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5t4 specific mab  (R&D Systems)


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    R&D Systems 5t4 specific mab
    High throughput bulk sequencing of the T-cell receptor β chain for 5T4 p17 -specific CD8 + T-cell clones isolated from healthy or kidney cancer donors
    5t4 Specific Mab, supplied by R&D Systems, used in various techniques. Bioz Stars score: 94/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/5t4 specific mab/product/R&D Systems
    Average 94 stars, based on 7 article reviews
    5t4 specific mab - by Bioz Stars, 2026-02
    94/100 stars

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    1) Product Images from "Preclinical development of T-cell receptor-engineered T-cell therapy targeting the 5T4 tumor antigen on renal cell carcinoma"

    Article Title: Preclinical development of T-cell receptor-engineered T-cell therapy targeting the 5T4 tumor antigen on renal cell carcinoma

    Journal: Cancer Immunology, Immunotherapy

    doi: 10.1007/s00262-019-02419-4

    High throughput bulk sequencing of the T-cell receptor β chain for 5T4 p17 -specific CD8 + T-cell clones isolated from healthy or kidney cancer donors
    Figure Legend Snippet: High throughput bulk sequencing of the T-cell receptor β chain for 5T4 p17 -specific CD8 + T-cell clones isolated from healthy or kidney cancer donors

    Techniques Used: High Throughput Screening Assay, Sequencing, Clone Assay, Isolation

    Characteristics of 5T4 p17 -specific CDR3 regions of TRA and TRB . IMGT junction analysis is shown for a TRA -CDR3 regions and b TRB -CDR3 regions. Red boxes indicate N nucleotides and/or D regions ( TRB ) between V gene and J gene encoded sequence. c V- and J-gene usage for TRA and TRB are shown. Color shading identifies common gene use between TCRs. d The net charge for CDR1, 2, and 3 amino acid sequences from 5T4 17–25 -specific TCRs are shown
    Figure Legend Snippet: Characteristics of 5T4 p17 -specific CDR3 regions of TRA and TRB . IMGT junction analysis is shown for a TRA -CDR3 regions and b TRB -CDR3 regions. Red boxes indicate N nucleotides and/or D regions ( TRB ) between V gene and J gene encoded sequence. c V- and J-gene usage for TRA and TRB are shown. Color shading identifies common gene use between TCRs. d The net charge for CDR1, 2, and 3 amino acid sequences from 5T4 17–25 -specific TCRs are shown

