anti cd3 apc efluor780  (Thermo Fisher)


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    Thermo Fisher anti cd3 apc efluor780
    CAR T cells use an antibody fragment (such as a single-chain variable fragment as shown here) as the binding domain—which is fused to CD3ζ and often contains additional co-stimulatory domains—that targets a cell surface protein. TRuC T cells also use an antibody fragment (such as a single-domain antibody as shown here) that targets a cell surface protein; however, this antibody fragment is fused to another part of the <t>TCR:CD3</t> complex—shown here, it is fused to the CD3ε, using natural T-cell signaling pathways. FvTCR T cells replace the variable domains of the TCR with the light and heavy chains of the antibody. HiT T cells use TCR binding domains to target cell surface proteins directly and therefore use natural cell signaling pathways. TCR T cells use a TCR binding domain to specifically target a peptide presented by the HLA molecule. CAR, chimeric antigen receptor; FvTCR, variable fragment T-cell receptor; HiT, human leukocyte antigen–independent T-cell receptor; HLA, human leukocyte antigen; TCR, T-cell receptor; TRuC, T-cell receptor fusion construct.
    Anti Cd3 Apc Efluor780, 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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    1) Product Images from "Selection, engineering, and in vivo testing of a human leukocyte antigen–independent T-cell receptor recognizing human mesothelin"

    Article Title: Selection, engineering, and in vivo testing of a human leukocyte antigen–independent T-cell receptor recognizing human mesothelin

    Journal: PLOS ONE

    doi: 10.1371/journal.pone.0301175

    CAR T cells use an antibody fragment (such as a single-chain variable fragment as shown here) as the binding domain—which is fused to CD3ζ and often contains additional co-stimulatory domains—that targets a cell surface protein. TRuC T cells also use an antibody fragment (such as a single-domain antibody as shown here) that targets a cell surface protein; however, this antibody fragment is fused to another part of the TCR:CD3 complex—shown here, it is fused to the CD3ε, using natural T-cell signaling pathways. FvTCR T cells replace the variable domains of the TCR with the light and heavy chains of the antibody. HiT T cells use TCR binding domains to target cell surface proteins directly and therefore use natural cell signaling pathways. TCR T cells use a TCR binding domain to specifically target a peptide presented by the HLA molecule. CAR, chimeric antigen receptor; FvTCR, variable fragment T-cell receptor; HiT, human leukocyte antigen–independent T-cell receptor; HLA, human leukocyte antigen; TCR, T-cell receptor; TRuC, T-cell receptor fusion construct.
    Figure Legend Snippet: CAR T cells use an antibody fragment (such as a single-chain variable fragment as shown here) as the binding domain—which is fused to CD3ζ and often contains additional co-stimulatory domains—that targets a cell surface protein. TRuC T cells also use an antibody fragment (such as a single-domain antibody as shown here) that targets a cell surface protein; however, this antibody fragment is fused to another part of the TCR:CD3 complex—shown here, it is fused to the CD3ε, using natural T-cell signaling pathways. FvTCR T cells replace the variable domains of the TCR with the light and heavy chains of the antibody. HiT T cells use TCR binding domains to target cell surface proteins directly and therefore use natural cell signaling pathways. TCR T cells use a TCR binding domain to specifically target a peptide presented by the HLA molecule. CAR, chimeric antigen receptor; FvTCR, variable fragment T-cell receptor; HiT, human leukocyte antigen–independent T-cell receptor; HLA, human leukocyte antigen; TCR, T-cell receptor; TRuC, T-cell receptor fusion construct.

    Techniques Used: Binding Assay, Construct

    (A) Representative example of surface plasmon resonance data. Equilibrium binding with a fit to a 1:1 Langmuir equation (one of four independent experiments is shown). (B) Epitope analysis of T-cell receptor binding to full-length (M7) and truncated (M1–6, MN1–4) mesothelin. Reduced binding is defined as a reduction in affinity (greater than five-fold increase in K D ) or reduction in maximal binding response (maximal binding response less than 10% of the theoretical maximum based on the amount of immobilized mesothelin). (C) ntd 2B9 Jurkat T cells (left column) and mesothelin HiT–transduced Jurkat T cells (right column) were analyzed for CD3 expression, demonstrating successful transduction. (D) Jurkat T cells were cultured alone or co-cultured with antigen-positive (HCT116, K562.MSLN) or antigen-negative (K562) cell lines. CD69 surface expression was monitored by flow cytometry. HiT, human leukocyte antigen–independent T-cell receptor; K D , equilibrium dissociation constant; ntd, non-transduced; T 1/2 , dissociation half-life.
    Figure Legend Snippet: (A) Representative example of surface plasmon resonance data. Equilibrium binding with a fit to a 1:1 Langmuir equation (one of four independent experiments is shown). (B) Epitope analysis of T-cell receptor binding to full-length (M7) and truncated (M1–6, MN1–4) mesothelin. Reduced binding is defined as a reduction in affinity (greater than five-fold increase in K D ) or reduction in maximal binding response (maximal binding response less than 10% of the theoretical maximum based on the amount of immobilized mesothelin). (C) ntd 2B9 Jurkat T cells (left column) and mesothelin HiT–transduced Jurkat T cells (right column) were analyzed for CD3 expression, demonstrating successful transduction. (D) Jurkat T cells were cultured alone or co-cultured with antigen-positive (HCT116, K562.MSLN) or antigen-negative (K562) cell lines. CD69 surface expression was monitored by flow cytometry. HiT, human leukocyte antigen–independent T-cell receptor; K D , equilibrium dissociation constant; ntd, non-transduced; T 1/2 , dissociation half-life.

