Scale bars, 50 µm. K, Representative images of macrophage migration assays and effects of conditioned medium from human LGALS3 + neutrophils on migration of macrophages (induced from THP-1 cells) in the presence of CCR1 inhibitor (BX471) or anti-CCL3 antibody ( n = 6). L, Comparison of immunosuppressive macrophage signature scores between colorectal cancer and CRLM. M, Schematic of the CRLM mouse model. C57BL/6 mice received intrasplenic injections of mouse colorectal cancer cells (MC38). Mice were subsequently treated with IgG control (IgG; n = 6), anti-Ly6G antibody (anti-Ly6G; n = 6), or anti-Ly6G plus <t>anti-CD8a</t> antibodies (anti-Ly6G + anti-CD8a; n = 6). Livers were harvested on day 14. N, Representative images of liver metastases from IgG, anti-Ly6G, and anti-Ly6G + anti-CD8a groups. O, Comparison of liver weights among the three groups. P values were calculated by the Wilcoxon rank-sum test ( B and L ) and unpaired Student t test ( F , K , and O ). *, P < 0.05; **, P < 0.01; ***, P < 0.001. CRC, colorectal cancer; Ctrl, control.
    Gb21303 Apc Anti Mouse Cd8a Antibody Proteintech, supplied by Proteintech, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    anti cd8a  (Bio X Cell)
    95
    Bio X Cell anti cd8a
    Proangiogenic and immunosuppression phenotype of TANs in the CRLM microenvironment. A, Heatmap showing signature scores for angiogenesis, myeloid cell recruitment, neutrophil cytotoxicity, MDSC, and NET across each TAN and PMN subset. B, Comparison of angiogenesis, myeloid cell recruitment, MDSC, and NET signature scores among TAN and PMN subsets. C, Dot plot illustrating expression levels of major cytokines and chemokines in each TAN and PMN subset. D, Representative IHC images of CD31 staining in CRLM tissues. Scale bars, 50 µm. E, Correlation analysis between CD31 and TAN1 density using Spearman’s rank correlation ( n = 102). F, Representative images and quantification of tube formation in HUVECs when cocultured with conditioned medium from human LGALS3 − neutrophils or LGALS3 + neutrophils ( n = 6). G, UMAP plot showing six monocyte/macrophage subsets across colorectal cancer, adjacent liver tissues, CRLM, and PB samples. H, Shell-like plot depicting ligand–receptor interaction strength between TAN1_LGASL3 and neutrophil/monocyte/macrophage subset in CRLM. I, Selected ligand–receptor interactions between TAN1_LGASL3 and neutrophil/monocyte/macrophage subset in CRLM. J, Representative mIHC images of CD68, CCL3L1, CD66b, CCR1, and BHLHE40 in CRLM. Scale bars, 50 µm. K, Representative images of macrophage migration assays and effects of conditioned medium from human LGALS3 + neutrophils on migration of macrophages (induced from THP-1 cells) in the presence of CCR1 inhibitor (BX471) or anti-CCL3 antibody ( n = 6). L, Comparison of immunosuppressive macrophage signature scores between colorectal cancer and CRLM. M, Schematic of the CRLM mouse model. C57BL/6 mice received intrasplenic injections of mouse colorectal cancer cells (MC38). Mice were subsequently treated with IgG control (IgG; n = 6), anti-Ly6G antibody (anti-Ly6G; n = 6), or anti-Ly6G plus <t>anti-CD8a</t> antibodies (anti-Ly6G + anti-CD8a; n = 6). Livers were harvested on day 14. N, Representative images of liver metastases from IgG, anti-Ly6G, and anti-Ly6G + anti-CD8a groups. O, Comparison of liver weights among the three groups. P values were calculated by the Wilcoxon rank-sum test ( B and L ) and unpaired Student t test ( F , K , and O ). *, P < 0.05; **, P < 0.01; ***, P < 0.001. CRC, colorectal cancer; Ctrl, control.
