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microbead negative selection kits  (Miltenyi Biotec)


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    Miltenyi Biotec microbead negative selection kits
    Microbead Negative Selection Kits, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 97/100, based on 2782 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/negative+selection+kit/Pan+T+Cell+Isolation+Kit%2C+human/us12599650-561-11-18
    Average 97 stars, based on 2782 article reviews
    microbead negative selection kits - by Bioz Stars, 2026-10
    97/100 stars

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    Knockdown:

    Article Title: Close Contacts Unlocked: Nanopore-Stabilized Microvilli Bypass T Cell Receptor-Ligand-Dependent T Cell Activation.
    Article Snippet: For T cell isolation, the Naive Pan T cell Isolation kit (Miltenyi Biotec, #130−097−095, only α/β T cells were isolated) was utilized, following the manufacturer’s protocol. .. For TCR knockdown experiments or patient versus healthy pan T cell experiments, the Pan T cell Isolation kit (Miltenyi Biotec, #130−096−535) was used instead. ..

    Cell Isolation:

    Article Title: Close Contacts Unlocked: Nanopore-Stabilized Microvilli Bypass T Cell Receptor-Ligand-Dependent T Cell Activation.
    Article Snippet: For T cell isolation, the Naive Pan T cell Isolation kit (Miltenyi Biotec, #130−097−095, only α/β T cells were isolated) was utilized, following the manufacturer’s protocol. .. For TCR knockdown experiments or patient versus healthy pan T cell experiments, the Pan T cell Isolation kit (Miltenyi Biotec, #130−096−535) was used instead. ..

    Article Title: LC3-associated phagocytosis is impaired in monocyte-derived macrophages from systemic sclerosis patients
    Article Snippet: Accutase , – , Gibco, A1110501. .. Pan T cell Isolation Kit, human , – , Miltenyi Biotec, 130-096-535. .. CD14 MicroBeads, human , 20 μL/10 7 cells , Miltenyi Biotec, 130-050-201.

    Article Title: Single-allele nanoscale mapping of regulatory variants
    Article Snippet: .. The peripheral blood mononuclear cell layer was isolated and CD4 + or CD3 + pan-T cells were enriched with CD4 + magnetic beads (Miltenyi, cat. no. 130-045-101) or pan-T cell isolation kit (Miltenyi, cat. no. 130-096-535), respectively, as per the manufacturer’s instructions. .. CD4 + or CD3 + cells were cultured at 1 × 10 6 cells ml −1 in Immunocult-XF T cell expansion medium (STEMCELL Technologies, cat. no. 10981) supplemented with 25 μl ml −1 of Immunocult Human CD3, CD28, CD2 T cell activator (STEMCELL Technologies, cat. no. 100-0785) and 10 ng ml −1 of human interleukin-2 (Invitrogen), for 60 h, before collection.

    Article Title: Solid forms of pharmaceutically acceptable salts of (S)-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-4,6-dihydropyrrolo[3,4-d]imidazol-5(1H)-yl)(3-hydroxypyrrolidin-1-yl)methanone as JAK inhibitors
    Article Snippet: .. Experimental Scheme 2.1 Experimental Materials and Instruments Item Name Source/Supplier Catalogue No. 1 Human PBMC Cryopreserved TPCS PB050C 2 RPMI Medium 1640 Invitrogen A10491-01 3 Fetal bovine serum Invitrogen 10099-141 4 Penicillin-streptomycin Invitrogen 15140-122 5 DMSO MP 196055 6 Pan T Cell Isolation Kit, human Miltenyi Biotech 130-096-535 7 Recombinant Human IL-2 Protein R&D 202-IL-010 8 anti-CD3 human OKT3 eBioscience EB16-0037-85 9 CD28 Monoclonal Antibody eBioscience 16-0289-85 (CD28.2), Functional Grade 10 Tofacitinib Selleck S2789 11 Celltiter Glo assay kit Promega G7573 12 384 well plate Corning 3764 13 6 well plate Corning 3516 15 Plate Reader PerkinElmer Envision 2104 16 Centrifuge Eppendorf 5810R 17 Echo 550 PerkinElmer 6008280 2.2 Experimental Methods 2.2.1 Formulation of the Compound Stocks to be Tested Formulation of compound stocks Weighing Volume DMSO stock Molecular mass of DMSO concentration Compound weight (mg) added (μL) (mM) Tofacitinib 312.37 5.00 1600.70 10 Compound of 476.51 1.7 342.49 10 formula (I) 2.2.2 Experimental Steps of Kinase Reaction a) Isolating T cells from human PBMC using the Human Pan T Cell Isolation Kit according to the manual. b) Stimulating the T cells with anti-CD3 antibody and anti-CD28 antibody and incubating them for 72 h at 37° C. in a 5% CO2 incubator. c) Collecting T cells and washing the cells with PBS. d) Transferring 40 nL of the diluted compound solution to be tested to a 384-well plate using Echo 550. e) Inoculating the T cells (prepared in step c) into the 384-well reaction plate (prepared in step d) in an amount of 35 μL/well, centrifuging at 1000 rpm for 1 min, and incubating at 37° C. with 5% CO2. f) Adding 5 μL/well of human recombinant IL-2 protein at a final concentration of 10 ng/ml, then centrifuging at 1000 rpm for 1 min, and incubating for 72 h at 37° C. with 5% CO2. g) Adding 20 μL Celltiter Glo buffer per well, centrifuging at 1000 rpm for 1 min, mixing at 350 g for 2 min, and incubating at room temperature for 30 min. h) Reading the lumilencence signal values on the EnVision multifunctional plate reader. ..

    Article Title: Method for transduction of t cells in the presence of malignant cells
    Article Snippet: .. Primary T cells of 2 donors were isolated from PBMC using the PAN T cell isolation Kit (Miltenyi Biotec, 130-096-535) and cultivated in TexMACS medium (Miltenyi Biotec, 130-097-196) with IL7 (Miltenyi Biotec, 170-076-111) IL15 (Miltenyi Biotec, 170-076-114) and Transact (Miltenyi Biotec, 130-111-160) over night at 37° C. Half of the samples were treated with the integration inhibitor Raltegravir (Sigma Aldrich, cat. No. CDS023737-25MG). ..

    Isolation:

    Article Title: Single-allele nanoscale mapping of regulatory variants
    Article Snippet: .. The peripheral blood mononuclear cell layer was isolated and CD4 + or CD3 + pan-T cells were enriched with CD4 + magnetic beads (Miltenyi, cat. no. 130-045-101) or pan-T cell isolation kit (Miltenyi, cat. no. 130-096-535), respectively, as per the manufacturer’s instructions. .. CD4 + or CD3 + cells were cultured at 1 × 10 6 cells ml −1 in Immunocult-XF T cell expansion medium (STEMCELL Technologies, cat. no. 10981) supplemented with 25 μl ml −1 of Immunocult Human CD3, CD28, CD2 T cell activator (STEMCELL Technologies, cat. no. 100-0785) and 10 ng ml −1 of human interleukin-2 (Invitrogen), for 60 h, before collection.

    Article Title: Method for transduction of t cells in the presence of malignant cells
    Article Snippet: .. Primary T cells of 2 donors were isolated from PBMC using the PAN T cell isolation Kit (Miltenyi Biotec, 130-096-535) and cultivated in TexMACS medium (Miltenyi Biotec, 130-097-196) with IL7 (Miltenyi Biotec, 170-076-111) IL15 (Miltenyi Biotec, 170-076-114) and Transact (Miltenyi Biotec, 130-111-160) over night at 37° C. Half of the samples were treated with the integration inhibitor Raltegravir (Sigma Aldrich, cat. No. CDS023737-25MG). ..