    Techniques Used: SPR Assay, Binding Assay, Expressing, Transduction, Cell Culture, Flow Cytometry

    (A) The activity of primary T cells in either the CD4 or CD8 subset were studied for the mesothelin HiT, a mesothelin TRuC, and an HLA-A02*01 displayed MAGE-A4 peptide targeting TCR against a MAGE-A4–positive and mesothelin–over-expressing A375 cell line. Before co-culture with target cells, CD4 or CD8 T cells were incubated with anti-CD4 or anti-CD8 antibodies, respectively; after overnight incubation, IFN-γ release into the supernatants was measured using enzyme-linked immunosorbent assay. Data were analyzed using a two-way analysis of variance n.s. >0.05 * p ≤ 0.05, ** p ≤ 0.01 (GraphPad Prism). (B) Inhibition of the mesothelin-targeting HiT and TRuC by soluble mesothelin was measured using a modified Jurkat cell line expressing luciferase from an IL-2 promoter (TCR/CD3 Effector Cells IL-2, Promega). A mesothelin-positive cell line (Capan-2) and a mesothelin-negative cell line (K562) were incubated with a range of mesothelin concentrations between 0.04 and 40 μM, before being cultured with effector cells. Activation of the IL-2 promoter on effector cells was detected at 6 h. Transduction efficiencies for the transduced T cells can be found in . ctrl, control; HiT, human leukocyte antigen–independent T-cell receptor; IFN-γ, interferon γ; IL-2, interkeukin-2; n.s., non-significant; ntd, non-transduced; RLU, relative light units; TCR, T-cell receptor; TRuC, T-cell receptor fusion construct.
    Figure Legend Snippet: (A) The activity of primary T cells in either the CD4 or CD8 subset were studied for the mesothelin HiT, a mesothelin TRuC, and an HLA-A02*01 displayed MAGE-A4 peptide targeting TCR against a MAGE-A4–positive and mesothelin–over-expressing A375 cell line. Before co-culture with target cells, CD4 or CD8 T cells were incubated with anti-CD4 or anti-CD8 antibodies, respectively; after overnight incubation, IFN-γ release into the supernatants was measured using enzyme-linked immunosorbent assay. Data were analyzed using a two-way analysis of variance n.s. >0.05 * p ≤ 0.05, ** p ≤ 0.01 (GraphPad Prism). (B) Inhibition of the mesothelin-targeting HiT and TRuC by soluble mesothelin was measured using a modified Jurkat cell line expressing luciferase from an IL-2 promoter (TCR/CD3 Effector Cells IL-2, Promega). A mesothelin-positive cell line (Capan-2) and a mesothelin-negative cell line (K562) were incubated with a range of mesothelin concentrations between 0.04 and 40 μM, before being cultured with effector cells. Activation of the IL-2 promoter on effector cells was detected at 6 h. Transduction efficiencies for the transduced T cells can be found in . ctrl, control; HiT, human leukocyte antigen–independent T-cell receptor; IFN-γ, interferon γ; IL-2, interkeukin-2; n.s., non-significant; ntd, non-transduced; RLU, relative light units; TCR, T-cell receptor; TRuC, T-cell receptor fusion construct.

    Techniques Used: Activity Assay, Expressing, Co-Culture Assay, Incubation, Enzyme-linked Immunosorbent Assay, Inhibition, Modification, Luciferase, Cell Culture, Activation Assay, Transduction, Construct

    anti cd3 apc  (Thermo Fisher)


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    Thermo Fisher anti cd3 apc
    Anti Cd3 Apc, 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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    anti cd3 apc  (Thermo Fisher)


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    Thermo Fisher anti cd3 apc
    Anti Cd3 Apc, 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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    anti cd3 apc  (Thermo Fisher)


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    Thermo Fisher anti cd3 apc
    Anti Cd3 Apc, 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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    anti cd3 apc  (Thermo Fisher)


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    Thermo Fisher anti cd3 apc
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    cd3 apc h7  (Thermo Fisher)


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    Thermo Fisher cd3 apc h7
    Cd3 Apc H7, 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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    anti cd3 apc  (Thermo Fisher)


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    Thermo Fisher anti cd3 apc
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    anti cd3 apc  (Thermo Fisher)