    Anti Cd8a, supplied by Bio X Cell, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    cd8a  (Cytek Biosciences)
    91
    Cytek Biosciences cd8a
    Proangiogenic and immunosuppression phenotype of TANs in the CRLM microenvironment. A, Heatmap showing signature scores for angiogenesis, myeloid cell recruitment, neutrophil cytotoxicity, MDSC, and NET across each TAN and PMN subset. B, Comparison of angiogenesis, myeloid cell recruitment, MDSC, and NET signature scores among TAN and PMN subsets. C, Dot plot illustrating expression levels of major cytokines and chemokines in each TAN and PMN subset. D, Representative IHC images of CD31 staining in CRLM tissues. Scale bars, 50 µm. E, Correlation analysis between CD31 and TAN1 density using Spearman’s rank correlation ( n = 102). F, Representative images and quantification of tube formation in HUVECs when cocultured with conditioned medium from human LGALS3 − neutrophils or LGALS3 + neutrophils ( n = 6). G, UMAP plot showing six monocyte/macrophage subsets across colorectal cancer, adjacent liver tissues, CRLM, and PB samples. H, Shell-like plot depicting ligand–receptor interaction strength between TAN1_LGASL3 and neutrophil/monocyte/macrophage subset in CRLM. I, Selected ligand–receptor interactions between TAN1_LGASL3 and neutrophil/monocyte/macrophage subset in CRLM. J, Representative mIHC images of CD68, CCL3L1, CD66b, CCR1, and BHLHE40 in CRLM. Scale bars, 50 µm. K, Representative images of macrophage migration assays and effects of conditioned medium from human LGALS3 + neutrophils on migration of macrophages (induced from THP-1 cells) in the presence of CCR1 inhibitor (BX471) or anti-CCL3 antibody ( n = 6). L, Comparison of immunosuppressive macrophage signature scores between colorectal cancer and CRLM. M, Schematic of the CRLM mouse model. C57BL/6 mice received intrasplenic injections of mouse colorectal cancer cells (MC38). Mice were subsequently treated with IgG control (IgG; n = 6), anti-Ly6G antibody (anti-Ly6G; n = 6), or anti-Ly6G plus <t>anti-CD8a</t> antibodies (anti-Ly6G + anti-CD8a; n = 6). Livers were harvested on day 14. N, Representative images of liver metastases from IgG, anti-Ly6G, and anti-Ly6G + anti-CD8a groups. O, Comparison of liver weights among the three groups. P values were calculated by the Wilcoxon rank-sum test ( B and L ) and unpaired Student t test ( F , K , and O ). *, P < 0.05; **, P < 0.01; ***, P < 0.001. CRC, colorectal cancer; Ctrl, control.
    Cd8a, supplied by Cytek Biosciences, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    pe anti mouse cd8  (Proteintech)
    93
    Proteintech pe anti mouse cd8
    Proangiogenic and immunosuppression phenotype of TANs in the CRLM microenvironment. A, Heatmap showing signature scores for angiogenesis, myeloid cell recruitment, neutrophil cytotoxicity, MDSC, and NET across each TAN and PMN subset. B, Comparison of angiogenesis, myeloid cell recruitment, MDSC, and NET signature scores among TAN and PMN subsets. C, Dot plot illustrating expression levels of major cytokines and chemokines in each TAN and PMN subset. D, Representative IHC images of CD31 staining in CRLM tissues. Scale bars, 50 µm. E, Correlation analysis between CD31 and TAN1 density using Spearman’s rank correlation ( n = 102). F, Representative images and quantification of tube formation in HUVECs when cocultured with conditioned medium from human LGALS3 − neutrophils or LGALS3 + neutrophils ( n = 6). G, UMAP plot showing six monocyte/macrophage