    Techniques Used: Sequencing

    5T4 p17 -specific TCRs expression on CD8 + T-cells from healthy donors and redirected effector functions against peptide-pulsed T2 targets. a Vector structure of 5T4 p17 -specific TCR assembly: TRB and TRA are transcribed under control of the MSCV promotor; P2A cleaves the primary transcripts to equal molar quantity of TRA and TRB in the cytosol. The enlarged graph indicates the gene segments and CDR3s, contributing to TRA and TRB sequence assembly. The relative location for the introduced transmembrane cysteines in TRAC and TRBC are indicated. b Cell surface expression of 5T4 p17 -specific TCRs stained by 5T4 p17 /HLA-A2 tetramer at day 7 post-transduction is shown in comparison to tetramer staining of the native T-cell clone expressing the corresponding TCR (upper panels). Cell surface expression of 5T4 p17 -specific TCRs on healthy donor T-cells is shown after sorting for 5T4 p17 /HLA-A2 tetramer + cells and 12 days expansion (lower panels). CD8 + T-cells expressing 5T4 p17 -specific TCRs were tested for recognition of c T2 cells pulsed with 10 nM of 5T4 p17 or control HLA-A2-binding peptides DDX3Y 428–436 or UTY 148–156 in a 4-h cytotoxicity assay. The effector: target ratio (E:T) was 10:1. d TNF-α release was measured by ELISA in culture supernatants harvested after 18 h co-culture of effector T-cells with T2 cells pulsed with 10 nM of 5T4 p17 or the control peptide DDX3Y 428–436 at 10:1 E:T. e Effector T-cells were tested for recognition of T2 cells pulsed with 5T4p17 peptide in a 4-h cytotoxicity assay at 10:1 E:T. f TNF-α release was measured by ELISA in culture supernatants harvested after 18 h co-culture of effector T-cells with T2 cells pulsed with 5T4 p17 peptide at 10:1 E:T
    Figure Legend Snippet: 5T4 p17 -specific TCRs expression on CD8 + T-cells from healthy donors and redirected effector functions against peptide-pulsed T2 targets. a Vector structure of 5T4 p17 -specific TCR assembly: TRB and TRA are transcribed under control of the MSCV promotor; P2A cleaves the primary transcripts to equal molar quantity of TRA and TRB in the cytosol. The enlarged graph indicates the gene segments and CDR3s, contributing to TRA and TRB sequence assembly. The relative location for the introduced transmembrane cysteines in TRAC and TRBC are indicated. b Cell surface expression of 5T4 p17 -specific TCRs stained by 5T4 p17 /HLA-A2 tetramer at day 7 post-transduction is shown in comparison to tetramer staining of the native T-cell clone expressing the corresponding TCR (upper panels). Cell surface expression of 5T4 p17 -specific TCRs on healthy donor T-cells is shown after sorting for 5T4 p17 /HLA-A2 tetramer + cells and 12 days expansion (lower panels). CD8 + T-cells expressing 5T4 p17 -specific TCRs were tested for recognition of c T2 cells pulsed with 10 nM of 5T4 p17 or control HLA-A2-binding peptides DDX3Y 428–436 or UTY 148–156 in a 4-h cytotoxicity assay. The effector: target ratio (E:T) was 10:1. d TNF-α release was measured by ELISA in culture supernatants harvested after 18 h co-culture of effector T-cells with T2 cells pulsed with 10 nM of 5T4 p17 or the control peptide DDX3Y 428–436 at 10:1 E:T. e Effector T-cells were tested for recognition of T2 cells pulsed with 5T4p17 peptide in a 4-h cytotoxicity assay at 10:1 E:T. f TNF-α release was measured by ELISA in culture supernatants harvested after 18 h co-culture of effector T-cells with T2 cells pulsed with 5T4 p17 peptide at 10:1 E:T

    Techniques Used: Expressing, Plasmid Preparation, Control, Sequencing, Staining, Transduction, Comparison, Binding Assay, Cytotoxicity Assay, Enzyme-linked Immunosorbent Assay, Co-Culture Assay

    Cytotoxicity of 5T4 p17 -specific TCR transduced CD8 + T-cells against T2 pulsed with alanine-substituted 5T4 p17 peptides. a Binding of alanine-substituted 5T4 p17 peptides to HLA-A2 was determined by stabilization of HLA-A2 on the surface of peptide-pulsed T2 cells. CMV pp65 is an HLA-A2 + T-cell epitope from CMV and serves as a positive control. Cell surface HLA-A2 was assessed by immunostaining and flow cytometry. b CD8 + T-cells expressing 5T4 p17 -specific TCRs were tested for recognition of T2 cells pulsed with the progenitor 5T4 p17 peptide or HLA-A2 binding alanine-substituted variants in a 4-h cytotoxicity assay with a 10:1 E:T
    Figure Legend Snippet: Cytotoxicity of 5T4 p17 -specific TCR transduced CD8 + T-cells against T2 pulsed with alanine-substituted 5T4 p17 peptides. a Binding of alanine-substituted 5T4 p17 peptides to HLA-A2 was determined by stabilization of HLA-A2 on the surface of peptide-pulsed T2 cells. CMV pp65 is an HLA-A2 + T-cell epitope from CMV and serves as a positive control. Cell surface HLA-A2 was assessed by immunostaining and flow cytometry. b CD8 + T-cells expressing 5T4 p17 -specific TCRs were tested for recognition of T2 cells pulsed with the progenitor 5T4 p17 peptide or HLA-A2 binding alanine-substituted variants in a 4-h cytotoxicity assay with a 10:1 E:T

    Techniques Used: Binding Assay, Positive Control, Immunostaining, Flow Cytometry, Expressing, Cytotoxicity Assay