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    Thermo Fisher anti cd3 apc
    (a) Quantification and representative images of Panc02 WT and PD-L2-KO tumors in nude mice, untreated and treated with doxorubicin on days 7 and 10. Luminescence units are photon/sec/cm 2 /stereoradian in the images. N = 3 for PD-L2-WT + PBS mice, n = 5 for PD-L2-KO + PBS, n = 6 for both doxo-treated groups. (b) Gating strategy to define circulating CD4 + and CD8 + T cell populations. (c) Percentage of CD4 + and CD8 + T cells among total CD45 + <t>CD3</t> + cells in blood of mice after treatment with blocking anti-CD4 and anti-CD8 antibodies. N = 6-7 mice. Data are presented as mean ± SEM.
    Anti Cd3 Apc, 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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    1) Product Images from "The efficacy of chemotherapy is limited by intratumoral senescent cells expressing PD-L2"

    Article Title: The efficacy of chemotherapy is limited by intratumoral senescent cells expressing PD-L2

    Journal: Nature Cancer

    doi: 10.1038/s43018-023-00712-x

    (a) Quantification and representative images of Panc02 WT and PD-L2-KO tumors in nude mice, untreated and treated with doxorubicin on days 7 and 10. Luminescence units are photon/sec/cm 2 /stereoradian in the images. N = 3 for PD-L2-WT + PBS mice, n = 5 for PD-L2-KO + PBS, n = 6 for both doxo-treated groups. (b) Gating strategy to define circulating CD4 + and CD8 + T cell populations. (c) Percentage of CD4 + and CD8 + T cells among total CD45 + CD3 + cells in blood of mice after treatment with blocking anti-CD4 and anti-CD8 antibodies. N = 6-7 mice. Data are presented as mean ± SEM.
    Figure Legend Snippet: (a) Quantification and representative images of Panc02 WT and PD-L2-KO tumors in nude mice, untreated and treated with doxorubicin on days 7 and 10. Luminescence units are photon/sec/cm 2 /stereoradian in the images. N = 3 for PD-L2-WT + PBS mice, n = 5 for PD-L2-KO + PBS, n = 6 for both doxo-treated groups. (b) Gating strategy to define circulating CD4 + and CD8 + T cell populations. (c) Percentage of CD4 + and CD8 + T cells among total CD45 + CD3 + cells in blood of mice after treatment with blocking anti-CD4 and anti-CD8 antibodies. N = 6-7 mice. Data are presented as mean ± SEM.

    Techniques Used: Blocking Assay

    (a) Percentage of NK cells (NK1.1 + ), macrophages (CD11b + Gr1 - ) and lymphocytes (CD3 + TCRb + ) relative to total CD45 + cells, in WT and PD-L2-KO tumors untreated or treated with doxorubicin at days 7 and 10, analysed by mass cytometry. N = 4 mice for all conditions. (b) Percentage of PD-1 + cells among subsets of infiltrating T cells, analysed by mass cytometry. N = 3 mice for WT + PBS and PD-L2-KO + doxo, n = 4 for WT + doxo and PD-L2-KO + PBS (c) Quantification of tumor infiltrating CD3 + lymphocytes in WT and PD-L2-KO tumors, untreated or treated with doxorubicin, analysed by immunohistochemistry. N = 4 for PBS-treated groups and n = 5 for doxo-treated groups. None of the changes were significant (1 way ANOVA, Tukey post-test). Data are presented as mean ± SEM. (d) Representative Gr1 stainings in sections of PD-L2-KO tumors treated with doxorubicin and subject to depletion of CD4 + (n = 8) or CD8 + (n = 9) T cells, as well as IgG-treated controls (n = 6) from Fig. .
    Figure Legend Snippet: (a) Percentage of NK cells (NK1.1 + ), macrophages (CD11b + Gr1 - ) and lymphocytes (CD3 + TCRb + ) relative to total CD45 + cells, in WT and PD-L2-KO tumors untreated or treated with doxorubicin at days 7 and 10, analysed by mass cytometry. N = 4 mice for all conditions. (b) Percentage of PD-1 + cells among subsets of infiltrating T cells, analysed by mass cytometry. N = 3 mice for WT + PBS and PD-L2-KO + doxo, n = 4 for WT + doxo and PD-L2-KO + PBS (c) Quantification of tumor infiltrating CD3 + lymphocytes in WT and PD-L2-KO tumors, untreated or treated with doxorubicin, analysed by immunohistochemistry. N = 4 for PBS-treated groups and n = 5 for doxo-treated groups. None of the changes were significant (1 way ANOVA, Tukey post-test). Data are presented as mean ± SEM. (d) Representative Gr1 stainings in sections of PD-L2-KO tumors treated with doxorubicin and subject to depletion of CD4 + (n = 8) or CD8 + (n = 9) T cells, as well as IgG-treated controls (n = 6) from Fig. .