subsets across colorectal cancer, adjacent liver tissues, CRLM, and PB samples. H, Shell-like plot depicting ligand–receptor interaction strength between TAN1_LGASL3 and neutrophil/monocyte/macrophage subset in CRLM. I, Selected ligand–receptor interactions between TAN1_LGASL3 and neutrophil/monocyte/macrophage subset in CRLM. J, Representative mIHC images of CD68, CCL3L1, CD66b, CCR1, and BHLHE40 in CRLM. Scale bars, 50 µm. K, Representative images of macrophage migration assays and effects of conditioned medium from human LGALS3 + neutrophils on migration of macrophages (induced from THP-1 cells) in the presence of CCR1 inhibitor (BX471) or anti-CCL3 antibody ( n = 6). L, Comparison of immunosuppressive macrophage signature scores between colorectal cancer and CRLM. M, Schematic of the CRLM mouse model. C57BL/6 mice received intrasplenic injections of mouse colorectal cancer cells (MC38). Mice were subsequently treated with IgG control (IgG; n = 6), anti-Ly6G antibody (anti-Ly6G; n = 6), or anti-Ly6G plus <t>anti-CD8a</t> antibodies (anti-Ly6G + anti-CD8a; n = 6). Livers were harvested on day 14. N, Representative images of liver metastases from IgG, anti-Ly6G, and anti-Ly6G + anti-CD8a groups. O, Comparison of liver weights among the three groups. P values were calculated by the Wilcoxon rank-sum test ( B and L ) and unpaired Student t test ( F , K , and O ). *, P < 0.05; **, P < 0.01; ***, P < 0.001. CRC, colorectal cancer; Ctrl, control.
    Pe Anti Mouse Cd8, supplied by Proteintech, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/cd8a/PE+Anti-mouse+CD8a/pm42785880-55-14-21
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    pe anti mouse cd8 alpha antibody  (Elabscience Biotechnology)
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    Elabscience Biotechnology pe anti mouse cd8 alpha antibody
    Overexpression of UPF1 in ccRCC inhibits M2 macrophage polarization and enhances the tumor suppressor function of <t>CD8</t> + T cells, Caki-1 cell groups of sh-NC+PBS, sh-UPF1+PBS, or sh-UPF1 + 2-DG, and the conditioned medium was used to culture and polarize M0 macrophages. The polarized macrophages were cultured with activated CD8 + T cells. (A) CFSE assessed CD8 + T cell proliferative capacity. (B) Flow cytometry measured apoptosis rate of CD8 + T cells in different co-culture systems. (C) CD8 + T cells from culture systems were co-cultured with normal Caki-1 cells, with LDH assays determining cytotoxic effect of CD8 + T cells. *, p < 0.05. cRCC, Clear cell renal cell carcinoma; UPF1, Up-frameshift protein 1; 2-DG, 2-Deoxy-D-glucose; CFSE, Carboxyfluorescein succinimidyl ester.
    Pe Anti Mouse Cd8 Alpha Antibody, supplied by Elabscience Biotechnology, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    cd8 t cell isolation kit  (Miltenyi Biotec)
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    Miltenyi Biotec cd8 t cell isolation kit
    Overexpression of UPF1 in ccRCC inhibits M2 macrophage polarization and enhances the tumor suppressor function of <t>CD8</t> + T cells, Caki-1 cell groups of sh-NC+PBS, sh-UPF1+PBS, or sh-UPF1 + 2-DG, and the conditioned medium was used to culture and polarize M0 macrophages. The polarized macrophages were cultured with activated CD8 + T cells. (A) CFSE assessed CD8 + T cell proliferative capacity. (B) Flow cytometry measured apoptosis rate of CD8 + T cells in different co-culture systems. (C) CD8 + T cells from culture systems were co-cultured with normal Caki-1 cells, with LDH assays determining cytotoxic effect of CD8 + T cells. *, p < 0.05. cRCC, Clear cell renal cell carcinoma; UPF1, Up-frameshift protein 1; 2-DG, 2-Deoxy-D-glucose; CFSE, Carboxyfluorescein succinimidyl ester.