    Cytotoxicity of 5T4 p17 -specific TCR-transduced CD8 + T-cells against 5T4-expressing tumor targets. CD8 + T-cells expressing 5T4 17–25 -specific TCRs were tested for recognition of tumor target lines in a 4-h cytotoxicity assay with 10:1 E:T. For each target cell-line, 5T4 17–25 -specific TCR transduced effector CD8 + T-cells were plotted in the following order: HD_A-2, HD_A-15, HD_B-19, HD_B-21, HD_C-3, HD_C-17, KCD_D-6, and untransduced. a Targets are 5T4 + /HLA-A2 + RCC cell lines (A498, BB65, TREP, DOBSKI; red bars), the 5T4 + /HLA-A2 − RCC line (SST548; open bars) and the 5T4 − /HLA-A2 + LCL (BB65-LCL; gray shading). b Targets are 5T4 + /HLA-A2 + breast cancer line (MDA231; blue bars), a 5T4 + /HLA-A2 − breast cancer line (BT20; open bars) and 5T4 + /HLA-A2 + colon cancer line (SW480, green bars). CD8 + T-cells with (gTRAC_KO) or without (gCtrl) native TRA disruption expressing 5T4 p17 -specific TCRs were tested for recognition of c 5T4 + /HLA-A2 + RCC cell-lines (A498, BB65, DOBSKI), the 5T4 + /HLA-A2 − RCC line (SST548), the 5T4 − /HLA-A2 + BB65-LCL, and d paired primary 5T4 + /HLA-A2 + RCC and autologous PTEC cell-lines from four RCC patients
    Figure Legend Snippet: Cytotoxicity of 5T4 p17 -specific TCR-transduced CD8 + T-cells against 5T4-expressing tumor targets. CD8 + T-cells expressing 5T4 17–25 -specific TCRs were tested for recognition of tumor target lines in a 4-h cytotoxicity assay with 10:1 E:T. For each target cell-line, 5T4 17–25 -specific TCR transduced effector CD8 + T-cells were plotted in the following order: HD_A-2, HD_A-15, HD_B-19, HD_B-21, HD_C-3, HD_C-17, KCD_D-6, and untransduced. a Targets are 5T4 + /HLA-A2 + RCC cell lines (A498, BB65, TREP, DOBSKI; red bars), the 5T4 + /HLA-A2 − RCC line (SST548; open bars) and the 5T4 − /HLA-A2 + LCL (BB65-LCL; gray shading). b Targets are 5T4 + /HLA-A2 + breast cancer line (MDA231; blue bars), a 5T4 + /HLA-A2 − breast cancer line (BT20; open bars) and 5T4 + /HLA-A2 + colon cancer line (SW480, green bars). CD8 + T-cells with (gTRAC_KO) or without (gCtrl) native TRA disruption expressing 5T4 p17 -specific TCRs were tested for recognition of c 5T4 + /HLA-A2 + RCC cell-lines (A498, BB65, DOBSKI), the 5T4 + /HLA-A2 − RCC line (SST548), the 5T4 − /HLA-A2 + BB65-LCL, and d paired primary 5T4 + /HLA-A2 + RCC and autologous PTEC cell-lines from four RCC patients

    Techniques Used: Expressing, Cytotoxicity Assay, Disruption

    Cytotoxicity of 5T4 p17 -specific TCR-transduced CD8 + T-cells against MVA-5T4 infected targets. a Schematic of human 5T4 protein, p17–25 is in the signal sequence region. Trans: transmembrane region, Cyto: cytoplasmic region. b T2 cells and BB65-LCL were infected with MVA-WT or MVA-5T4, and cell surface expression of 5T4 was analyzed by immunostaining and flow cytometry 24 h after infection. The fraction of cells positive for cell surface 5T4 after MVA-5T4 infection is indicated. CD8 + T-cells expressing 5T4 p17 -specific TCRs were tested for recognition of c T2 and d BB65-LCL cells infected by MVA-WT, MVA-5T4, or uninfected cells pulsed with 10 nM 5T4 p17 peptide in a 6-h cytotoxicity assay at a 10:1 E:T
    Figure Legend Snippet: Cytotoxicity of 5T4 p17 -specific TCR-transduced CD8 + T-cells against MVA-5T4 infected targets. a Schematic of human 5T4 protein, p17–25 is in the signal sequence region. Trans: transmembrane region, Cyto: cytoplasmic region. b T2 cells and BB65-LCL were infected with MVA-WT or MVA-5T4, and cell surface expression of 5T4 was analyzed by immunostaining and flow cytometry 24 h after infection. The fraction of cells positive for cell surface 5T4 after MVA-5T4 infection is indicated. CD8 + T-cells expressing 5T4 p17 -specific TCRs were tested for recognition of c T2 and d BB65-LCL cells infected by MVA-WT, MVA-5T4, or uninfected cells pulsed with 10 nM 5T4 p17 peptide in a 6-h cytotoxicity assay at a 10:1 E:T