    Magnetic Beads:

    Article Title: Single-allele nanoscale mapping of regulatory variants
    Article Snippet: .. The peripheral blood mononuclear cell layer was isolated and CD4 + or CD3 + pan-T cells were enriched with CD4 + magnetic beads (Miltenyi, cat. no. 130-045-101) or pan-T cell isolation kit (Miltenyi, cat. no. 130-096-535), respectively, as per the manufacturer’s instructions. .. CD4 + or CD3 + cells were cultured at 1 × 10 6 cells ml −1 in Immunocult-XF T cell expansion medium (STEMCELL Technologies, cat. no. 10981) supplemented with 25 μl ml −1 of Immunocult Human CD3, CD28, CD2 T cell activator (STEMCELL Technologies, cat. no. 100-0785) and 10 ng ml −1 of human interleukin-2 (Invitrogen), for 60 h, before collection.

    Recombinant:

    Article Title: Solid forms of pharmaceutically acceptable salts of (S)-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-4,6-dihydropyrrolo[3,4-d]imidazol-5(1H)-yl)(3-hydroxypyrrolidin-1-yl)methanone as JAK inhibitors
    Article Snippet: .. Experimental Scheme 2.1 Experimental Materials and Instruments Item Name Source/Supplier Catalogue No. 1 Human PBMC Cryopreserved TPCS PB050C 2 RPMI Medium 1640 Invitrogen A10491-01 3 Fetal bovine serum Invitrogen 10099-141 4 Penicillin-streptomycin Invitrogen 15140-122 5 DMSO MP 196055 6 Pan T Cell Isolation Kit, human Miltenyi Biotech 130-096-535 7 Recombinant Human IL-2 Protein R&D 202-IL-010 8 anti-CD3 human OKT3 eBioscience EB16-0037-85 9 CD28 Monoclonal Antibody eBioscience 16-0289-85 (CD28.2), Functional Grade 10 Tofacitinib Selleck S2789 11 Celltiter Glo assay kit Promega G7573 12 384 well plate Corning 3764 13 6 well plate Corning 3516 15 Plate Reader PerkinElmer Envision 2104 16 Centrifuge Eppendorf 5810R 17 Echo 550 PerkinElmer 6008280 2.2 Experimental Methods 2.2.1 Formulation of the Compound Stocks to be Tested Formulation of compound stocks Weighing Volume DMSO stock Molecular mass of DMSO concentration Compound weight (mg) added (μL) (mM) Tofacitinib 312.37 5.00 1600.70 10 Compound of 476.51 1.7 342.49 10 formula (I) 2.2.2 Experimental Steps of Kinase Reaction a) Isolating T cells from human PBMC using the Human Pan T Cell Isolation Kit according to the manual. b) Stimulating the T cells with anti-CD3 antibody and anti-CD28 antibody and incubating them for 72 h at 37° C. in a 5% CO2 incubator. c) Collecting T cells and washing the cells with PBS. d) Transferring 40 nL of the diluted compound solution to be tested to a 384-well plate using Echo 550. e) Inoculating the T cells (prepared in step c) into the 384-well reaction plate (prepared in step d) in an amount of 35 μL/well, centrifuging at 1000 rpm for 1 min, and incubating at 37° C. with 5% CO2. f) Adding 5 μL/well of human recombinant IL-2 protein at a final concentration of 10 ng/ml, then centrifuging at 1000 rpm for 1 min, and incubating for 72 h at 37° C. with 5% CO2. g) Adding 20 μL Celltiter Glo buffer per well, centrifuging at 1000 rpm for 1 min, mixing at 350 g for 2 min, and incubating at room temperature for 30 min. h) Reading the lumilencence signal values on the EnVision multifunctional plate reader. ..

    Functional Assay:

    Article Title: Solid forms of pharmaceutically acceptable salts of (S)-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-4,6-dihydropyrrolo[3,4-d]imidazol-5(1H)-yl)(3-hydroxypyrrolidin-1-yl)methanone as JAK inhibitors
    Article Snippet: .. Experimental Scheme 2.1 Experimental Materials and Instruments Item Name Source/Supplier Catalogue No. 1 Human PBMC Cryopreserved TPCS PB050C 2 RPMI Medium 1640 Invitrogen A10491-01 3 Fetal bovine serum Invitrogen 10099-141 4 Penicillin-streptomycin Invitrogen 15140-122 5 DMSO MP 196055 6 Pan T Cell Isolation Kit, human Miltenyi Biotech 130-096-535 7 Recombinant Human IL-2 Protein R&D 202-IL-010 8 anti-CD3 human OKT3 eBioscience EB16-0037-85 9 CD28 Monoclonal Antibody eBioscience 16-0289-85 (CD28.2), Functional Grade 10 Tofacitinib Selleck S2789 11 Celltiter Glo assay kit Promega G7573 12 384 well plate Corning 3764 13 6 well plate Corning 3516 15 Plate Reader PerkinElmer Envision 2104 16 Centrifuge Eppendorf 5810R 17 Echo 550 PerkinElmer 6008280 2.2 Experimental Methods 2.2.1 Formulation of the Compound Stocks to be Tested Formulation of compound stocks Weighing Volume DMSO stock Molecular mass of DMSO concentration Compound weight (mg) added (μL) (mM) Tofacitinib 312.37 5.00 1600.70 10 Compound of 476.51 1.7 342.49 10 formula (I) 2.2.2 Experimental Steps of Kinase Reaction a) Isolating T cells from human PBMC using the Human Pan T Cell Isolation Kit according to the manual. b) Stimulating the T cells with anti-CD3 antibody and anti-CD28 antibody and incubating them for 72 h at 37° C. in a 5% CO2 incubator. c) Collecting T cells and washing the cells with PBS. d) Transferring 40 nL of the diluted compound solution to be tested to a 384-well plate using Echo 550. e) Inoculating the T cells (prepared in step c) into the 384-well reaction plate (prepared in step d) in an amount of 35 μL/well, centrifuging at 1000 rpm for 1 min, and incubating at 37° C. with 5% CO2. f) Adding 5 μL/well of human recombinant IL-2 protein at a final concentration of 10 ng/ml, then centrifuging at 1000 rpm for 1 min, and incubating for 72 h at 37° C. with 5% CO2. g) Adding 20 μL Celltiter Glo buffer per well, centrifuging at 1000 rpm for 1 min, mixing at 350 g for 2 min, and incubating at room temperature for 30 min. h) Reading the lumilencence signal values on the EnVision multifunctional plate reader. ..

    Glo Assay:

    Article Title: Solid forms of pharmaceutically acceptable salts of (S)-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-4,6-dihydropyrrolo[3,4-d]imidazol-5(1H)-yl)(3-hydroxypyrrolidin-1-yl)methanone as JAK inhibitors
    Article Snippet: .. Experimental Scheme 2.1 Experimental Materials and Instruments Item Name Source/Supplier Catalogue No. 1 Human PBMC Cryopreserved TPCS PB050C 2 RPMI Medium 1640 Invitrogen A10491-01 3 Fetal bovine serum Invitrogen 10099-141 4 Penicillin-streptomycin Invitrogen 15140-122 5 DMSO MP 196055 6 Pan T Cell Isolation Kit, human Miltenyi Biotech 130-096-535 7 Recombinant Human IL-2 Protein R&D 202-IL-010 8 anti-CD3 human OKT3 eBioscience EB16-0037-85 9 CD28 Monoclonal Antibody eBioscience 16-0289-85 (CD28.2), Functional Grade 10 Tofacitinib Selleck S2789 11 Celltiter Glo assay kit Promega G7573 12 384 well plate Corning 3764 13 6 well plate Corning 3516 15 Plate Reader PerkinElmer Envision 2104 16 Centrifuge Eppendorf 5810R 17 Echo 550 PerkinElmer 6008280 2.2 Experimental Methods 2.2.1 Formulation of the Compound Stocks to be Tested Formulation of compound stocks Weighing Volume DMSO stock Molecular mass of DMSO concentration Compound weight (mg) added (μL) (mM) Tofacitinib 312.37 5.00 1600.70 10 Compound of 476.51 1.7 342.49 10 formula (I) 2.2.2 Experimental Steps of Kinase Reaction a) Isolating T cells from human PBMC using the Human Pan T Cell Isolation Kit according to the manual. b) Stimulating the T cells with anti-CD3 antibody and anti-CD28 antibody and incubating them for 72 h at 37° C. in a 5% CO2 incubator. c) Collecting T cells and washing the cells with PBS. d) Transferring 40 nL of the diluted compound solution to be tested to a 384-well plate using Echo 550. e) Inoculating the T cells (prepared in step c) into the 384-well reaction plate (prepared in step d) in an amount of 35 μL/well, centrifuging at 1000 rpm for 1 min, and incubating at 37° C. with 5% CO2. f) Adding 5 μL/well of human recombinant IL-2 protein at a final concentration of 10 ng/ml, then centrifuging at 1000 rpm for 1 min, and incubating for 72 h at 37° C. with 5% CO2. g) Adding 20 μL Celltiter Glo buffer per well, centrifuging at 1000 rpm for 1 min, mixing at 350 g for 2 min, and incubating at room temperature for 30 min. h) Reading the lumilencence signal values on the EnVision multifunctional plate reader. ..

