biotin cd3e (clone 145-2c11) ebioscience antibody Search Results


cd3  (Miltenyi Biotec)
99
Miltenyi Biotec cd3
Intratumoral treatment with agonistic 41BB antibodies increases CD8 T cell infiltration. A and B, Mass of MC38 and Panc02-ZsGOVA tumors harvested 7 days after initial α41BB treatment. C and D, Frequency of CD4 and CD8 TILs in MC38 and Panc02-ZsGOVA tumors 7 days after initial α41BB treatment. E, Mice with MC38 tumors were treated with isotype or α41BB antibodies in combination with CD8 depleting antibodies. Tumor growth is shown (n=5 mice per group). F, TILs were isolated from MC38 tumors treated with intratumoral isotype or α41BB antibodies. TILs were cultured with immobilized <t>anti-CD3</t> antibody or co-cultured with irradiated MC38 or B16 tumor lines for 48hrs. IFN-γ was measured from supernatants. One of two representative experiments are shown. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001.
Cd3, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 99 stars, based on 1 article reviews
cd3 - by Bioz Stars, 2026-03
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pe/ cy7 conjugated anti cd3e  (Becton Dickinson)
90
Becton Dickinson pe/ cy7 conjugated anti cd3e
Intratumoral treatment with agonistic 41BB antibodies increases CD8 T cell infiltration. A and B, Mass of MC38 and Panc02-ZsGOVA tumors harvested 7 days after initial α41BB treatment. C and D, Frequency of CD4 and CD8 TILs in MC38 and Panc02-ZsGOVA tumors 7 days after initial α41BB treatment. E, Mice with MC38 tumors were treated with isotype or α41BB antibodies in combination with CD8 depleting antibodies. Tumor growth is shown (n=5 mice per group). F, TILs were isolated from MC38 tumors treated with intratumoral isotype or α41BB antibodies. TILs were cultured with immobilized <t>anti-CD3</t> antibody or co-cultured with irradiated MC38 or B16 tumor lines for 48hrs. IFN-γ was measured from supernatants. One of two representative experiments are shown. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001.
Pe/ Cy7 Conjugated Anti Cd3e, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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cd3e monoclonal antibody pe-cyanine5.5  (Thermo Fisher)
90
Thermo Fisher cd3e monoclonal antibody pe-cyanine5.5
Intratumoral treatment with agonistic 41BB antibodies increases CD8 T cell infiltration. A and B, Mass of MC38 and Panc02-ZsGOVA tumors harvested 7 days after initial α41BB treatment. C and D, Frequency of CD4 and CD8 TILs in MC38 and Panc02-ZsGOVA tumors 7 days after initial α41BB treatment. E, Mice with MC38 tumors were treated with isotype or α41BB antibodies in combination with CD8 depleting antibodies. Tumor growth is shown (n=5 mice per group). F, TILs were isolated from MC38 tumors treated with intratumoral isotype or α41BB antibodies. TILs were cultured with immobilized <t>anti-CD3</t> antibody or co-cultured with irradiated MC38 or B16 tumor lines for 48hrs. IFN-γ was measured from supernatants. One of two representative experiments are shown. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001.
Cd3e Monoclonal Antibody Pe Cyanine5.5, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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cd3e  (Cytek Biosciences)
93
Cytek Biosciences cd3e
At 8dpi, pooled splenocytes (by genotype) from age matched, sex matched mice were sorted for live CD45+ cells and submitted for single cell RNA sequencing (scRNAseq) on the 10x genomic platform (A). Relative gene expression of Ptprc indicating immune cell in aggregated data set (both genotypes) (B). Relative gene expression to located immune cell clusters of myeloid Csfr1 , neutrophil Ly6g , B cell Cd19, and T cell <t>Cd3e</t> (C). Using Loupe software, each population was reclustered to group like cell subsets among the larger cell type and the proportions compared between genotypes. The frequency of each cluster among total cell population is listed on the graph. Myeloid cell cluster, based on Csf1r expression (D), Neutrophil clusters, based on Ly6g expression (E), and B cell cluster, based on Cd19 expression (F). T cells were further broken down into CD4 T cells (expression of Cd3e >0, Mt2 <10, Cd4 >0) (G) and CD8 T cells (expression of Cd3e >0, Mt2 <10, Cd8b1 >0) (H). tSNE plots highlighting Cd4 (G) and Cd8b1 (H) as well as breakdown of cluster for each T cell type in -WT and - 619WW cells. Corresponding quantification of proportion of either CD4 T cells or CD8 T cell subsets is next to tSNE plot.