    Techniques Used: Infection, Sequencing, Expressing, Immunostaining, Flow Cytometry, Cytotoxicity Assay



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    5t4 specific mab  (R&D Systems)
    94
    R&D Systems 5t4 specific mab
    High throughput bulk sequencing of the T-cell receptor β chain for 5T4 p17 -specific CD8 + T-cell clones isolated from healthy or kidney cancer donors
    5t4 Specific Mab, supplied by R&D Systems, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/5t4 specific mab/product/R&D Systems
    Average 94 stars, based on 1 article reviews
    5t4 specific mab - by Bioz Stars, 2026-02
    94/100 stars
    		  Buy from Supplier

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    High throughput bulk sequencing of the T-cell receptor β chain for 5T4 p17 -specific CD8 + T-cell clones isolated from healthy or kidney cancer donors

    Journal: Cancer Immunology, Immunotherapy

    Article Title: Preclinical development of T-cell receptor-engineered T-cell therapy targeting the 5T4 tumor antigen on renal cell carcinoma

    doi: 10.1007/s00262-019-02419-4

    Figure Lengend Snippet: High throughput bulk sequencing of the T-cell receptor β chain for 5T4 p17 -specific CD8 + T-cell clones isolated from healthy or kidney cancer donors

    Article Snippet: To confirm 5T4 expression on surface after MVA-5T4 infection, target cells were stained with a 5T4-specific mAb (clone 524744; R&D systems, Minneapolis, MN) at 1 μg/mL followed by a 1:50 dilution of a secondary PE-labeled anti-mouse IgG1 mAb (clone A85-1; BD Biosciences) for analysis by flow cytometry.

    Techniques: High Throughput Screening Assay, Sequencing, Clone Assay, Isolation

    Characteristics of 5T4 p17 -specific CDR3 regions of TRA and TRB . IMGT junction analysis is shown for a TRA -CDR3 regions and b TRB -CDR3 regions. Red boxes indicate N nucleotides and/or D regions ( TRB ) between V gene and J gene encoded sequence. c V- and J-gene usage for TRA and TRB are shown. Color shading identifies common gene use between TCRs. d The net charge for CDR1, 2, and 3 amino acid sequences from 5T4 17–25 -specific TCRs are shown

    Journal: Cancer Immunology, Immunotherapy

    Article Title: Preclinical development of T-cell receptor-engineered T-cell therapy targeting the 5T4 tumor antigen on renal cell carcinoma

    doi: 10.1007/s00262-019-02419-4

    Figure Lengend Snippet: Characteristics of 5T4 p17 -specific CDR3 regions of TRA and TRB . IMGT junction analysis is shown for a TRA -CDR3 regions and b TRB -CDR3 regions. Red boxes indicate N nucleotides and/or D regions ( TRB ) between V gene and J gene encoded sequence. c V- and J-gene usage for TRA and TRB are shown. Color shading identifies common gene use between TCRs. d The net charge for CDR1, 2, and 3 amino acid sequences from 5T4 17–25 -specific TCRs are shown

    Article Snippet: To confirm 5T4 expression on surface after MVA-5T4 infection, target cells were stained with a 5T4-specific mAb (clone 524744; R&D systems, Minneapolis, MN) at 1 μg/mL followed by a 1:50 dilution of a secondary PE-labeled anti-mouse IgG1 mAb (clone A85-1; BD Biosciences) for analysis by flow cytometry.