    Techniques Used: Mass Cytometry, Immunohistochemistry

    a , Tumor growth in PyMT mice treated weekly (from week 9 until week 12) with αPD-L2 alone (TY25, 10 mg kg −1 ) or the same dose of isotype control every 3 days, doxorubicin alone or a combination of both as indicated. αPD-L2 monotherapy ( n = 3 mice); PBS and doxorubicin + IgG groups ( n = 4); doxorubicin + αPD-L2 ( n = 5). Two-way ANOVA. Inverted red triangles indicate day of doxorubicin treatment. b , c , SA-β-gal, CD3, CD4 and CD8 staining of untreated tumors and tumors treated with doxorubicin alone or in combination with αPD-L2 blocking antibody (TY25, 10 mg kg −1 ) or the same dose of isotype control ( b ), as in a , and analyzed at week 13 ( c ). Vehicle-treated group ( n = 4); doxorubicin-treated groups ( n = 5). Representative images are shown. Statistical significance shown versus the rest of the experimental conditions. One-way ANOVA with Tukey post-hoc test. Scale bar, 100 μm. d , SA-β-gal and PD-L2 costaining in tumor samples from PyMT mice treated weekly with doxorubicin (4 mg kg −1 ) at postnatal weeks 9–12. The indicated groups were treated with αPD-L2 (TY25, 10 mg kg −1 ), or the same dose of IgG isotype control, every 3 days. Representative images and quantification are shown. Samples were obtained at postnatal week 13 (that is, 7 days after the last doxorubicin dose). Vehicle-treated group ( n = 4 mice); doxorubicin-treated groups ( n = 5 mice). A one-way ANOVA was used for significant changes versus the rest of the conditions. Scale bar, 100 μm. e , SA-β-gal staining, p21 mRNA levels, PD-L2 mRNA levels and PD-L2 protein levels measured using fluorescence-activated cell sorting (FACS), with and without bleomycin treatment, in human head and neck cancer primary culture from patient VHIO-008. Gene expression ( n = 4 independent replicates); FACS ( n = 1). Scale bar, 50 μm. Two-sided t -tests were used.
    Figure Legend Snippet: a , Tumor growth in PyMT mice treated weekly (from week 9 until week 12) with αPD-L2 alone (TY25, 10 mg kg −1 ) or the same dose of isotype control every 3 days, doxorubicin alone or a combination of both as indicated. αPD-L2 monotherapy ( n = 3 mice); PBS and doxorubicin + IgG groups ( n = 4); doxorubicin + αPD-L2 ( n = 5). Two-way ANOVA. Inverted red triangles indicate day of doxorubicin treatment. b , c , SA-β-gal, CD3, CD4 and CD8 staining of untreated tumors and tumors treated with doxorubicin alone or in combination with αPD-L2 blocking antibody (TY25, 10 mg kg −1 ) or the same dose of isotype control ( b ), as in a , and analyzed at week 13 ( c ). Vehicle-treated group ( n = 4); doxorubicin-treated groups ( n = 5). Representative images are shown. Statistical significance shown versus the rest of the experimental conditions. One-way ANOVA with Tukey post-hoc test. Scale bar, 100 μm. d , SA-β-gal and PD-L2 costaining in tumor samples from PyMT mice treated weekly with doxorubicin (4 mg kg −1 ) at postnatal weeks 9–12. The indicated groups were treated with αPD-L2 (TY25, 10 mg kg −1 ), or the same dose of IgG isotype control, every 3 days. Representative images and quantification are shown. Samples were obtained at postnatal week 13 (that is, 7 days after the last doxorubicin dose). Vehicle-treated group ( n = 4 mice); doxorubicin-treated groups ( n = 5 mice). A one-way ANOVA was used for significant changes versus the rest of the conditions. Scale bar, 100 μm. e , SA-β-gal staining, p21 mRNA levels, PD-L2 mRNA levels and PD-L2 protein levels measured using fluorescence-activated cell sorting (FACS), with and without bleomycin treatment, in human head and neck cancer primary culture from patient VHIO-008. Gene expression ( n = 4 independent replicates); FACS ( n = 1). Scale bar, 50 μm. Two-sided t -tests were used.

    Techniques Used: Staining, Blocking Assay, Fluorescence, FACS, Expressing

    cd3 apc h7  (Thermo Fisher)


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    Thermo Fisher cd3 apc h7
    A Staining of magnetically CD8-enriched PBMCs from a representative HLA-A2+ donor #3, and HLA-A3+ donor #3. Cells were stained with a BV711-labeled multimer loaded with Flu MP 58-66 peptide (MMr), alone (first two columns, Flu-only panel)) or in combination with APC-labelled HLA-A2 and PE-labelled HLA-A3 dextramers (Dex) loaded with the indicated peptide (third and fourth columns, full panel). Each dot plot is gated on viable <t>CD3+</t> cells and indicates the frequency of Dex+ or MMr+ cells out of total CD8 + T cells. Contour plots on the right of each dot plot indicate the percent naïve (CD45RA + CCR7 + ) fraction out of the corresponding Dex+ or MMr+ fraction. B Frequency of Dex+CD8+ cells out of total CD8 + T cells for the 4 HLA-A2+ (left panel) and HLA-A3+ (right panel) donors studied. Horizontal dotted lines indicate the expected T-cell frequency range of naïve CD8 + T cells. Bars indicate median values, with median Dex+CD8+ and total CD8+ counts indicated below each distribution. C Percent naïve cells out of total Dex+CD8+ cells for the same 4 HLA-A2+ and HLA-A3+ donors. Bars indicate median values; values were excluded for one HLA-A3+ donor (#2, square symbol) for whom < 5 Dex+ cells were counted. Median naïve Dex+CD8+ counts are indicated below each distribution.
    Cd3 Apc H7, 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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    1) Product Images from "MARS an improved de novo peptide candidate selection method for non-canonical antigen target discovery in cancer"