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    mouse cd8a t cell isolation kit  (Miltenyi Biotec)
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    Overexpression of UPF1 in ccRCC inhibits M2 macrophage polarization and enhances the tumor suppressor function of <t>CD8</t> + T cells, Caki-1 cell groups of sh-NC+PBS, sh-UPF1+PBS, or sh-UPF1 + 2-DG, and the conditioned medium was used to culture and polarize M0 macrophages. The polarized macrophages were cultured with activated CD8 + T cells. (A) CFSE assessed CD8 + T cell proliferative capacity. (B) Flow cytometry measured apoptosis rate of CD8 + T cells in different co-culture systems. (C) CD8 + T cells from culture systems were co-cultured with normal Caki-1 cells, with LDH assays determining cytotoxic effect of CD8 + T cells. *, p < 0.05. cRCC, Clear cell renal cell carcinoma; UPF1, Up-frameshift protein 1; 2-DG, 2-Deoxy-D-glucose; CFSE, Carboxyfluorescein succinimidyl ester.
    Mouse Cd8a T Cell Isolation Kit, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Proangiogenic and immunosuppression phenotype of TANs in the CRLM microenvironment. A, Heatmap showing signature scores for angiogenesis, myeloid cell recruitment, neutrophil cytotoxicity, MDSC, and NET across each TAN and PMN subset. B, Comparison of angiogenesis, myeloid cell recruitment, MDSC, and NET signature scores among TAN and PMN subsets. C, Dot plot illustrating expression levels of major cytokines and chemokines in each TAN and PMN subset. D, Representative IHC images of CD31 staining in CRLM tissues. Scale bars, 50 µm. E, Correlation analysis between CD31 and TAN1 density using Spearman’s rank correlation ( n = 102). F, Representative images and quantification of tube formation in HUVECs when cocultured with conditioned medium from human LGALS3 − neutrophils or LGALS3 + neutrophils ( n = 6). G, UMAP plot showing six monocyte/macrophage subsets across colorectal cancer, adjacent liver tissues, CRLM, and PB samples. H, Shell-like plot depicting ligand–receptor interaction strength between TAN1_LGASL3 and neutrophil/monocyte/macrophage subset in CRLM. I, Selected ligand–receptor interactions between TAN1_LGASL3 and neutrophil/monocyte/macrophage subset in CRLM. J, Representative mIHC images of CD68, CCL3L1, CD66b, CCR1, and BHLHE40 in CRLM. Scale bars, 50 µm. K, Representative images of macrophage migration assays and effects of conditioned medium from human LGALS3 + neutrophils on migration of macrophages (induced from THP-1 cells) in the presence of CCR1 inhibitor (BX471) or anti-CCL3 antibody ( n = 6). L, Comparison of immunosuppressive macrophage signature scores between colorectal cancer and CRLM. M, Schematic of the CRLM mouse model. C57BL/6 mice received intrasplenic injections of mouse colorectal cancer cells (MC38). Mice were subsequently treated with IgG control (IgG; n = 6), anti-Ly6G antibody (anti-Ly6G; n = 6), or anti-Ly6G plus anti-CD8a antibodies (anti-Ly6G + anti-CD8a; n = 6). Livers were harvested on day 14. N, Representative images of liver metastases from IgG, anti-Ly6G, and anti-Ly6G + anti-CD8a groups. O, Comparison of liver weights among the three groups. P values were calculated by the Wilcoxon rank-sum test ( B and L ) and unpaired Student t test ( F , K , and O ). *, P < 0.05; **, P < 0.01; ***, P < 0.001. CRC, colorectal cancer; Ctrl, control.