    Techniques: Sequencing

    5T4 p17 -specific TCRs expression on CD8 + T-cells from healthy donors and redirected effector functions against peptide-pulsed T2 targets. a Vector structure of 5T4 p17 -specific TCR assembly: TRB and TRA are transcribed under control of the MSCV promotor; P2A cleaves the primary transcripts to equal molar quantity of TRA and TRB in the cytosol. The enlarged graph indicates the gene segments and CDR3s, contributing to TRA and TRB sequence assembly. The relative location for the introduced transmembrane cysteines in TRAC and TRBC are indicated. b Cell surface expression of 5T4 p17 -specific TCRs stained by 5T4 p17 /HLA-A2 tetramer at day 7 post-transduction is shown in comparison to tetramer staining of the native T-cell clone expressing the corresponding TCR (upper panels). Cell surface expression of 5T4 p17 -specific TCRs on healthy donor T-cells is shown after sorting for 5T4 p17 /HLA-A2 tetramer + cells and 12 days expansion (lower panels). CD8 + T-cells expressing 5T4 p17 -specific TCRs were tested for recognition of c T2 cells pulsed with 10 nM of 5T4 p17 or control HLA-A2-binding peptides DDX3Y 428–436 or UTY 148–156 in a 4-h cytotoxicity assay. The effector: target ratio (E:T) was 10:1. d TNF-α release was measured by ELISA in culture supernatants harvested after 18 h co-culture of effector T-cells with T2 cells pulsed with 10 nM of 5T4 p17 or the control peptide DDX3Y 428–436 at 10:1 E:T. e Effector T-cells were tested for recognition of T2 cells pulsed with 5T4p17 peptide in a 4-h cytotoxicity assay at 10:1 E:T. f TNF-α release was measured by ELISA in culture supernatants harvested after 18 h co-culture of effector T-cells with T2 cells pulsed with 5T4 p17 peptide at 10:1 E:T

    Journal: Cancer Immunology, Immunotherapy

    Article Title: Preclinical development of T-cell receptor-engineered T-cell therapy targeting the 5T4 tumor antigen on renal cell carcinoma

    doi: 10.1007/s00262-019-02419-4

    Figure Lengend Snippet: 5T4 p17 -specific TCRs expression on CD8 + T-cells from healthy donors and redirected effector functions against peptide-pulsed T2 targets. a Vector structure of 5T4 p17 -specific TCR assembly: TRB and TRA are transcribed under control of the MSCV promotor; P2A cleaves the primary transcripts to equal molar quantity of TRA and TRB in the cytosol. The enlarged graph indicates the gene segments and CDR3s, contributing to TRA and TRB sequence assembly. The relative location for the introduced transmembrane cysteines in TRAC and TRBC are indicated. b Cell surface expression of 5T4 p17 -specific TCRs stained by 5T4 p17 /HLA-A2 tetramer at day 7 post-transduction is shown in comparison to tetramer staining of the native T-cell clone expressing the corresponding TCR (upper panels). Cell surface expression of 5T4 p17 -specific TCRs on healthy donor T-cells is shown after sorting for 5T4 p17 /HLA-A2 tetramer + cells and 12 days expansion (lower panels). CD8 + T-cells expressing 5T4 p17 -specific TCRs were tested for recognition of c T2 cells pulsed with 10 nM of 5T4 p17 or control HLA-A2-binding peptides DDX3Y 428–436 or UTY 148–156 in a 4-h cytotoxicity assay. The effector: target ratio (E:T) was 10:1. d TNF-α release was measured by ELISA in culture supernatants harvested after 18 h co-culture of effector T-cells with T2 cells pulsed with 10 nM of 5T4 p17 or the control peptide DDX3Y 428–436 at 10:1 E:T. e Effector T-cells were tested for recognition of T2 cells pulsed with 5T4p17 peptide in a 4-h cytotoxicity assay at 10:1 E:T. f TNF-α release was measured by ELISA in culture supernatants harvested after 18 h co-culture of effector T-cells with T2 cells pulsed with 5T4 p17 peptide at 10:1 E:T

    Article Snippet: To confirm 5T4 expression on surface after MVA-5T4 infection, target cells were stained with a 5T4-specific mAb (clone 524744; R&D systems, Minneapolis, MN) at 1 μg/mL followed by a 1:50 dilution of a secondary PE-labeled anti-mouse IgG1 mAb (clone A85-1; BD Biosciences) for analysis by flow cytometry.