    Article Title: MARS an improved de novo peptide candidate selection method for non-canonical antigen target discovery in cancer

    Journal: Nature Communications

    doi: 10.1038/s41467-023-44460-z

    A Staining of magnetically CD8-enriched PBMCs from a representative HLA-A2+ donor #3, and HLA-A3+ donor #3. Cells were stained with a BV711-labeled multimer loaded with Flu MP 58-66 peptide (MMr), alone (first two columns, Flu-only panel)) or in combination with APC-labelled HLA-A2 and PE-labelled HLA-A3 dextramers (Dex) loaded with the indicated peptide (third and fourth columns, full panel). Each dot plot is gated on viable CD3+ cells and indicates the frequency of Dex+ or MMr+ cells out of total CD8 + T cells. Contour plots on the right of each dot plot indicate the percent naïve (CD45RA + CCR7 + ) fraction out of the corresponding Dex+ or MMr+ fraction. B Frequency of Dex+CD8+ cells out of total CD8 + T cells for the 4 HLA-A2+ (left panel) and HLA-A3+ (right panel) donors studied. Horizontal dotted lines indicate the expected T-cell frequency range of naïve CD8 + T cells. Bars indicate median values, with median Dex+CD8+ and total CD8+ counts indicated below each distribution. C Percent naïve cells out of total Dex+CD8+ cells for the same 4 HLA-A2+ and HLA-A3+ donors. Bars indicate median values; values were excluded for one HLA-A3+ donor (#2, square symbol) for whom < 5 Dex+ cells were counted. Median naïve Dex+CD8+ counts are indicated below each distribution.
    Figure Legend Snippet: A Staining of magnetically CD8-enriched PBMCs from a representative HLA-A2+ donor #3, and HLA-A3+ donor #3. Cells were stained with a BV711-labeled multimer loaded with Flu MP 58-66 peptide (MMr), alone (first two columns, Flu-only panel)) or in combination with APC-labelled HLA-A2 and PE-labelled HLA-A3 dextramers (Dex) loaded with the indicated peptide (third and fourth columns, full panel). Each dot plot is gated on viable CD3+ cells and indicates the frequency of Dex+ or MMr+ cells out of total CD8 + T cells. Contour plots on the right of each dot plot indicate the percent naïve (CD45RA + CCR7 + ) fraction out of the corresponding Dex+ or MMr+ fraction. B Frequency of Dex+CD8+ cells out of total CD8 + T cells for the 4 HLA-A2+ (left panel) and HLA-A3+ (right panel) donors studied. Horizontal dotted lines indicate the expected T-cell frequency range of naïve CD8 + T cells. Bars indicate median values, with median Dex+CD8+ and total CD8+ counts indicated below each distribution. C Percent naïve cells out of total Dex+CD8+ cells for the same 4 HLA-A2+ and HLA-A3+ donors. Bars indicate median values; values were excluded for one HLA-A3+ donor (#2, square symbol) for whom < 5 Dex+ cells were counted. Median naïve Dex+CD8+ counts are indicated below each distribution.

    Techniques Used: Staining, Labeling

    anti human cd3 apc  (Thermo Fisher)


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    Thermo Fisher anti human cd3 apc
    Transgenic Jurkat 76 TPR cell lines co-expressing CD4 and HA1.7 or Ob.1A12 TCRs. Surface staining for the expression of CD4, <t>CD3,</t> and TCR molecules on Jurkat 76 TPR cell lines transduced with lentiviruses coding for CD4 and TCR. The analysis was carried out with flow cytometry. Values indicate the percentage of positive antigen expression. Blue-shaded histograms represent non-transduced Jurkat 76 TPR cells stained with the designated fluorophore-conjugated antibody.
    Anti Human Cd3 Apc, 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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    1) Product Images from "Genetically engineered CD80–pMHC-harboring extracellular vesicles for antigen-specific CD4 + T-cell engagement"

    Article Title: Genetically engineered CD80–pMHC-harboring extracellular vesicles for antigen-specific CD4 + T-cell engagement

    Journal: Frontiers in Bioengineering and Biotechnology

    doi: 10.3389/fbioe.2023.1341685

    Transgenic Jurkat 76 TPR cell lines co-expressing CD4 and HA1.7 or Ob.1A12 TCRs. Surface staining for the expression of CD4, CD3, and TCR molecules on Jurkat 76 TPR cell lines transduced with lentiviruses coding for CD4 and TCR. The analysis was carried out with flow cytometry. Values indicate the percentage of positive antigen expression. Blue-shaded histograms represent non-transduced Jurkat 76 TPR cells stained with the designated fluorophore-conjugated antibody.
    Figure Legend Snippet: Transgenic Jurkat 76 TPR cell lines co-expressing CD4 and HA1.7 or Ob.1A12 TCRs. Surface staining for the expression of CD4, CD3, and TCR molecules on Jurkat 76 TPR cell lines transduced with lentiviruses coding for CD4 and TCR. The analysis was carried out with flow cytometry. Values indicate the percentage of positive antigen expression. Blue-shaded histograms represent non-transduced Jurkat 76 TPR cells stained with the designated fluorophore-conjugated antibody.