Cd3e, supplied by Cytek Biosciences, 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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pe conjugated anti-cd3e  (Thermo Fisher)
90
Thermo Fisher pe conjugated anti-cd3e
At 8dpi, pooled splenocytes (by genotype) from age matched, sex matched mice were sorted for live CD45+ cells and submitted for single cell RNA sequencing (scRNAseq) on the 10x genomic platform (A). Relative gene expression of Ptprc indicating immune cell in aggregated data set (both genotypes) (B). Relative gene expression to located immune cell clusters of myeloid Csfr1 , neutrophil Ly6g , B cell Cd19, and T cell <t>Cd3e</t> (C). Using Loupe software, each population was reclustered to group like cell subsets among the larger cell type and the proportions compared between genotypes. The frequency of each cluster among total cell population is listed on the graph. Myeloid cell cluster, based on Csf1r expression (D), Neutrophil clusters, based on Ly6g expression (E), and B cell cluster, based on Cd19 expression (F). T cells were further broken down into CD4 T cells (expression of Cd3e >0, Mt2 <10, Cd4 >0) (G) and CD8 T cells (expression of Cd3e >0, Mt2 <10, Cd8b1 >0) (H). tSNE plots highlighting Cd4 (G) and Cd8b1 (H) as well as breakdown of cluster for each T cell type in -WT and - 619WW cells. Corresponding quantification of proportion of either CD4 T cells or CD8 T cell subsets is next to tSNE plot.
Pe Conjugated Anti Cd3e, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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pe conjugated antibodies against lineage + markers mac1 m1/70  (Thermo Fisher)
90
Thermo Fisher pe conjugated antibodies against lineage + markers mac1 m1/70
At 8dpi, pooled splenocytes (by genotype) from age matched, sex matched mice were sorted for live CD45+ cells and submitted for single cell RNA sequencing (scRNAseq) on the 10x genomic platform (A). Relative gene expression of Ptprc indicating immune cell in aggregated data set (both genotypes) (B). Relative gene expression to located immune cell clusters of myeloid Csfr1 , neutrophil Ly6g , B cell Cd19, and T cell <t>Cd3e</t> (C). Using Loupe software, each population was reclustered to group like cell subsets among the larger cell type and the proportions compared between genotypes. The frequency of each cluster among total cell population is listed on the graph. Myeloid cell cluster, based on Csf1r expression (D), Neutrophil clusters, based on Ly6g expression (E), and B cell cluster, based on Cd19 expression (F). T cells were further broken down into CD4 T cells (expression of Cd3e >0, Mt2 <10, Cd4 >0) (G) and CD8 T cells (expression of Cd3e >0, Mt2 <10, Cd8b1 >0) (H). tSNE plots highlighting Cd4 (G) and Cd8b1 (H) as well as breakdown of cluster for each T cell type in -WT and - 619WW cells. Corresponding quantification of proportion of either CD4 T cells or CD8 T cell subsets is next to tSNE plot.
Pe Conjugated Antibodies Against Lineage + Markers Mac1 M1/70, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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anti mouse cd3e 145 2c11 ebioscience  (Thermo Fisher)
86
Thermo Fisher anti mouse cd3e 145 2c11 ebioscience