    Techniques: Expressing, Plasmid Preparation, Control, Sequencing, Staining, Transduction, Comparison, Binding Assay, Cytotoxicity Assay, Enzyme-linked Immunosorbent Assay, Co-Culture Assay

    Cytotoxicity of 5T4 p17 -specific TCR transduced CD8 + T-cells against T2 pulsed with alanine-substituted 5T4 p17 peptides. a Binding of alanine-substituted 5T4 p17 peptides to HLA-A2 was determined by stabilization of HLA-A2 on the surface of peptide-pulsed T2 cells. CMV pp65 is an HLA-A2 + T-cell epitope from CMV and serves as a positive control. Cell surface HLA-A2 was assessed by immunostaining and flow cytometry. b CD8 + T-cells expressing 5T4 p17 -specific TCRs were tested for recognition of T2 cells pulsed with the progenitor 5T4 p17 peptide or HLA-A2 binding alanine-substituted variants in a 4-h cytotoxicity assay with a 10:1 E:T

    Journal: Cancer Immunology, Immunotherapy

    Article Title: Preclinical development of T-cell receptor-engineered T-cell therapy targeting the 5T4 tumor antigen on renal cell carcinoma

    doi: 10.1007/s00262-019-02419-4

    Figure Lengend Snippet: Cytotoxicity of 5T4 p17 -specific TCR transduced CD8 + T-cells against T2 pulsed with alanine-substituted 5T4 p17 peptides. a Binding of alanine-substituted 5T4 p17 peptides to HLA-A2 was determined by stabilization of HLA-A2 on the surface of peptide-pulsed T2 cells. CMV pp65 is an HLA-A2 + T-cell epitope from CMV and serves as a positive control. Cell surface HLA-A2 was assessed by immunostaining and flow cytometry. b CD8 + T-cells expressing 5T4 p17 -specific TCRs were tested for recognition of T2 cells pulsed with the progenitor 5T4 p17 peptide or HLA-A2 binding alanine-substituted variants in a 4-h cytotoxicity assay with a 10:1 E:T

    Article Snippet: To confirm 5T4 expression on surface after MVA-5T4 infection, target cells were stained with a 5T4-specific mAb (clone 524744; R&D systems, Minneapolis, MN) at 1 μg/mL followed by a 1:50 dilution of a secondary PE-labeled anti-mouse IgG1 mAb (clone A85-1; BD Biosciences) for analysis by flow cytometry.

    Techniques: Binding Assay, Positive Control, Immunostaining, Flow Cytometry, Expressing, Cytotoxicity Assay

    Cytotoxicity of 5T4 p17 -specific TCR-transduced CD8 + T-cells against 5T4-expressing tumor targets. CD8 + T-cells expressing 5T4 17–25 -specific TCRs were tested for recognition of tumor target lines in a 4-h cytotoxicity assay with 10:1 E:T. For each target cell-line, 5T4 17–25 -specific TCR transduced effector CD8 + T-cells were plotted in the following order: HD_A-2, HD_A-15, HD_B-19, HD_B-21, HD_C-3, HD_C-17, KCD_D-6, and untransduced. a Targets are 5T4 + /HLA-A2 + RCC cell lines (A498, BB65, TREP, DOBSKI; red bars), the 5T4 + /HLA-A2 − RCC line (SST548; open bars) and the 5T4 − /HLA-A2 + LCL (BB65-LCL; gray shading). b Targets are 5T4 + /HLA-A2 + breast cancer line (MDA231; blue bars), a 5T4 + /HLA-A2 − breast cancer line (BT20; open bars) and 5T4 + /HLA-A2 + colon cancer line (SW480, green bars). CD8 + T-cells with (gTRAC_KO) or without (gCtrl) native TRA disruption expressing 5T4 p17 -specific TCRs were tested for recognition of c 5T4 + /HLA-A2 + RCC cell-lines (A498, BB65, DOBSKI), the 5T4 + /HLA-A2 − RCC line (SST548), the 5T4 − /HLA-A2 + BB65-LCL, and d paired primary 5T4 + /HLA-A2 + RCC and autologous PTEC cell-lines from four RCC patients