    Journal: Cancer Discovery

    Article Title: Senescent-Like Neutrophils Shape Angiogenic Immunosuppressive Niches in Colorectal Cancer Liver Metastasis

    doi: 10.1158/2159-8290.CD-25-0820

    Figure Lengend Snippet: Proangiogenic and immunosuppression phenotype of TANs in the CRLM microenvironment. A, Heatmap showing signature scores for angiogenesis, myeloid cell recruitment, neutrophil cytotoxicity, MDSC, and NET across each TAN and PMN subset. B, Comparison of angiogenesis, myeloid cell recruitment, MDSC, and NET signature scores among TAN and PMN subsets. C, Dot plot illustrating expression levels of major cytokines and chemokines in each TAN and PMN subset. D, Representative IHC images of CD31 staining in CRLM tissues. Scale bars, 50 µm. E, Correlation analysis between CD31 and TAN1 density using Spearman’s rank correlation ( n = 102). F, Representative images and quantification of tube formation in HUVECs when cocultured with conditioned medium from human LGALS3 − neutrophils or LGALS3 + neutrophils ( n = 6). G, UMAP plot showing six monocyte/macrophage subsets across colorectal cancer, adjacent liver tissues, CRLM, and PB samples. H, Shell-like plot depicting ligand–receptor interaction strength between TAN1_LGASL3 and neutrophil/monocyte/macrophage subset in CRLM. I, Selected ligand–receptor interactions between TAN1_LGASL3 and neutrophil/monocyte/macrophage subset in CRLM. J, Representative mIHC images of CD68, CCL3L1, CD66b, CCR1, and BHLHE40 in CRLM. Scale bars, 50 µm. K, Representative images of macrophage migration assays and effects of conditioned medium from human LGALS3 + neutrophils on migration of macrophages (induced from THP-1 cells) in the presence of CCR1 inhibitor (BX471) or anti-CCL3 antibody ( n = 6). L, Comparison of immunosuppressive macrophage signature scores between colorectal cancer and CRLM. M, Schematic of the CRLM mouse model. C57BL/6 mice received intrasplenic injections of mouse colorectal cancer cells (MC38). Mice were subsequently treated with IgG control (IgG; n = 6), anti-Ly6G antibody (anti-Ly6G; n = 6), or anti-Ly6G plus anti-CD8a antibodies (anti-Ly6G + anti-CD8a; n = 6). Livers were harvested on day 14. N, Representative images of liver metastases from IgG, anti-Ly6G, and anti-Ly6G + anti-CD8a groups. O, Comparison of liver weights among the three groups. P values were calculated by the Wilcoxon rank-sum test ( B and L ) and unpaired Student t test ( F , K , and O ). *, P < 0.05; **, P < 0.01; ***, P < 0.001. CRC, colorectal cancer; Ctrl, control.

    Article Snippet: For CD8 + T-cell depletion, 200 μg anti-mouse CD8a (Bio X Cell, BE0061, RRID: AB_1125541) was injected intraperitoneally twice a week.

    Techniques: Comparison, Expressing, Staining, Analysis, Migration, Control

    Overexpression of UPF1 in ccRCC inhibits M2 macrophage polarization and enhances the tumor suppressor function of CD8 + T cells, Caki-1 cell groups of sh-NC+PBS, sh-UPF1+PBS, or sh-UPF1 + 2-DG, and the conditioned medium was used to culture and polarize M0 macrophages. The polarized macrophages were cultured with activated CD8 + T cells. (A) CFSE assessed CD8 + T cell proliferative capacity. (B) Flow cytometry measured apoptosis rate of CD8 + T cells in different co-culture systems. (C) CD8 + T cells from culture systems were co-cultured with normal Caki-1 cells, with LDH assays determining cytotoxic effect of CD8 + T cells. *, p < 0.05. cRCC, Clear cell renal cell carcinoma; UPF1, Up-frameshift protein 1; 2-DG, 2-Deoxy-D-glucose; CFSE, Carboxyfluorescein succinimidyl ester.

    Journal: Frontiers in Immunology

    Article Title: UPF1-dependent glycolysis driven by LINC00887 facilitates macrophage M2 polarization in clear cell renal cell carcinoma

    doi: 10.3389/fimmu.2026.1833383

    Figure Lengend Snippet: Overexpression of UPF1 in ccRCC inhibits M2 macrophage polarization and enhances the tumor suppressor function of CD8 + T cells, Caki-1 cell groups of sh-NC+PBS, sh-UPF1+PBS, or sh-UPF1 + 2-DG, and the conditioned medium was used to culture and polarize M0 macrophages. The polarized macrophages were cultured with activated CD8 + T cells. (A) CFSE assessed CD8 + T cell proliferative capacity. (B) Flow cytometry measured apoptosis rate of CD8 + T cells in different co-culture systems. (C) CD8 + T cells from culture systems were co-cultured with normal Caki-1 cells, with LDH assays determining cytotoxic effect of CD8 + T cells. *, p < 0.05. cRCC, Clear cell renal cell carcinoma; UPF1, Up-frameshift protein 1; 2-DG, 2-Deoxy-D-glucose; CFSE, Carboxyfluorescein succinimidyl ester.