At 8dpi, pooled splenocytes (by genotype) from age matched, sex matched mice were sorted for live CD45+ cells and submitted for single cell RNA sequencing (scRNAseq) on the 10x genomic platform (A). Relative gene expression of Ptprc indicating immune cell in aggregated data set (both genotypes) (B). Relative gene expression to located immune cell clusters of myeloid Csfr1 , neutrophil Ly6g , B cell Cd19, and T cell <t>Cd3e</t> (C). Using Loupe software, each population was reclustered to group like cell subsets among the larger cell type and the proportions compared between genotypes. The frequency of each cluster among total cell population is listed on the graph. Myeloid cell cluster, based on Csf1r expression (D), Neutrophil clusters, based on Ly6g expression (E), and B cell cluster, based on Cd19 expression (F). T cells were further broken down into CD4 T cells (expression of Cd3e >0, Mt2 <10, Cd4 >0) (G) and CD8 T cells (expression of Cd3e >0, Mt2 <10, Cd8b1 >0) (H). tSNE plots highlighting Cd4 (G) and Cd8b1 (H) as well as breakdown of cluster for each T cell type in -WT and - 619WW cells. Corresponding quantification of proportion of either CD4 T cells or CD8 T cell subsets is next to tSNE plot.
Anti Mouse Cd3e 145 2c11 Ebioscience, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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fluorescein isothiocyanate (fitc)-conjugated hamster anti-mouse cd3e (145-2c11)  (Thermo Fisher)
90
Thermo Fisher fluorescein isothiocyanate (fitc)-conjugated hamster anti-mouse cd3e (145-2c11)
At 8dpi, pooled splenocytes (by genotype) from age matched, sex matched mice were sorted for live CD45+ cells and submitted for single cell RNA sequencing (scRNAseq) on the 10x genomic platform (A). Relative gene expression of Ptprc indicating immune cell in aggregated data set (both genotypes) (B). Relative gene expression to located immune cell clusters of myeloid Csfr1 , neutrophil Ly6g , B cell Cd19, and T cell <t>Cd3e</t> (C). Using Loupe software, each population was reclustered to group like cell subsets among the larger cell type and the proportions compared between genotypes. The frequency of each cluster among total cell population is listed on the graph. Myeloid cell cluster, based on Csf1r expression (D), Neutrophil clusters, based on Ly6g expression (E), and B cell cluster, based on Cd19 expression (F). T cells were further broken down into CD4 T cells (expression of Cd3e >0, Mt2 <10, Cd4 >0) (G) and CD8 T cells (expression of Cd3e >0, Mt2 <10, Cd8b1 >0) (H). tSNE plots highlighting Cd4 (G) and Cd8b1 (H) as well as breakdown of cluster for each T cell type in -WT and - 619WW cells. Corresponding quantification of proportion of either CD4 T cells or CD8 T cell subsets is next to tSNE plot.
Fluorescein Isothiocyanate (Fitc) Conjugated Hamster Anti Mouse Cd3e (145 2c11), supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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anti cd3e clone 145 2c11 standard biotools  (fluidigm)
95
fluidigm anti cd3e clone 145 2c11 standard biotools
At 8dpi, pooled splenocytes (by genotype) from age matched, sex matched mice were sorted for live CD45+ cells and submitted for single cell RNA sequencing (scRNAseq) on the 10x genomic platform (A). Relative gene expression of Ptprc indicating immune cell in aggregated data set (both genotypes) (B). Relative gene expression to located immune cell clusters of myeloid Csfr1 , neutrophil Ly6g , B cell Cd19, and T cell <t>Cd3e</t> (C). Using Loupe software, each population was reclustered to group like cell subsets among the larger cell type and the proportions compared between genotypes. The frequency of each cluster among total cell population is listed on the graph. Myeloid cell cluster, based on Csf1r expression (D), Neutrophil clusters, based on Ly6g expression (E), and B cell cluster, based on Cd19 expression (F). T cells were further broken down into CD4 T cells (expression of Cd3e >0, Mt2 <10, Cd4 >0) (G) and CD8 T cells (expression of Cd3e >0, Mt2 <10, Cd8b1 >0) (H). tSNE plots highlighting Cd4 (G) and Cd8b1 (H) as well as breakdown of cluster for each T cell type in -WT and - 619WW cells. Corresponding quantification of proportion of either CD4 T cells or CD8 T cell subsets is next to tSNE plot.
Anti Cd3e Clone 145 2c11 Standard Biotools, supplied by fluidigm, 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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cd3e- pe cy5  (Thermo Fisher)
90
Thermo Fisher cd3e- pe cy5