    Formulation:

    Article Title: Solid forms of pharmaceutically acceptable salts of (S)-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-4,6-dihydropyrrolo[3,4-d]imidazol-5(1H)-yl)(3-hydroxypyrrolidin-1-yl)methanone as JAK inhibitors
    Article Snippet: .. Experimental Scheme 2.1 Experimental Materials and Instruments Item Name Source/Supplier Catalogue No. 1 Human PBMC Cryopreserved TPCS PB050C 2 RPMI Medium 1640 Invitrogen A10491-01 3 Fetal bovine serum Invitrogen 10099-141 4 Penicillin-streptomycin Invitrogen 15140-122 5 DMSO MP 196055 6 Pan T Cell Isolation Kit, human Miltenyi Biotech 130-096-535 7 Recombinant Human IL-2 Protein R&D 202-IL-010 8 anti-CD3 human OKT3 eBioscience EB16-0037-85 9 CD28 Monoclonal Antibody eBioscience 16-0289-85 (CD28.2), Functional Grade 10 Tofacitinib Selleck S2789 11 Celltiter Glo assay kit Promega G7573 12 384 well plate Corning 3764 13 6 well plate Corning 3516 15 Plate Reader PerkinElmer Envision 2104 16 Centrifuge Eppendorf 5810R 17 Echo 550 PerkinElmer 6008280 2.2 Experimental Methods 2.2.1 Formulation of the Compound Stocks to be Tested Formulation of compound stocks Weighing Volume DMSO stock Molecular mass of DMSO concentration Compound weight (mg) added (μL) (mM) Tofacitinib 312.37 5.00 1600.70 10 Compound of 476.51 1.7 342.49 10 formula (I) 2.2.2 Experimental Steps of Kinase Reaction a) Isolating T cells from human PBMC using the Human Pan T Cell Isolation Kit according to the manual. b) Stimulating the T cells with anti-CD3 antibody and anti-CD28 antibody and incubating them for 72 h at 37° C. in a 5% CO2 incubator. c) Collecting T cells and washing the cells with PBS. d) Transferring 40 nL of the diluted compound solution to be tested to a 384-well plate using Echo 550. e) Inoculating the T cells (prepared in step c) into the 384-well reaction plate (prepared in step d) in an amount of 35 μL/well, centrifuging at 1000 rpm for 1 min, and incubating at 37° C. with 5% CO2. f) Adding 5 μL/well of human recombinant IL-2 protein at a final concentration of 10 ng/ml, then centrifuging at 1000 rpm for 1 min, and incubating for 72 h at 37° C. with 5% CO2. g) Adding 20 μL Celltiter Glo buffer per well, centrifuging at 1000 rpm for 1 min, mixing at 350 g for 2 min, and incubating at room temperature for 30 min. h) Reading the lumilencence signal values on the EnVision multifunctional plate reader. ..

    Concentration Assay:

    Article Title: Solid forms of pharmaceutically acceptable salts of (S)-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-4,6-dihydropyrrolo[3,4-d]imidazol-5(1H)-yl)(3-hydroxypyrrolidin-1-yl)methanone as JAK inhibitors
    Article Snippet: .. Experimental Scheme 2.1 Experimental Materials and Instruments Item Name Source/Supplier Catalogue No. 1 Human PBMC Cryopreserved TPCS PB050C 2 RPMI Medium 1640 Invitrogen A10491-01 3 Fetal bovine serum Invitrogen 10099-141 4 Penicillin-streptomycin Invitrogen 15140-122 5 DMSO MP 196055 6 Pan T Cell Isolation Kit, human Miltenyi Biotech 130-096-535 7 Recombinant Human IL-2 Protein R&D 202-IL-010 8 anti-CD3 human OKT3 eBioscience EB16-0037-85 9 CD28 Monoclonal Antibody eBioscience 16-0289-85 (CD28.2), Functional Grade 10 Tofacitinib Selleck S2789 11 Celltiter Glo assay kit Promega G7573 12 384 well plate Corning 3764 13 6 well plate Corning 3516 15 Plate Reader PerkinElmer Envision 2104 16 Centrifuge Eppendorf 5810R 17 Echo 550 PerkinElmer 6008280 2.2 Experimental Methods 2.2.1 Formulation of the Compound Stocks to be Tested Formulation of compound stocks Weighing Volume DMSO stock Molecular mass of DMSO concentration Compound weight (mg) added (μL) (mM) Tofacitinib 312.37 5.00 1600.70 10 Compound of 476.51 1.7 342.49 10 formula (I) 2.2.2 Experimental Steps of Kinase Reaction a) Isolating T cells from human PBMC using the Human Pan T Cell Isolation Kit according to the manual. b) Stimulating the T cells with anti-CD3 antibody and anti-CD28 antibody and incubating them for 72 h at 37° C. in a 5% CO2 incubator. c) Collecting T cells and washing the cells with PBS. d) Transferring 40 nL of the diluted compound solution to be tested to a 384-well plate using Echo 550. e) Inoculating the T cells (prepared in step c) into the 384-well reaction plate (prepared in step d) in an amount of 35 μL/well, centrifuging at 1000 rpm for 1 min, and incubating at 37° C. with 5% CO2. f) Adding 5 μL/well of human recombinant IL-2 protein at a final concentration of 10 ng/ml, then centrifuging at 1000 rpm for 1 min, and incubating for 72 h at 37° C. with 5% CO2. g) Adding 20 μL Celltiter Glo buffer per well, centrifuging at 1000 rpm for 1 min, mixing at 350 g for 2 min, and incubating at room temperature for 30 min. h) Reading the lumilencence signal values on the EnVision multifunctional plate reader. ..

    Transferring:

    Article Title: Solid forms of pharmaceutically acceptable salts of (S)-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-4,6-dihydropyrrolo[3,4-d]imidazol-5(1H)-yl)(3-hydroxypyrrolidin-1-yl)methanone as JAK inhibitors
    Article Snippet: .. Experimental Scheme 2.1 Experimental Materials and Instruments Item Name Source/Supplier Catalogue No. 1 Human PBMC Cryopreserved TPCS PB050C 2 RPMI Medium 1640 Invitrogen A10491-01 3 Fetal bovine serum Invitrogen 10099-141 4 Penicillin-streptomycin Invitrogen 15140-122 5 DMSO MP 196055 6 Pan T Cell Isolation Kit, human Miltenyi Biotech 130-096-535 7 Recombinant Human IL-2 Protein R&D 202-IL-010 8 anti-CD3 human OKT3 eBioscience EB16-0037-85 9 CD28 Monoclonal Antibody eBioscience 16-0289-85 (CD28.2), Functional Grade 10 Tofacitinib Selleck S2789 11 Celltiter Glo assay kit Promega G7573 12 384 well plate Corning 3764 13 6 well plate Corning 3516 15 Plate Reader PerkinElmer Envision 2104 16 Centrifuge Eppendorf 5810R 17 Echo 550 PerkinElmer 6008280 2.2 Experimental Methods 2.2.1 Formulation of the Compound Stocks to be Tested Formulation of compound stocks Weighing Volume DMSO stock Molecular mass of DMSO concentration Compound weight (mg) added (μL) (mM) Tofacitinib 312.37 5.00 1600.70 10 Compound of 476.51 1.7 342.49 10 formula (I) 2.2.2 Experimental Steps of Kinase Reaction a) Isolating T cells from human PBMC using the Human Pan T Cell Isolation Kit according to the manual. b) Stimulating the T cells with anti-CD3 antibody and anti-CD28 antibody and incubating them for 72 h at 37° C. in a 5% CO2 incubator. c) Collecting T cells and washing the cells with PBS. d) Transferring 40 nL of the diluted compound solution to be tested to a 384-well plate using Echo 550. e) Inoculating the T cells (prepared in step c) into the 384-well reaction plate (prepared in step d) in an amount of 35 μL/well, centrifuging at 1000 rpm for 1 min, and incubating at 37° C. with 5% CO2. f) Adding 5 μL/well of human recombinant IL-2 protein at a final concentration of 10 ng/ml, then centrifuging at 1000 rpm for 1 min, and incubating for 72 h at 37° C. with 5% CO2. g) Adding 20 μL Celltiter Glo buffer per well, centrifuging at 1000 rpm for 1 min, mixing at 350 g for 2 min, and incubating at room temperature for 30 min. h) Reading the lumilencence signal values on the EnVision multifunctional plate reader. ..



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    mBH3@NPs induced effective antitumor immunity and remodeled the tumor microenvironment (A) Experimental timeline for mBH3@NPs administration in B16-F10-tumor-bearing mice. Mice were administered with mBH3@NPs three times, with a dosing interval of every 2 days (B) Average tumor volume curves for mice treated in the melanoma model ( n = 8). (C) Individual tumor volume curves for mice treated in the melanoma model ( n = 8). (D) Western blot analysis of Bax, Bak, Bcl-2, Bcl-x L , Mcl-1, Puma, Bim, caspase-3, and caspase-9 expression following different treatments. β-Actin was used as a loading control ( n = 3). (E) Representative images of tumor immunostaining and quantification for TUNEL (green)/DAPI (blue), CD8 (red)/DAPI (blue), and CRT (green)/DAPI (blue) at day 19 following the indicated treatments in the melanoma model. Scale bars, 200 μm. (F) HMGB1 expression in tumor tissues analyzed by ELISA following different treatments ( n = 5). (G) Heatmap of cytokine expression levels (GrB, IFN-γ, TNF-α, IL-12, and IL-6) in tumor tissues following different treatments ( n = 5). (H) Flow cytometric analysis of immune cell populations (T cells, CD8 + T cells, CD4 + T cells, Treg cells, NK cells, macrophages, M1-like TAMs, M2-like TAMs, and MDSCs) within tumors and CD11c + CD80 + CD86 + mature dendritic cells in lymph nodes ( n = 5). One-way ANOVA with Tukey’s multiple comparisons test was used for all statistical analyses. Data are presented as the mean ± SD. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; NS, not significant. See also and .
    Negative Selection 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
    https://www.bioz.com/product/negative+selection+kit/NK+Cell+Isolation+Kit%2C+human/pm42013130-259-8-11
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    human cd8 t cell negative selection kit  (MedChemExpress)
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    mBH3@NPs induced effective antitumor immunity and remodeled the tumor microenvironment (A) Experimental timeline for mBH3@NPs administration in B16-F10-tumor-bearing mice. Mice were administered with mBH3@NPs three times, with a dosing interval of every 2 days (B) Average tumor volume curves for mice treated in the melanoma model ( n = 8). (C) Individual tumor volume curves for mice treated in the melanoma model ( n = 8). (D) Western blot analysis of Bax, Bak, Bcl-2, Bcl-x L , Mcl-1, Puma, Bim, caspase-3, and caspase-9 expression following different treatments. β-Actin was used as a loading control ( n = 3). (E) Representative images of tumor immunostaining and quantification for TUNEL (green)/DAPI (blue), <t>CD8</t> (red)/DAPI (blue), and CRT (green)/DAPI (blue) at day 19 following the indicated treatments in the melanoma model. Scale bars, 200 μm. (F) HMGB1 expression in tumor tissues analyzed by ELISA following different treatments ( n = 5). (G) Heatmap of cytokine expression levels (GrB, IFN-γ, TNF-α, IL-12, and IL-6) in tumor tissues following different treatments ( n = 5). (H) Flow cytometric analysis of immune cell populations (T cells, CD8 + T cells, CD4 + T cells, Treg cells, NK cells, macrophages, M1-like TAMs, M2-like TAMs, and MDSCs) within tumors and CD11c + CD80 + CD86 + mature dendritic cells in lymph nodes ( n = 5). One-way ANOVA with Tukey’s multiple comparisons test was used for all statistical analyses. Data are presented as the mean ± SD. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; NS, not significant. See also and .
    Human Cd8 T Cell Negative Selection Kit, supplied by MedChemExpress, 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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    mBH3@NPs induced effective antitumor immunity and remodeled the tumor microenvironment (A) Experimental timeline for mBH3@NPs administration in B16-F10-tumor-bearing mice. Mice were administered with mBH3@NPs three times, with a dosing interval of every 2 days (B) Average tumor volume curves for mice treated in the melanoma model ( n = 8). (C) Individual tumor volume curves for mice treated in the melanoma model ( n = 8). (D) Western blot analysis of Bax, Bak, Bcl-2, Bcl-x L , Mcl-1, Puma, Bim, caspase-3, and caspase-9 expression following different treatments. β-Actin was used as a loading control ( n = 3). (E) Representative images of tumor immunostaining and quantification for TUNEL (green)/DAPI (blue), <t>CD8</t> (red)/DAPI (blue), and CRT (green)/DAPI (blue) at day 19 following the indicated treatments in the melanoma model. Scale bars, 200 μm. (F) HMGB1 expression in tumor tissues analyzed by ELISA following different treatments ( n = 5). (G) Heatmap of cytokine expression levels (GrB, IFN-γ, TNF-α, IL-12, and IL-6) in tumor tissues following different treatments ( n = 5). (H) Flow cytometric analysis of immune cell populations (T cells, CD8 + T cells, CD4 + T cells, Treg cells, NK cells, macrophages, M1-like TAMs, M2-like TAMs, and MDSCs) within tumors and CD11c + CD80 + CD86 + mature dendritic cells in lymph nodes ( n = 5). One-way ANOVA with Tukey’s multiple comparisons test was used for all statistical analyses. Data are presented as the mean ± SD. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; NS, not significant. See also and .
    Negative Selection Macs Separation Kit, 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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    Image Search Results


    mBH3@NPs induced effective antitumor immunity and remodeled the tumor microenvironment (A) Experimental timeline for mBH3@NPs administration in B16-F10-tumor-bearing mice. Mice were administered with mBH3@NPs three times, with a dosing interval of every 2 days (B) Average tumor volume curves for mice treated in the melanoma model ( n = 8). (C) Individual tumor volume curves for mice treated in the melanoma model ( n = 8). (D) Western blot analysis of Bax, Bak, Bcl-2, Bcl-x L , Mcl-1, Puma, Bim, caspase-3, and caspase-9 expression following different treatments. β-Actin was used as a loading control ( n = 3). (E) Representative images of tumor immunostaining and quantification for TUNEL (green)/DAPI (blue), CD8 (red)/DAPI (blue), and CRT (green)/DAPI (blue) at day 19 following the indicated treatments in the melanoma model. Scale bars, 200 μm. (F) HMGB1 expression in tumor tissues analyzed by ELISA following different treatments ( n = 5). (G) Heatmap of cytokine expression levels (GrB, IFN-γ, TNF-α, IL-12, and IL-6) in tumor tissues following different treatments ( n = 5). (H) Flow cytometric analysis of immune cell populations (T cells, CD8 + T cells, CD4 + T cells, Treg cells, NK cells, macrophages, M1-like TAMs, M2-like TAMs, and MDSCs) within tumors and CD11c + CD80 + CD86 + mature dendritic cells in lymph nodes ( n = 5). One-way ANOVA with Tukey’s multiple comparisons test was used for all statistical analyses. Data are presented as the mean ± SD. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; NS, not significant. See also and .