    Article Snippet: Antibodies included: FITC Mouse monoclonal to CD68 (ab134351, abcam, UK), Pacific Blue anti-human CD86 Antibody (305417, yajimall, China), CD206 (MMR) Monoclonal Antibody (19.2)-PE (12-2069-42, ThermoFisher, USA), PerCP/Cyanine5.5 Anti-Mouse F4/80 Antibody (E-AB-F0995J, Elabscience, China), PE Anti-CD206 mouse antibody (E-AB-F1135D, Elabscience, China), APC Anti-mouse CD3 antibody (E-AB-F1013E, Elabscience, China), PE Anti-mouse CD8 alpha antibody (E-AB-F1104D, Elabscience, China).

    Techniques: Over Expression, Cell Culture, Flow Cytometry, Co-culture Assay

    LINC00887/UPF1-mediated aerobic glycolysis leads to lactic acid accumulation and facilitates immune evasion in ccRCC in vivo , A syngeneic tumor model was developed by injecting BALB/c mice with Renca cells expressing sh-NC or sh-LINC00887, followed by treatment with sh-NC or sh-UPF1 lentivirus. A-C: Assessment of tumor size (A) , volume (B) , and weight (C) . (D, E) qRT-PCR analyses for LINC00887 (D) and UPF1 (E) mRNA levels. (F) IHC analysis for UPF1, LDH-A, KI67 protein levels. (G) Kit assay for lactate levels in tumors. (H) Proportion of M2 macrophages (F4/80+CD206+) by flow cytometry. (I) Immunofluorescence assay for CD206+ macrophages. (J) Proportion of CD8 + T cells (CD3 + CD8 + ) in tumor tissues. ns, no significant difference. * p < 0.05. cRCC, Clear cell renal cell carcinoma; UPF1, Up-frameshift protein 1; LDH-A, Lactate dehydrogenase A; CD206, Mannose receptor C-type 1; CD3, Cluster of differentiation 3; CD8, Cluster of differentiation 8.

    Journal: Frontiers in Immunology

    Article Title: UPF1-dependent glycolysis driven by LINC00887 facilitates macrophage M2 polarization in clear cell renal cell carcinoma

    doi: 10.3389/fimmu.2026.1833383

    Figure Lengend Snippet: LINC00887/UPF1-mediated aerobic glycolysis leads to lactic acid accumulation and facilitates immune evasion in ccRCC in vivo , A syngeneic tumor model was developed by injecting BALB/c mice with Renca cells expressing sh-NC or sh-LINC00887, followed by treatment with sh-NC or sh-UPF1 lentivirus. A-C: Assessment of tumor size (A) , volume (B) , and weight (C) . (D, E) qRT-PCR analyses for LINC00887 (D) and UPF1 (E) mRNA levels. (F) IHC analysis for UPF1, LDH-A, KI67 protein levels. (G) Kit assay for lactate levels in tumors. (H) Proportion of M2 macrophages (F4/80+CD206+) by flow cytometry. (I) Immunofluorescence assay for CD206+ macrophages. (J) Proportion of CD8 + T cells (CD3 + CD8 + ) in tumor tissues. ns, no significant difference. * p < 0.05. cRCC, Clear cell renal cell carcinoma; UPF1, Up-frameshift protein 1; LDH-A, Lactate dehydrogenase A; CD206, Mannose receptor C-type 1; CD3, Cluster of differentiation 3; CD8, Cluster of differentiation 8.

    Article Snippet: Antibodies included: FITC Mouse monoclonal to CD68 (ab134351, abcam, UK), Pacific Blue anti-human CD86 Antibody (305417, yajimall, China), CD206 (MMR) Monoclonal Antibody (19.2)-PE (12-2069-42, ThermoFisher, USA), PerCP/Cyanine5.5 Anti-Mouse F4/80 Antibody (E-AB-F0995J, Elabscience, China), PE Anti-CD206 mouse antibody (E-AB-F1135D, Elabscience, China), APC Anti-mouse CD3 antibody (E-AB-F1013E, Elabscience, China), PE Anti-mouse CD8 alpha antibody (E-AB-F1104D, Elabscience, China).

    Techniques: In Vivo, Expressing, Quantitative RT-PCR, Analysis, Flow Cytometry, Immunofluorescence