At 8dpi, pooled splenocytes (by genotype) from age matched, sex matched mice were sorted for live CD45+ cells and submitted for single cell RNA sequencing (scRNAseq) on the 10x genomic platform (A). Relative gene expression of Ptprc indicating immune cell in aggregated data set (both genotypes) (B). Relative gene expression to located immune cell clusters of myeloid Csfr1 , neutrophil Ly6g , B cell Cd19, and T cell <t>Cd3e</t> (C). Using Loupe software, each population was reclustered to group like cell subsets among the larger cell type and the proportions compared between genotypes. The frequency of each cluster among total cell population is listed on the graph. Myeloid cell cluster, based on Csf1r expression (D), Neutrophil clusters, based on Ly6g expression (E), and B cell cluster, based on Cd19 expression (F). T cells were further broken down into CD4 T cells (expression of Cd3e >0, Mt2 <10, Cd4 >0) (G) and CD8 T cells (expression of Cd3e >0, Mt2 <10, Cd8b1 >0) (H). tSNE plots highlighting Cd4 (G) and Cd8b1 (H) as well as breakdown of cluster for each T cell type in -WT and - 619WW cells. Corresponding quantification of proportion of either CD4 T cells or CD8 T cell subsets is next to tSNE plot.
Cd3e Pe Cy5, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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anti-cd3e 145-2c11  (Immunostep)
90
Immunostep anti-cd3e 145-2c11
At 8dpi, pooled splenocytes (by genotype) from age matched, sex matched mice were sorted for live CD45+ cells and submitted for single cell RNA sequencing (scRNAseq) on the 10x genomic platform (A). Relative gene expression of Ptprc indicating immune cell in aggregated data set (both genotypes) (B). Relative gene expression to located immune cell clusters of myeloid Csfr1 , neutrophil Ly6g , B cell Cd19, and T cell <t>Cd3e</t> (C). Using Loupe software, each population was reclustered to group like cell subsets among the larger cell type and the proportions compared between genotypes. The frequency of each cluster among total cell population is listed on the graph. Myeloid cell cluster, based on Csf1r expression (D), Neutrophil clusters, based on Ly6g expression (E), and B cell cluster, based on Cd19 expression (F). T cells were further broken down into CD4 T cells (expression of Cd3e >0, Mt2 <10, Cd4 >0) (G) and CD8 T cells (expression of Cd3e >0, Mt2 <10, Cd8b1 >0) (H). tSNE plots highlighting Cd4 (G) and Cd8b1 (H) as well as breakdown of cluster for each T cell type in -WT and - 619WW cells. Corresponding quantification of proportion of either CD4 T cells or CD8 T cell subsets is next to tSNE plot.
Anti Cd3e 145 2c11, supplied by Immunostep, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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hamster anti-mouse cd3e-pe-cy7  (Thermo Fisher)
90
Thermo Fisher hamster anti-mouse cd3e-pe-cy7
At 8dpi, pooled splenocytes (by genotype) from age matched, sex matched mice were sorted for live CD45+ cells and submitted for single cell RNA sequencing (scRNAseq) on the 10x genomic platform (A). Relative gene expression of Ptprc indicating immune cell in aggregated data set (both genotypes) (B). Relative gene expression to located immune cell clusters of myeloid Csfr1 , neutrophil Ly6g , B cell Cd19, and T cell <t>Cd3e</t> (C). Using Loupe software, each population was reclustered to group like cell subsets among the larger cell type and the proportions compared between genotypes. The frequency of each cluster among total cell population is listed on the graph. Myeloid cell cluster, based on Csf1r expression (D), Neutrophil clusters, based on Ly6g expression (E), and B cell cluster, based on Cd19 expression (F). T cells were further broken down into CD4 T cells (expression of Cd3e >0, Mt2 <10, Cd4 >0) (G) and CD8 T cells (expression of Cd3e >0, Mt2 <10, Cd8b1 >0) (H). tSNE plots highlighting Cd4 (G) and Cd8b1 (H) as well as breakdown of cluster for each T cell type in -WT and - 619WW cells. Corresponding quantification of proportion of either CD4 T cells or CD8 T cell subsets is next to tSNE plot.
Hamster Anti Mouse Cd3e Pe Cy7, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Image Search Results