    Journal: Cell Reports Medicine

    Article Title: mRNA lipid-nanoparticle-mediated mitochondrial apoptosis augments adoptive T cell immunotherapy

    doi: 10.1016/j.xcrm.2026.102706

    Figure Lengend Snippet: mBH3@NPs induced effective antitumor immunity and remodeled the tumor microenvironment (A) Experimental timeline for mBH3@NPs administration in B16-F10-tumor-bearing mice. Mice were administered with mBH3@NPs three times, with a dosing interval of every 2 days (B) Average tumor volume curves for mice treated in the melanoma model ( n = 8). (C) Individual tumor volume curves for mice treated in the melanoma model ( n = 8). (D) Western blot analysis of Bax, Bak, Bcl-2, Bcl-x L , Mcl-1, Puma, Bim, caspase-3, and caspase-9 expression following different treatments. β-Actin was used as a loading control ( n = 3). (E) Representative images of tumor immunostaining and quantification for TUNEL (green)/DAPI (blue), CD8 (red)/DAPI (blue), and CRT (green)/DAPI (blue) at day 19 following the indicated treatments in the melanoma model. Scale bars, 200 μm. (F) HMGB1 expression in tumor tissues analyzed by ELISA following different treatments ( n = 5). (G) Heatmap of cytokine expression levels (GrB, IFN-γ, TNF-α, IL-12, and IL-6) in tumor tissues following different treatments ( n = 5). (H) Flow cytometric analysis of immune cell populations (T cells, CD8 + T cells, CD4 + T cells, Treg cells, NK cells, macrophages, M1-like TAMs, M2-like TAMs, and MDSCs) within tumors and CD11c + CD80 + CD86 + mature dendritic cells in lymph nodes ( n = 5). One-way ANOVA with Tukey’s multiple comparisons test was used for all statistical analyses. Data are presented as the mean ± SD. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; NS, not significant. See also and .

    Article Snippet: Human CD8 + T cell Negative Selection , MedChemExpress , Cat#HY-KO351.

    Techniques: Western Blot, Expressing, Control, Immunostaining, TUNEL Assay, Enzyme-linked Immunosorbent Assay

    mBH3@NPs promoted the recruitment of adoptively transferred T cells (A) Schematic of transwell migration assay using B16-OVA cells and preactivated OT-1 T cells. (B) Transwell migration assay of OT-I T cells (CFDA-labeled). Representative flow cytometry graphs and quantification of T cell infiltration relative fold changes (calculated as the ratio of bottom well-located OT-I T cells to counting beads) ( n = 3). (C) Experimental timeline for mBH3@NPs administration combined with adoptively transferred T cells in B16-OVA tumor-bearing mice. (D) Serial IVIS images and quantification of infiltrating Cy5-T cell signals in B16-OVA tumor-bearing mice ( n = 5). (E) Quantification of tumor-infiltrating endogenous T cells and transferred CD8 + T cells per million cells ( n = 5). (F) Heatmap of RNA-seq data for key interleukin family member, chemokine family member, and granzyme family member from harvested tumor tissues, plotted as Z score of normalized gene expression for each gene ( n = 3). (G) Heatmap of the differences in pathway activities scored by GSVA from harvested tumor tissues, plotted as Z score of normalized gene expression for each gene ( n = 3). (H) Validation of key cytokines and chemokines expression by multiplex cytokine assay. Tumor tissue was homogenized, and the cell supernatant of each treatment group was collected ( n = 4). One-way ANOVA with Tukey’s multiple comparisons test was used for all statistical analyses. Data are presented as the mean ± SD. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; NS, not significant. See also .

    Journal: Cell Reports Medicine

    Article Title: mRNA lipid-nanoparticle-mediated mitochondrial apoptosis augments adoptive T cell immunotherapy

    doi: 10.1016/j.xcrm.2026.102706

    Figure Lengend Snippet: mBH3@NPs promoted the recruitment of adoptively transferred T cells (A) Schematic of transwell migration assay using B16-OVA cells and preactivated OT-1 T cells. (B) Transwell migration assay of OT-I T cells (CFDA-labeled). Representative flow cytometry graphs and quantification of T cell infiltration relative fold changes (calculated as the ratio of bottom well-located OT-I T cells to counting beads) ( n = 3). (C) Experimental timeline for mBH3@NPs administration combined with adoptively transferred T cells in B16-OVA tumor-bearing mice. (D) Serial IVIS images and quantification of infiltrating Cy5-T cell signals in B16-OVA tumor-bearing mice ( n = 5). (E) Quantification of tumor-infiltrating endogenous T cells and transferred CD8 + T cells per million cells ( n = 5). (F) Heatmap of RNA-seq data for key interleukin family member, chemokine family member, and granzyme family member from harvested tumor tissues, plotted as Z score of normalized gene expression for each gene ( n = 3). (G) Heatmap of the differences in pathway activities scored by GSVA from harvested tumor tissues, plotted as Z score of normalized gene expression for each gene ( n = 3). (H) Validation of key cytokines and chemokines expression by multiplex cytokine assay. Tumor tissue was homogenized, and the cell supernatant of each treatment group was collected ( n = 4). One-way ANOVA with Tukey’s multiple comparisons test was used for all statistical analyses. Data are presented as the mean ± SD. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; NS, not significant. See also .

    Article Snippet: Human CD8 + T cell Negative Selection , MedChemExpress , Cat#HY-KO351.

    Techniques: Transwell Migration Assay, Labeling, Flow Cytometry, RNA Sequencing, Gene Expression, Biomarker Discovery, Expressing, Multiplex Assay, Cytokine Assay

    mBH3@NPs combined with adoptive T cell therapy improved the therapeutic effect (A) Experimental timeline for mBH3@NPs administration combined with adoptively transferred T cells from PMEL mice in B16-F10-tumor-bearing mice. (B) The average tumor volume curves for mice treated in the B16-F10-tumor-bearing mice model ( n = 7). (C) Survival analysis for mice treated in B16-F10-tumor-bearing mice model ( n = 7). Mice were humanely euthanized when tumor volume reached the predefined ethical endpoint (15 mm × 15 mm). The x axis indicates days post-mBH3@NPs administration. (D) Experimental timeline for mBH3@NPs administration combined with adoptive transfered T cells from OT-1 mice in B16-OVA-tumor-bearing mice. (E) The average tumor volume curves for mice treated in B16-OVA-tumor-bearing mice model ( n = 7). (F) Survival analysis for mice treated in B16-OVA-tumor-bearing mice model ( n = 7). Mice were humanely euthanized when tumor volume reached the predefined ethical endpoint (15 mm × 15 mm). (G) Representative flow cytometry plot and quantification of CD44 and CD62L expression levels gated on total CD8 + T cells (upper panel) and the percentage of endogenous (CD90.2) CD8 + T cells and transferred PMEL (CD90.1) CD8 + T cells gated on total CD8 + T cells (lower panel) ( n = 5). (H) Representative flow cytometry plot and quantification of the expression level of granzyme B (GrB) and perforin in total CD8 + T cells from tumors and spleens ( n = 5). (I) Representative flow cytometry plot and quantification of the expression level of PD-1 and LAG-3 in both endogenous and transferred PMEL CD8 + T cells from tumors ( n = 5). (J) Immunofluorescence evaluation of CD8 + T cell exhaustion, effector, and memory function. Upper panel: representative immunofluorescence images of tumors from different groups for analyzing the exhaustion (PD-1 + , cyan) and effector function (IFN-γ + , green) of CD8 + T cells (CD8 + , pink). Scale bars, 100 μm. Lower panel: representative immunofluorescence images of tumor-infiltrating CD8 + T cells (CD8 + , green) along with markers for activation (CD69 + , red) and memory differentiation (CD44 + , pink; CCR7 + , cyan). Scale bars, 200 μm ( n = 3). One-way ANOVA with Tukey’s multiple comparisons test was used for all statistical analyses. Data are presented as the mean ± SD. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; NS, not significant. See also .