    Techniques Used: Transgenic Assay, Expressing, Staining, Transduction, Flow Cytometry

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    Thermo Fisher anti cd3 apc efluor780
    CAR T cells use an antibody fragment (such as a single-chain variable fragment as shown here) as the binding domain—which is fused to CD3ζ and often contains additional co-stimulatory domains—that targets a cell surface protein. TRuC T cells also use an antibody fragment (such as a single-domain antibody as shown here) that targets a cell surface protein; however, this antibody fragment is fused to another part of the <t>TCR:CD3</t> complex—shown here, it is fused to the CD3ε, using natural T-cell signaling pathways. FvTCR T cells replace the variable domains of the TCR with the light and heavy chains of the antibody. HiT T cells use TCR binding domains to target cell surface proteins directly and therefore use natural cell signaling pathways. TCR T cells use a TCR binding domain to specifically target a peptide presented by the HLA molecule. CAR, chimeric antigen receptor; FvTCR, variable fragment T-cell receptor; HiT, human leukocyte antigen–independent T-cell receptor; HLA, human leukocyte antigen; TCR, T-cell receptor; TRuC, T-cell receptor fusion construct.
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    CAR T cells use an antibody fragment (such as a single-chain variable fragment as shown here) as the binding domain—which is fused to CD3ζ and often contains additional co-stimulatory domains—that targets a cell surface protein. TRuC T cells also use an antibody fragment (such as a single-domain antibody as shown here) that targets a cell surface protein; however, this antibody fragment is fused to another part of the <t>TCR:CD3</t> complex—shown here, it is fused to the CD3ε, using natural T-cell signaling pathways. FvTCR T cells replace the variable domains of the TCR with the light and heavy chains of the antibody. HiT T cells use TCR binding domains to target cell surface proteins directly and therefore use natural cell signaling pathways. TCR T cells use a TCR binding domain to specifically target a peptide presented by the HLA molecule. CAR, chimeric antigen receptor; FvTCR, variable fragment T-cell receptor; HiT, human leukocyte antigen–independent T-cell receptor; HLA, human leukocyte antigen; TCR, T-cell receptor; TRuC, T-cell receptor fusion construct.
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    CAR T cells use an antibody fragment (such as a single-chain variable fragment as shown here) as the binding domain—which is fused to CD3ζ and often contains additional co-stimulatory domains—that targets a cell surface protein. TRuC T cells also use an antibody fragment (such as a single-domain antibody as shown here) that targets a cell surface protein; however, this antibody fragment is fused to another part of the <t>TCR:CD3</t> complex—shown here, it is fused to the CD3ε, using natural T-cell signaling pathways. FvTCR T cells replace the variable domains of the TCR with the light and heavy chains of the antibody. HiT T cells use TCR binding domains to target cell surface proteins directly and therefore use natural cell signaling pathways. TCR T cells use a TCR binding domain to specifically target a peptide presented by the HLA molecule. CAR, chimeric antigen receptor; FvTCR, variable fragment T-cell receptor; HiT, human leukocyte antigen–independent T-cell receptor; HLA, human leukocyte antigen; TCR, T-cell receptor; TRuC, T-cell receptor fusion construct.
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    Transgenic Jurkat 76 TPR cell lines co-expressing CD4 and HA1.7 or Ob.1A12 TCRs. Surface staining for the expression of CD4, <t>CD3,</t> and TCR molecules on Jurkat 76 TPR cell lines transduced with lentiviruses coding for CD4 and TCR. The analysis was carried out with flow cytometry. Values indicate the percentage of positive antigen expression. Blue-shaded histograms represent non-transduced Jurkat 76 TPR cells stained with the designated fluorophore-conjugated antibody.
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    CAR T cells use an antibody fragment (such as a single-chain variable fragment as shown here) as the binding domain—which is fused to CD3ζ and often contains additional co-stimulatory domains—that targets a cell surface protein. TRuC T cells also use an antibody fragment (such as a single-domain antibody as shown here) that targets a cell surface protein; however, this antibody fragment is fused to another part of the TCR:CD3 complex—shown here, it is fused to the CD3ε, using natural T-cell signaling pathways. FvTCR T cells replace the variable domains of the TCR with the light and heavy chains of the antibody. HiT T cells use TCR binding domains to target cell surface proteins directly and therefore use natural cell signaling pathways. TCR T cells use a TCR binding domain to specifically target a peptide presented by the HLA molecule. CAR, chimeric antigen receptor; FvTCR, variable fragment T-cell receptor; HiT, human leukocyte antigen–independent T-cell receptor; HLA, human leukocyte antigen; TCR, T-cell receptor; TRuC, T-cell receptor fusion construct.