Intratumoral treatment with agonistic 41BB antibodies increases CD8 T cell infiltration. A and B, Mass of MC38 and Panc02-ZsGOVA tumors harvested 7 days after initial α41BB treatment. C and D, Frequency of CD4 and CD8 TILs in MC38 and Panc02-ZsGOVA tumors 7 days after initial α41BB treatment. E, Mice with MC38 tumors were treated with isotype or α41BB antibodies in combination with CD8 depleting antibodies. Tumor growth is shown (n=5 mice per group). F, TILs were isolated from MC38 tumors treated with intratumoral isotype or α41BB antibodies. TILs were cultured with immobilized anti-CD3 antibody or co-cultured with irradiated MC38 or B16 tumor lines for 48hrs. IFN-γ was measured from supernatants. One of two representative experiments are shown. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001.

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Intratumoral activation of 41BB co-stimulatory signals enhances CD8 T cell expansion and modulates tumor-infiltrating myeloid cells

doi: 10.4049/jimmunol.2000759

Figure Lengend Snippet: Intratumoral treatment with agonistic 41BB antibodies increases CD8 T cell infiltration. A and B, Mass of MC38 and Panc02-ZsGOVA tumors harvested 7 days after initial α41BB treatment. C and D, Frequency of CD4 and CD8 TILs in MC38 and Panc02-ZsGOVA tumors 7 days after initial α41BB treatment. E, Mice with MC38 tumors were treated with isotype or α41BB antibodies in combination with CD8 depleting antibodies. Tumor growth is shown (n=5 mice per group). F, TILs were isolated from MC38 tumors treated with intratumoral isotype or α41BB antibodies. TILs were cultured with immobilized anti-CD3 antibody or co-cultured with irradiated MC38 or B16 tumor lines for 48hrs. IFN-γ was measured from supernatants. One of two representative experiments are shown. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001.

Article Snippet: For surface staining of murine specimens, cells were stained in FACS buffer with the following antibodies: CD3 (145–2C11), CD4 (GK1.5), CD8 (53–6.7), CD11b (M1/70), Ly6G (1A8), Ly6C (HK1.4), F4/80 (BM8), CD11c (N418), MHCII (M5/114.15.2), CD80 (16–10A1), CD86 (GL-1), PD-1 (29F.1A12) (all from BioLegend); 41BB (17B5–1H1, Miltenyi Biotec).

Techniques: Isolation, Cell Culture, Irradiation

41BBL expression on APCs alters the capacity to prime TILs. TILs from a melanoma patient were cultured with αCD3, autologous tumor cells at a 1:1 ratio, or tumor-lysate pulsed 41BBL APCs at a 1:10 ratio for 72hrs. A and B, Heatmap representing cytokine abundance in supernatants from cell cultures were collected at 72hrs. Numerical values are indicated for each parameter with its respective condition. Cytokine production by TIL stimulated with αCD3 +/− urelumab (A). Cytokines in TIL-tumor co-cultures +/− α41BB or MHC-I blocking antibodies (B). C, 41BBL APCs were generated and then pulsed with autologous tumor lysate in the presence of GMCSF. Pulsed APCs were then seeded in culture wells with or without α41BB. D, Cytokines were measured in the supernatants of TIL-APC co-cultures incubated with α41BB and/or α 41BBL. TIL only condition is the same data from (B); APCs only from (C). Statistics are indicated for cytokines that are higher than both TILs alone and APCs alone conditions. E, Fold change in cytokine induction vs. TIL+APCs in co-cultures from (D). Dotted line represents the basal induction of cytokines in TIL-APC co-cultures. F, TIL proliferation in co-cultures was determined in the final 18hrs of the culture by 3H thymidine incorporation in the presence of soluble α41BB and/or soluble α41BBL. Dotted line represents basal TIL proliferation without additional stimulation. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001. Significance was determined by two-tailed t-test or 2-way ANOVA with Dunnett’s multiple comparisons. ND = not detected.