    Journal: Cell Reports Medicine

    Article Title: mRNA lipid-nanoparticle-mediated mitochondrial apoptosis augments adoptive T cell immunotherapy

    doi: 10.1016/j.xcrm.2026.102706

    Figure Lengend Snippet: mBH3@NPs combined with adoptive T cell therapy improved the therapeutic effect (A) Experimental timeline for mBH3@NPs administration combined with adoptively transferred T cells from PMEL mice in B16-F10-tumor-bearing mice. (B) The average tumor volume curves for mice treated in the B16-F10-tumor-bearing mice model ( n = 7). (C) Survival analysis for mice treated in B16-F10-tumor-bearing mice model ( n = 7). Mice were humanely euthanized when tumor volume reached the predefined ethical endpoint (15 mm × 15 mm). The x axis indicates days post-mBH3@NPs administration. (D) Experimental timeline for mBH3@NPs administration combined with adoptive transfered T cells from OT-1 mice in B16-OVA-tumor-bearing mice. (E) The average tumor volume curves for mice treated in B16-OVA-tumor-bearing mice model ( n = 7). (F) Survival analysis for mice treated in B16-OVA-tumor-bearing mice model ( n = 7). Mice were humanely euthanized when tumor volume reached the predefined ethical endpoint (15 mm × 15 mm). (G) Representative flow cytometry plot and quantification of CD44 and CD62L expression levels gated on total CD8 + T cells (upper panel) and the percentage of endogenous (CD90.2) CD8 + T cells and transferred PMEL (CD90.1) CD8 + T cells gated on total CD8 + T cells (lower panel) ( n = 5). (H) Representative flow cytometry plot and quantification of the expression level of granzyme B (GrB) and perforin in total CD8 + T cells from tumors and spleens ( n = 5). (I) Representative flow cytometry plot and quantification of the expression level of PD-1 and LAG-3 in both endogenous and transferred PMEL CD8 + T cells from tumors ( n = 5). (J) Immunofluorescence evaluation of CD8 + T cell exhaustion, effector, and memory function. Upper panel: representative immunofluorescence images of tumors from different groups for analyzing the exhaustion (PD-1 + , cyan) and effector function (IFN-γ + , green) of CD8 + T cells (CD8 + , pink). Scale bars, 100 μm. Lower panel: representative immunofluorescence images of tumor-infiltrating CD8 + T cells (CD8 + , green) along with markers for activation (CD69 + , red) and memory differentiation (CD44 + , pink; CCR7 + , cyan). Scale bars, 200 μm ( n = 3). One-way ANOVA with Tukey’s multiple comparisons test was used for all statistical analyses. Data are presented as the mean ± SD. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; NS, not significant. See also .

    Article Snippet: Human CD8 + T cell Negative Selection , MedChemExpress , Cat#HY-KO351.

    Techniques: Flow Cytometry, Expressing, Immunofluorescence, Activation Assay

    Combination therapy drove CD8 + T cells toward memory-like states with expanded TCR diversity (A) Experimental design for scRNA-seq. Mice were treated with ACT alone or combined with mBim@NPs and mPuma@NPs as shown ( n = 3); CD3 + T cell isolated from mice for scRNA-seq. (B) UMAP representation and distribution of 68,176 cells of the CD8 + T cell atlas in each experimental condition, colored by 12 CD8 + T cell subtypes annotated in this study. (C) UMAP representation of the CD8 + T cells atlas colored by each experimental condition. (D) A dot plot showing the expression levels of marker genes in each CD8 + T cell subtype. The color of the dots indicates the average scaled expression level, and the size of the dots indicates the percentage of cells expressing the gene in each subtype. (E) Indicated expression of gene marker associated with memory ( Tcf7 , Il7r , Sell , and Ccl5 ), exhaustion ( Pdcd1 , Tigit , and Lag3 ), and cytotoxicity ( Gzmb , Ifng , and Prf1 ) in the UMAP plot. (F) A dot plot showing the expression levels of marker genes in E. The color of the dots indicates the average scaled expression level, and the size of the dots indicates the percentage of cells expressing the gene in each subtype. (G) Volcano plots of log 2 (fold change) and log 10 (adjusted FDR value) of differentially expressed genes in proliferating T and Gzmk + T rm from mice treated with ACT monotherapy and combination therapy with mBim@NPs. (H) Shannon entropy of the T ex , proliferating T, T naive , and Gzmk + T rm subsets across different treatment groups (T: ACT monotherapy; TP: ACT + mPuma@NPs; TB: ACT + mBim@NPs). (I) Shannon entropy of total CD8 + T cells and CD4 + T cells across different treatment groups. (J) A heatmap of normalized Shannon entropy for CD8 + T cell subsets across different treatment groups. See also .

    Journal: Cell Reports Medicine

    Article Title: mRNA lipid-nanoparticle-mediated mitochondrial apoptosis augments adoptive T cell immunotherapy

    doi: 10.1016/j.xcrm.2026.102706

    Figure Lengend Snippet: Combination therapy drove CD8 + T cells toward memory-like states with expanded TCR diversity (A) Experimental design for scRNA-seq. Mice were treated with ACT alone or combined with mBim@NPs and mPuma@NPs as shown ( n = 3); CD3 + T cell isolated from mice for scRNA-seq. (B) UMAP representation and distribution of 68,176 cells of the CD8 + T cell atlas in each experimental condition, colored by 12 CD8 + T cell subtypes annotated in this study. (C) UMAP representation of the CD8 + T cells atlas colored by each experimental condition. (D) A dot plot showing the expression levels of marker genes in each CD8 + T cell subtype. The color of the dots indicates the average scaled expression level, and the size of the dots indicates the percentage of cells expressing the gene in each subtype. (E) Indicated expression of gene marker associated with memory ( Tcf7 , Il7r , Sell , and Ccl5 ), exhaustion ( Pdcd1 , Tigit , and Lag3 ), and cytotoxicity ( Gzmb , Ifng , and Prf1 ) in the UMAP plot. (F) A dot plot showing the expression levels of marker genes in E. The color of the dots indicates the average scaled expression level, and the size of the dots indicates the percentage of cells expressing the gene in each subtype. (G) Volcano plots of log 2 (fold change) and log 10 (adjusted FDR value) of differentially expressed genes in proliferating T and Gzmk + T rm from mice treated with ACT monotherapy and combination therapy with mBim@NPs. (H) Shannon entropy of the T ex , proliferating T, T naive , and Gzmk + T rm subsets across different treatment groups (T: ACT monotherapy; TP: ACT + mPuma@NPs; TB: ACT + mBim@NPs). (I) Shannon entropy of total CD8 + T cells and CD4 + T cells across different treatment groups. (J) A heatmap of normalized Shannon entropy for CD8 + T cell subsets across different treatment groups. See also .

    Article Snippet: Human CD8 + T cell Negative Selection , MedChemExpress , Cat#HY-KO351.