    Journal: PLOS ONE

    Article Title: Selection, engineering, and in vivo testing of a human leukocyte antigen–independent T-cell receptor recognizing human mesothelin

    doi: 10.1371/journal.pone.0301175

    Figure Lengend Snippet: CAR T cells use an antibody fragment (such as a single-chain variable fragment as shown here) as the binding domain—which is fused to CD3ζ and often contains additional co-stimulatory domains—that targets a cell surface protein. TRuC T cells also use an antibody fragment (such as a single-domain antibody as shown here) that targets a cell surface protein; however, this antibody fragment is fused to another part of the TCR:CD3 complex—shown here, it is fused to the CD3ε, using natural T-cell signaling pathways. FvTCR T cells replace the variable domains of the TCR with the light and heavy chains of the antibody. HiT T cells use TCR binding domains to target cell surface proteins directly and therefore use natural cell signaling pathways. TCR T cells use a TCR binding domain to specifically target a peptide presented by the HLA molecule. CAR, chimeric antigen receptor; FvTCR, variable fragment T-cell receptor; HiT, human leukocyte antigen–independent T-cell receptor; HLA, human leukocyte antigen; TCR, T-cell receptor; TRuC, T-cell receptor fusion construct.

    Article Snippet: Cells were washed once with PBS with 0.1% fetal bovine serum (Gibco) before incubation with anti-CD69-PE (12-0699-42, Thermo Fisher Scientific), anti-CD3-APC-eFluor780 (47-0038-42, Thermo Fisher Scientific), and anti-CD8-VioBlue (130-110-684, Miltenyi Biotec) for 30 min at 4°C.

    Techniques: Binding Assay, Construct

    (A) Representative example of surface plasmon resonance data. Equilibrium binding with a fit to a 1:1 Langmuir equation (one of four independent experiments is shown). (B) Epitope analysis of T-cell receptor binding to full-length (M7) and truncated (M1–6, MN1–4) mesothelin. Reduced binding is defined as a reduction in affinity (greater than five-fold increase in K D ) or reduction in maximal binding response (maximal binding response less than 10% of the theoretical maximum based on the amount of immobilized mesothelin). (C) ntd 2B9 Jurkat T cells (left column) and mesothelin HiT–transduced Jurkat T cells (right column) were analyzed for CD3 expression, demonstrating successful transduction. (D) Jurkat T cells were cultured alone or co-cultured with antigen-positive (HCT116, K562.MSLN) or antigen-negative (K562) cell lines. CD69 surface expression was monitored by flow cytometry. HiT, human leukocyte antigen–independent T-cell receptor; K D , equilibrium dissociation constant; ntd, non-transduced; T 1/2 , dissociation half-life.

    Journal: PLOS ONE

    Article Title: Selection, engineering, and in vivo testing of a human leukocyte antigen–independent T-cell receptor recognizing human mesothelin

    doi: 10.1371/journal.pone.0301175

    Figure Lengend Snippet: (A) Representative example of surface plasmon resonance data. Equilibrium binding with a fit to a 1:1 Langmuir equation (one of four independent experiments is shown). (B) Epitope analysis of T-cell receptor binding to full-length (M7) and truncated (M1–6, MN1–4) mesothelin. Reduced binding is defined as a reduction in affinity (greater than five-fold increase in K D ) or reduction in maximal binding response (maximal binding response less than 10% of the theoretical maximum based on the amount of immobilized mesothelin). (C) ntd 2B9 Jurkat T cells (left column) and mesothelin HiT–transduced Jurkat T cells (right column) were analyzed for CD3 expression, demonstrating successful transduction. (D) Jurkat T cells were cultured alone or co-cultured with antigen-positive (HCT116, K562.MSLN) or antigen-negative (K562) cell lines. CD69 surface expression was monitored by flow cytometry. HiT, human leukocyte antigen–independent T-cell receptor; K D , equilibrium dissociation constant; ntd, non-transduced; T 1/2 , dissociation half-life.

    Article Snippet: Cells were washed once with PBS with 0.1% fetal bovine serum (Gibco) before incubation with anti-CD69-PE (12-0699-42, Thermo Fisher Scientific), anti-CD3-APC-eFluor780 (47-0038-42, Thermo Fisher Scientific), and anti-CD8-VioBlue (130-110-684, Miltenyi Biotec) for 30 min at 4°C.

    Techniques: SPR Assay, Binding Assay, Expressing, Transduction, Cell Culture, Flow Cytometry

    (A) The activity of primary T cells in either the CD4 or CD8 subset were studied for the mesothelin HiT, a mesothelin TRuC, and an HLA-A02*01 displayed MAGE-A4 peptide targeting TCR against a MAGE-A4–positive and mesothelin–over-expressing A375 cell line. Before co-culture with target cells, CD4 or CD8 T cells were incubated with anti-CD4 or anti-CD8 antibodies, respectively; after overnight incubation, IFN-γ release into the supernatants was measured using enzyme-linked immunosorbent assay. Data were analyzed using a two-way analysis of variance n.s. >0.05 * p ≤ 0.05, ** p ≤ 0.01 (GraphPad Prism). (B) Inhibition of the mesothelin-targeting HiT and TRuC by soluble mesothelin was measured using a modified Jurkat cell line expressing luciferase from an IL-2 promoter (TCR/CD3 Effector Cells IL-2, Promega). A mesothelin-positive cell line (Capan-2) and a mesothelin-negative cell line (K562) were incubated with a range of mesothelin concentrations between 0.04 and 40 μM, before being cultured with effector cells. Activation of the IL-2 promoter on effector cells was detected at 6 h. Transduction efficiencies for the transduced T cells can be found in . ctrl, control; HiT, human leukocyte antigen–independent T-cell receptor; IFN-γ, interferon γ; IL-2, interkeukin-2; n.s., non-significant; ntd, non-transduced; RLU, relative light units; TCR, T-cell receptor; TRuC, T-cell receptor fusion construct.