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Intratumoral activation of 41BB co-stimulatory signals enhances CD8 T cell expansion and modulates tumor-infiltrating myeloid cells

doi: 10.4049/jimmunol.2000759

Figure Lengend Snippet: 41BBL expression on APCs alters the capacity to prime TILs. TILs from a melanoma patient were cultured with αCD3, autologous tumor cells at a 1:1 ratio, or tumor-lysate pulsed 41BBL APCs at a 1:10 ratio for 72hrs. A and B, Heatmap representing cytokine abundance in supernatants from cell cultures were collected at 72hrs. Numerical values are indicated for each parameter with its respective condition. Cytokine production by TIL stimulated with αCD3 +/− urelumab (A). Cytokines in TIL-tumor co-cultures +/− α41BB or MHC-I blocking antibodies (B). C, 41BBL APCs were generated and then pulsed with autologous tumor lysate in the presence of GMCSF. Pulsed APCs were then seeded in culture wells with or without α41BB. D, Cytokines were measured in the supernatants of TIL-APC co-cultures incubated with α41BB and/or α 41BBL. TIL only condition is the same data from (B); APCs only from (C). Statistics are indicated for cytokines that are higher than both TILs alone and APCs alone conditions. E, Fold change in cytokine induction vs. TIL+APCs in co-cultures from (D). Dotted line represents the basal induction of cytokines in TIL-APC co-cultures. F, TIL proliferation in co-cultures was determined in the final 18hrs of the culture by 3H thymidine incorporation in the presence of soluble α41BB and/or soluble α41BBL. Dotted line represents basal TIL proliferation without additional stimulation. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001. Significance was determined by two-tailed t-test or 2-way ANOVA with Dunnett’s multiple comparisons. ND = not detected.

Article Snippet: For surface staining of murine specimens, cells were stained in FACS buffer with the following antibodies: CD3 (145–2C11), CD4 (GK1.5), CD8 (53–6.7), CD11b (M1/70), Ly6G (1A8), Ly6C (HK1.4), F4/80 (BM8), CD11c (N418), MHCII (M5/114.15.2), CD80 (16–10A1), CD86 (GL-1), PD-1 (29F.1A12) (all from BioLegend); 41BB (17B5–1H1, Miltenyi Biotec).

Techniques: Expressing, Cell Culture, Blocking Assay, Generated, Incubation, Two Tailed Test

At 8dpi, pooled splenocytes (by genotype) from age matched, sex matched mice were sorted for live CD45+ cells and submitted for single cell RNA sequencing (scRNAseq) on the 10x genomic platform (A). Relative gene expression of Ptprc indicating immune cell in aggregated data set (both genotypes) (B). Relative gene expression to located immune cell clusters of myeloid Csfr1 , neutrophil Ly6g , B cell Cd19, and T cell Cd3e (C). Using Loupe software, each population was reclustered to group like cell subsets among the larger cell type and the proportions compared between genotypes. The frequency of each cluster among total cell population is listed on the graph. Myeloid cell cluster, based on Csf1r expression (D), Neutrophil clusters, based on Ly6g expression (E), and B cell cluster, based on Cd19 expression (F). T cells were further broken down into CD4 T cells (expression of Cd3e >0, Mt2 <10, Cd4 >0) (G) and CD8 T cells (expression of Cd3e >0, Mt2 <10, Cd8b1 >0) (H). tSNE plots highlighting Cd4 (G) and Cd8b1 (H) as well as breakdown of cluster for each T cell type in -WT and - 619WW cells. Corresponding quantification of proportion of either CD4 T cells or CD8 T cell subsets is next to tSNE plot.