    Techniques: Isolation, Expressing, Marker

    mBH3@NPs induced effective antitumor immunity and remodeled the tumor microenvironment (A) Experimental timeline for mBH3@NPs administration in B16-F10-tumor-bearing mice. Mice were administered with mBH3@NPs three times, with a dosing interval of every 2 days (B) Average tumor volume curves for mice treated in the melanoma model ( n = 8). (C) Individual tumor volume curves for mice treated in the melanoma model ( n = 8). (D) Western blot analysis of Bax, Bak, Bcl-2, Bcl-x L , Mcl-1, Puma, Bim, caspase-3, and caspase-9 expression following different treatments. β-Actin was used as a loading control ( n = 3). (E) Representative images of tumor immunostaining and quantification for TUNEL (green)/DAPI (blue), CD8 (red)/DAPI (blue), and CRT (green)/DAPI (blue) at day 19 following the indicated treatments in the melanoma model. Scale bars, 200 μm. (F) HMGB1 expression in tumor tissues analyzed by ELISA following different treatments ( n = 5). (G) Heatmap of cytokine expression levels (GrB, IFN-γ, TNF-α, IL-12, and IL-6) in tumor tissues following different treatments ( n = 5). (H) Flow cytometric analysis of immune cell populations (T cells, CD8 + T cells, CD4 + T cells, Treg cells, NK cells, macrophages, M1-like TAMs, M2-like TAMs, and MDSCs) within tumors and CD11c + CD80 + CD86 + mature dendritic cells in lymph nodes ( n = 5). One-way ANOVA with Tukey’s multiple comparisons test was used for all statistical analyses. Data are presented as the mean ± SD. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; NS, not significant. See also and .

    Journal: Cell Reports Medicine

    Article Title: mRNA lipid-nanoparticle-mediated mitochondrial apoptosis augments adoptive T cell immunotherapy

    doi: 10.1016/j.xcrm.2026.102706

    Figure Lengend Snippet: mBH3@NPs induced effective antitumor immunity and remodeled the tumor microenvironment (A) Experimental timeline for mBH3@NPs administration in B16-F10-tumor-bearing mice. Mice were administered with mBH3@NPs three times, with a dosing interval of every 2 days (B) Average tumor volume curves for mice treated in the melanoma model ( n = 8). (C) Individual tumor volume curves for mice treated in the melanoma model ( n = 8). (D) Western blot analysis of Bax, Bak, Bcl-2, Bcl-x L , Mcl-1, Puma, Bim, caspase-3, and caspase-9 expression following different treatments. β-Actin was used as a loading control ( n = 3). (E) Representative images of tumor immunostaining and quantification for TUNEL (green)/DAPI (blue), CD8 (red)/DAPI (blue), and CRT (green)/DAPI (blue) at day 19 following the indicated treatments in the melanoma model. Scale bars, 200 μm. (F) HMGB1 expression in tumor tissues analyzed by ELISA following different treatments ( n = 5). (G) Heatmap of cytokine expression levels (GrB, IFN-γ, TNF-α, IL-12, and IL-6) in tumor tissues following different treatments ( n = 5). (H) Flow cytometric analysis of immune cell populations (T cells, CD8 + T cells, CD4 + T cells, Treg cells, NK cells, macrophages, M1-like TAMs, M2-like TAMs, and MDSCs) within tumors and CD11c + CD80 + CD86 + mature dendritic cells in lymph nodes ( n = 5). One-way ANOVA with Tukey’s multiple comparisons test was used for all statistical analyses. Data are presented as the mean ± SD. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; NS, not significant. See also and .

    Article Snippet: Human CD8 + T cells were isolated from PBMCs using the Human CD8 + T cell Negative Selection Kit (MedChemExpress, #HY-KO351).

    Techniques: Western Blot, Expressing, Control, Immunostaining, TUNEL Assay, Enzyme-linked Immunosorbent Assay

    mBH3@NPs promoted the recruitment of adoptively transferred T cells (A) Schematic of transwell migration assay using B16-OVA cells and preactivated OT-1 T cells. (B) Transwell migration assay of OT-I T cells (CFDA-labeled). Representative flow cytometry graphs and quantification of T cell infiltration relative fold changes (calculated as the ratio of bottom well-located OT-I T cells to counting beads) ( n = 3). (C) Experimental timeline for mBH3@NPs administration combined with adoptively transferred T cells in B16-OVA tumor-bearing mice. (D) Serial IVIS images and quantification of infiltrating Cy5-T cell signals in B16-OVA tumor-bearing mice ( n = 5). (E) Quantification of tumor-infiltrating endogenous T cells and transferred CD8 + T cells per million cells ( n = 5). (F) Heatmap of RNA-seq data for key interleukin family member, chemokine family member, and granzyme family member from harvested tumor tissues, plotted as Z score of normalized gene expression for each gene ( n = 3). (G) Heatmap of the differences in pathway activities scored by GSVA from harvested tumor tissues, plotted as Z score of normalized gene expression for each gene ( n = 3). (H) Validation of key cytokines and chemokines expression by multiplex cytokine assay. Tumor tissue was homogenized, and the cell supernatant of each treatment group was collected ( n = 4). One-way ANOVA with Tukey’s multiple comparisons test was used for all statistical analyses. Data are presented as the mean ± SD. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; NS, not significant. See also .

    Journal: Cell Reports Medicine

    Article Title: mRNA lipid-nanoparticle-mediated mitochondrial apoptosis augments adoptive T cell immunotherapy

    doi: 10.1016/j.xcrm.2026.102706

    Figure Lengend Snippet: mBH3@NPs promoted the recruitment of adoptively transferred T cells (A) Schematic of transwell migration assay using B16-OVA cells and preactivated OT-1 T cells. (B) Transwell migration assay of OT-I T cells (CFDA-labeled). Representative flow cytometry graphs and quantification of T cell infiltration relative fold changes (calculated as the ratio of bottom well-located OT-I T cells to counting beads) ( n = 3). (C) Experimental timeline for mBH3@NPs administration combined with adoptively transferred T cells in B16-OVA tumor-bearing mice. (D) Serial IVIS images and quantification of infiltrating Cy5-T cell signals in B16-OVA tumor-bearing mice ( n = 5). (E) Quantification of tumor-infiltrating endogenous T cells and transferred CD8 + T cells per million cells ( n = 5). (F) Heatmap of RNA-seq data for key interleukin family member, chemokine family member, and granzyme family member from harvested tumor tissues, plotted as Z score of normalized gene expression for each gene ( n = 3). (G) Heatmap of the differences in pathway activities scored by GSVA from harvested tumor tissues, plotted as Z score of normalized gene expression for each gene ( n = 3). (H) Validation of key cytokines and chemokines expression by multiplex cytokine assay. Tumor tissue was homogenized, and the cell supernatant of each treatment group was collected ( n = 4). One-way ANOVA with Tukey’s multiple comparisons test was used for all statistical analyses. Data are presented as the mean ± SD. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; NS, not significant. See also .

    Article Snippet: Human CD8 + T cells were isolated from PBMCs using the Human CD8 + T cell Negative Selection Kit (MedChemExpress, #HY-KO351).

    Techniques: Transwell Migration Assay, Labeling, Flow Cytometry, RNA Sequencing, Gene Expression, Biomarker Discovery, Expressing, Multiplex Assay, Cytokine Assay

    mBH3@NPs combined with adoptive T cell therapy improved the therapeutic effect (A) Experimental timeline for mBH3@NPs administration combined with adoptively transferred T cells from PMEL mice in B16-F10-tumor-bearing mice. (B) The average tumor volume curves for mice treated in the B16-F10-tumor-bearing mice model ( n = 7). (C) Survival analysis for mice treated in B16-F10-tumor-bearing mice model ( n = 7). Mice were humanely euthanized when tumor volume reached the predefined ethical endpoint (15 mm × 15 mm). The x axis indicates days post-mBH3@NPs administration. (D) Experimental timeline for mBH3@NPs administration combined with adoptive transfered T cells from OT-1 mice in B16-OVA-tumor-bearing mice. (E) The average tumor volume curves for mice treated in B16-OVA-tumor-bearing mice model ( n = 7). (F) Survival analysis for mice treated in B16-OVA-tumor-bearing mice model ( n = 7). Mice were humanely euthanized when tumor volume reached the predefined ethical endpoint (15 mm × 15 mm). (G) Representative flow cytometry plot and quantification of CD44 and CD62L expression levels gated on total CD8 + T cells (upper panel) and the percentage of endogenous (CD90.2) CD8 + T cells and transferred PMEL (CD90.1) CD8 + T cells gated on total CD8 + T cells (lower panel) ( n = 5). (H) Representative flow cytometry plot and quantification of the expression level of granzyme B (GrB) and perforin in total CD8 + T cells from tumors and spleens ( n = 5). (I) Representative flow cytometry plot and quantification of the expression level of PD-1 and LAG-3 in both endogenous and transferred PMEL CD8 + T cells from tumors ( n = 5). (J) Immunofluorescence evaluation of CD8 + T cell exhaustion, effector, and memory function. Upper panel: representative immunofluorescence images of tumors from different groups for analyzing the exhaustion (PD-1 + , cyan) and effector function (IFN-γ + , green) of CD8 + T cells (CD8 + , pink). Scale bars, 100 μm. Lower panel: representative immunofluorescence images of tumor-infiltrating CD8 + T cells (CD8 + , green) along with markers for activation (CD69 + , red) and memory differentiation (CD44 + , pink; CCR7 + , cyan). Scale bars, 200 μm ( n = 3). One-way ANOVA with Tukey’s multiple comparisons test was used for all statistical analyses. Data are presented as the mean ± SD. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; NS, not significant. See also .