    Journal: PLOS ONE

    Article Title: Selection, engineering, and in vivo testing of a human leukocyte antigen–independent T-cell receptor recognizing human mesothelin

    doi: 10.1371/journal.pone.0301175

    Figure Lengend Snippet: (A) The activity of primary T cells in either the CD4 or CD8 subset were studied for the mesothelin HiT, a mesothelin TRuC, and an HLA-A02*01 displayed MAGE-A4 peptide targeting TCR against a MAGE-A4–positive and mesothelin–over-expressing A375 cell line. Before co-culture with target cells, CD4 or CD8 T cells were incubated with anti-CD4 or anti-CD8 antibodies, respectively; after overnight incubation, IFN-γ release into the supernatants was measured using enzyme-linked immunosorbent assay. Data were analyzed using a two-way analysis of variance n.s. >0.05 * p ≤ 0.05, ** p ≤ 0.01 (GraphPad Prism). (B) Inhibition of the mesothelin-targeting HiT and TRuC by soluble mesothelin was measured using a modified Jurkat cell line expressing luciferase from an IL-2 promoter (TCR/CD3 Effector Cells IL-2, Promega). A mesothelin-positive cell line (Capan-2) and a mesothelin-negative cell line (K562) were incubated with a range of mesothelin concentrations between 0.04 and 40 μM, before being cultured with effector cells. Activation of the IL-2 promoter on effector cells was detected at 6 h. Transduction efficiencies for the transduced T cells can be found in . ctrl, control; HiT, human leukocyte antigen–independent T-cell receptor; IFN-γ, interferon γ; IL-2, interkeukin-2; n.s., non-significant; ntd, non-transduced; RLU, relative light units; TCR, T-cell receptor; TRuC, T-cell receptor fusion construct.

    Article Snippet: Cells were washed once with PBS with 0.1% fetal bovine serum (Gibco) before incubation with anti-CD69-PE (12-0699-42, Thermo Fisher Scientific), anti-CD3-APC-eFluor780 (47-0038-42, Thermo Fisher Scientific), and anti-CD8-VioBlue (130-110-684, Miltenyi Biotec) for 30 min at 4°C.

    Techniques: Activity Assay, Expressing, Co-Culture Assay, Incubation, Enzyme-linked Immunosorbent Assay, Inhibition, Modification, Luciferase, Cell Culture, Activation Assay, Transduction, Construct

    Transgenic Jurkat 76 TPR cell lines co-expressing CD4 and HA1.7 or Ob.1A12 TCRs. Surface staining for the expression of CD4, CD3, and TCR molecules on Jurkat 76 TPR cell lines transduced with lentiviruses coding for CD4 and TCR. The analysis was carried out with flow cytometry. Values indicate the percentage of positive antigen expression. Blue-shaded histograms represent non-transduced Jurkat 76 TPR cells stained with the designated fluorophore-conjugated antibody.

    Journal: Frontiers in Bioengineering and Biotechnology

    Article Title: Genetically engineered CD80–pMHC-harboring extracellular vesicles for antigen-specific CD4 + T-cell engagement

    doi: 10.3389/fbioe.2023.1341685

    Figure Lengend Snippet: Transgenic Jurkat 76 TPR cell lines co-expressing CD4 and HA1.7 or Ob.1A12 TCRs. Surface staining for the expression of CD4, CD3, and TCR molecules on Jurkat 76 TPR cell lines transduced with lentiviruses coding for CD4 and TCR. The analysis was carried out with flow cytometry. Values indicate the percentage of positive antigen expression. Blue-shaded histograms represent non-transduced Jurkat 76 TPR cells stained with the designated fluorophore-conjugated antibody.

    Article Snippet: The following fluorophore-conjugated antibodies were used: anti-human CD80-PE (clone W17149D, BioLegend), anti-human HLA-DR-APC (clone L243, BioLegend), anti-human CD4-APC-Alexa Fluor 750 (clone S3.5, Thermo Fisher Scientific), anti-human CD3-APC or CD3-FITC (clone OKT3, BioLegend), anti-human TCR α/β-APC (clone IP26, BioLegend), anti-human CD69-APC (clone FN50, BioLegend), anti-human IFNγ-PE (clone 4S.B3, BioLegend), and anti-human CD11c-FITC (clone 3.9, BioLegend).

    Techniques: Transgenic Assay, Expressing, Staining, Transduction, Flow Cytometry