Journal: bioRxiv

Article Title: Autoimmunity associated allele of tyrosine phosphatase gene PTPN22 enhances anti-viral immunity

doi: 10.1101/2023.06.28.546768

Figure Lengend Snippet: At 8dpi, pooled splenocytes (by genotype) from age matched, sex matched mice were sorted for live CD45+ cells and submitted for single cell RNA sequencing (scRNAseq) on the 10x genomic platform (A). Relative gene expression of Ptprc indicating immune cell in aggregated data set (both genotypes) (B). Relative gene expression to located immune cell clusters of myeloid Csfr1 , neutrophil Ly6g , B cell Cd19, and T cell Cd3e (C). Using Loupe software, each population was reclustered to group like cell subsets among the larger cell type and the proportions compared between genotypes. The frequency of each cluster among total cell population is listed on the graph. Myeloid cell cluster, based on Csf1r expression (D), Neutrophil clusters, based on Ly6g expression (E), and B cell cluster, based on Cd19 expression (F). T cells were further broken down into CD4 T cells (expression of Cd3e >0, Mt2 <10, Cd4 >0) (G) and CD8 T cells (expression of Cd3e >0, Mt2 <10, Cd8b1 >0) (H). tSNE plots highlighting Cd4 (G) and Cd8b1 (H) as well as breakdown of cluster for each T cell type in -WT and - 619WW cells. Corresponding quantification of proportion of either CD4 T cells or CD8 T cell subsets is next to tSNE plot.

Article Snippet: Antibodies used in various combinations (depending on experiment) are as follows: Ghost Viability Dye (v510, Tonbo Biosciences, 1:1000 dilution), CD3e (PE-Cy5/AF532, Tonbo Bioscience/Thermo Fischer, 1:200, clone 145-2C11), CD4 (PerCP/BV605, Tonbo Bioscience/Biolegend, 1:200, clone RM4-5), CD8α (APC-Cy7/APC-H7/APC, BD Biosciences, 1:200, clone 53-6.7), CD11c (PE-Cy5.5, Thermo Fisher, 1:100, clone N418), CD11b (PerCP-Cy5.5, Biolegend, 1:200, clone M1/70), F4/80 (Pacific Orange, Thermo Fisher, 1:100, clone BM8), IFNγ (AF647, Biolegend, 1:100, clone XMG1.2), TNFα (PE-Cy7, Beiolegend, 1:100, clone MP6-XT22), IL-2 (PE/BV421, Biolegend, 1:50, clone JES6-5H4), PDCA-1 (Pacific Blue, Biolegend, 1:200, clone 129C1), CD80 (BV421, 1:200, clone 16-10A1), CD86 (BV605, Biolegend, 1:200, clone GL1), PD-1 (PE-Cy7, Tonbo Biosciences, 1:200, clone J43.1), PD- L1 (PE/BV711, Tonbo Biosciences/Biolegend, 1:100, clone 10F.9G2), CD44 (AF700, Biolegend, 1:200, clone IM7), CD62L (FITC, Tonbo Biosciences, 1:100, MEL-14), Ly6C (BV785, Biolegend, 1:200, clone HK1.4), Ly6G (PE-eFlour610, Invitrogen, 1:200, clone IA8), CD206 (AF647, Biolegend, 1:100, clone CO68C2), CD209b (APC, Tonbo Biosciences, 1:200, clone 22D1), NK1.1 (FITC, Biolegend, 1:100, clone PK136), CD19 (BV711, Biolegend, 1:400, clone 6D5), B220 (APC-Cy5.5, Invitrogen, 1:200, clone RA3-6B2), MHC II I-Ab (FITC, Biolegend, 1:200, clone AF6-120.1), CXCR5 (BV605, Biolegend, 1:100, clone L138D7), Tbet (APC/AF647, Biolegend, 1:200), Foxp3 (PE, Invitrogen, 1:100, clone FJK-16s), LCMV-NP (self-conjugated to AF488 using Thermo Fisher AF488 labeling kit per manufacturer’s instructions, BioXcell, 1:50, clone VL-4).

Techniques: RNA Sequencing, Gene Expression, Software, Expressing