    Journal: Cell Reports Medicine

    Article Title: mRNA lipid-nanoparticle-mediated mitochondrial apoptosis augments adoptive T cell immunotherapy

    doi: 10.1016/j.xcrm.2026.102706

    Figure Lengend Snippet: mBH3@NPs combined with adoptive T cell therapy improved the therapeutic effect (A) Experimental timeline for mBH3@NPs administration combined with adoptively transferred T cells from PMEL mice in B16-F10-tumor-bearing mice. (B) The average tumor volume curves for mice treated in the B16-F10-tumor-bearing mice model ( n = 7). (C) Survival analysis for mice treated in B16-F10-tumor-bearing mice model ( n = 7). Mice were humanely euthanized when tumor volume reached the predefined ethical endpoint (15 mm × 15 mm). The x axis indicates days post-mBH3@NPs administration. (D) Experimental timeline for mBH3@NPs administration combined with adoptive transfered T cells from OT-1 mice in B16-OVA-tumor-bearing mice. (E) The average tumor volume curves for mice treated in B16-OVA-tumor-bearing mice model ( n = 7). (F) Survival analysis for mice treated in B16-OVA-tumor-bearing mice model ( n = 7). Mice were humanely euthanized when tumor volume reached the predefined ethical endpoint (15 mm × 15 mm). (G) Representative flow cytometry plot and quantification of CD44 and CD62L expression levels gated on total CD8 + T cells (upper panel) and the percentage of endogenous (CD90.2) CD8 + T cells and transferred PMEL (CD90.1) CD8 + T cells gated on total CD8 + T cells (lower panel) ( n = 5). (H) Representative flow cytometry plot and quantification of the expression level of granzyme B (GrB) and perforin in total CD8 + T cells from tumors and spleens ( n = 5). (I) Representative flow cytometry plot and quantification of the expression level of PD-1 and LAG-3 in both endogenous and transferred PMEL CD8 + T cells from tumors ( n = 5). (J) Immunofluorescence evaluation of CD8 + T cell exhaustion, effector, and memory function. Upper panel: representative immunofluorescence images of tumors from different groups for analyzing the exhaustion (PD-1 + , cyan) and effector function (IFN-γ + , green) of CD8 + T cells (CD8 + , pink). Scale bars, 100 μm. Lower panel: representative immunofluorescence images of tumor-infiltrating CD8 + T cells (CD8 + , green) along with markers for activation (CD69 + , red) and memory differentiation (CD44 + , pink; CCR7 + , cyan). Scale bars, 200 μm ( n = 3). One-way ANOVA with Tukey’s multiple comparisons test was used for all statistical analyses. Data are presented as the mean ± SD. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; NS, not significant. See also .

    Article Snippet: Human CD8 + T cells were isolated from PBMCs using the Human CD8 + T cell Negative Selection Kit (MedChemExpress, #HY-KO351).

    Techniques: Flow Cytometry, Expressing, Immunofluorescence, Activation Assay

    Combination therapy drove CD8 + T cells toward memory-like states with expanded TCR diversity (A) Experimental design for scRNA-seq. Mice were treated with ACT alone or combined with mBim@NPs and mPuma@NPs as shown ( n = 3); CD3 + T cell isolated from mice for scRNA-seq. (B) UMAP representation and distribution of 68,176 cells of the CD8 + T cell atlas in each experimental condition, colored by 12 CD8 + T cell subtypes annotated in this study. (C) UMAP representation of the CD8 + T cells atlas colored by each experimental condition. (D) A dot plot showing the expression levels of marker genes in each CD8 + T cell subtype. The color of the dots indicates the average scaled expression level, and the size of the dots indicates the percentage of cells expressing the gene in each subtype. (E) Indicated expression of gene marker associated with memory ( Tcf7 , Il7r , Sell , and Ccl5 ), exhaustion ( Pdcd1 , Tigit , and Lag3 ), and cytotoxicity ( Gzmb , Ifng , and Prf1 ) in the UMAP plot. (F) A dot plot showing the expression levels of marker genes in E. The color of the dots indicates the average scaled expression level, and the size of the dots indicates the percentage of cells expressing the gene in each subtype. (G) Volcano plots of log 2 (fold change) and log 10 (adjusted FDR value) of differentially expressed genes in proliferating T and Gzmk + T rm from mice treated with ACT monotherapy and combination therapy with mBim@NPs. (H) Shannon entropy of the T ex , proliferating T, T naive , and Gzmk + T rm subsets across different treatment groups (T: ACT monotherapy; TP: ACT + mPuma@NPs; TB: ACT + mBim@NPs). (I) Shannon entropy of total CD8 + T cells and CD4 + T cells across different treatment groups. (J) A heatmap of normalized Shannon entropy for CD8 + T cell subsets across different treatment groups. See also .

    Journal: Cell Reports Medicine

    Article Title: mRNA lipid-nanoparticle-mediated mitochondrial apoptosis augments adoptive T cell immunotherapy

    doi: 10.1016/j.xcrm.2026.102706

    Figure Lengend Snippet: Combination therapy drove CD8 + T cells toward memory-like states with expanded TCR diversity (A) Experimental design for scRNA-seq. Mice were treated with ACT alone or combined with mBim@NPs and mPuma@NPs as shown ( n = 3); CD3 + T cell isolated from mice for scRNA-seq. (B) UMAP representation and distribution of 68,176 cells of the CD8 + T cell atlas in each experimental condition, colored by 12 CD8 + T cell subtypes annotated in this study. (C) UMAP representation of the CD8 + T cells atlas colored by each experimental condition. (D) A dot plot showing the expression levels of marker genes in each CD8 + T cell subtype. The color of the dots indicates the average scaled expression level, and the size of the dots indicates the percentage of cells expressing the gene in each subtype. (E) Indicated expression of gene marker associated with memory ( Tcf7 , Il7r , Sell , and Ccl5 ), exhaustion ( Pdcd1 , Tigit , and Lag3 ), and cytotoxicity ( Gzmb , Ifng , and Prf1 ) in the UMAP plot. (F) A dot plot showing the expression levels of marker genes in E. The color of the dots indicates the average scaled expression level, and the size of the dots indicates the percentage of cells expressing the gene in each subtype. (G) Volcano plots of log 2 (fold change) and log 10 (adjusted FDR value) of differentially expressed genes in proliferating T and Gzmk + T rm from mice treated with ACT monotherapy and combination therapy with mBim@NPs. (H) Shannon entropy of the T ex , proliferating T, T naive , and Gzmk + T rm subsets across different treatment groups (T: ACT monotherapy; TP: ACT + mPuma@NPs; TB: ACT + mBim@NPs). (I) Shannon entropy of total CD8 + T cells and CD4 + T cells across different treatment groups. (J) A heatmap of normalized Shannon entropy for CD8 + T cell subsets across different treatment groups. See also .

    Article Snippet: Human CD8 + T cells were isolated from PBMCs using the Human CD8 + T cell Negative Selection Kit (MedChemExpress, #HY-KO351).

    Techniques: Isolation, Expressing, Marker