    Journal: Cancer Immunology, Immunotherapy

    Article Title: Preclinical development of T-cell receptor-engineered T-cell therapy targeting the 5T4 tumor antigen on renal cell carcinoma

    doi: 10.1007/s00262-019-02419-4

    Figure Lengend Snippet: Cytotoxicity of 5T4 p17 -specific TCR-transduced CD8 + T-cells against 5T4-expressing tumor targets. CD8 + T-cells expressing 5T4 17–25 -specific TCRs were tested for recognition of tumor target lines in a 4-h cytotoxicity assay with 10:1 E:T. For each target cell-line, 5T4 17–25 -specific TCR transduced effector CD8 + T-cells were plotted in the following order: HD_A-2, HD_A-15, HD_B-19, HD_B-21, HD_C-3, HD_C-17, KCD_D-6, and untransduced. a Targets are 5T4 + /HLA-A2 + RCC cell lines (A498, BB65, TREP, DOBSKI; red bars), the 5T4 + /HLA-A2 − RCC line (SST548; open bars) and the 5T4 − /HLA-A2 + LCL (BB65-LCL; gray shading). b Targets are 5T4 + /HLA-A2 + breast cancer line (MDA231; blue bars), a 5T4 + /HLA-A2 − breast cancer line (BT20; open bars) and 5T4 + /HLA-A2 + colon cancer line (SW480, green bars). CD8 + T-cells with (gTRAC_KO) or without (gCtrl) native TRA disruption expressing 5T4 p17 -specific TCRs were tested for recognition of c 5T4 + /HLA-A2 + RCC cell-lines (A498, BB65, DOBSKI), the 5T4 + /HLA-A2 − RCC line (SST548), the 5T4 − /HLA-A2 + BB65-LCL, and d paired primary 5T4 + /HLA-A2 + RCC and autologous PTEC cell-lines from four RCC patients

    Article Snippet: To confirm 5T4 expression on surface after MVA-5T4 infection, target cells were stained with a 5T4-specific mAb (clone 524744; R&D systems, Minneapolis, MN) at 1 μg/mL followed by a 1:50 dilution of a secondary PE-labeled anti-mouse IgG1 mAb (clone A85-1; BD Biosciences) for analysis by flow cytometry.

    Techniques: Expressing, Cytotoxicity Assay, Disruption

    Cytotoxicity of 5T4 p17 -specific TCR-transduced CD8 + T-cells against MVA-5T4 infected targets. a Schematic of human 5T4 protein, p17–25 is in the signal sequence region. Trans: transmembrane region, Cyto: cytoplasmic region. b T2 cells and BB65-LCL were infected with MVA-WT or MVA-5T4, and cell surface expression of 5T4 was analyzed by immunostaining and flow cytometry 24 h after infection. The fraction of cells positive for cell surface 5T4 after MVA-5T4 infection is indicated. CD8 + T-cells expressing 5T4 p17 -specific TCRs were tested for recognition of c T2 and d BB65-LCL cells infected by MVA-WT, MVA-5T4, or uninfected cells pulsed with 10 nM 5T4 p17 peptide in a 6-h cytotoxicity assay at a 10:1 E:T

    Journal: Cancer Immunology, Immunotherapy

    Article Title: Preclinical development of T-cell receptor-engineered T-cell therapy targeting the 5T4 tumor antigen on renal cell carcinoma

    doi: 10.1007/s00262-019-02419-4

    Figure Lengend Snippet: Cytotoxicity of 5T4 p17 -specific TCR-transduced CD8 + T-cells against MVA-5T4 infected targets. a Schematic of human 5T4 protein, p17–25 is in the signal sequence region. Trans: transmembrane region, Cyto: cytoplasmic region. b T2 cells and BB65-LCL were infected with MVA-WT or MVA-5T4, and cell surface expression of 5T4 was analyzed by immunostaining and flow cytometry 24 h after infection. The fraction of cells positive for cell surface 5T4 after MVA-5T4 infection is indicated. CD8 + T-cells expressing 5T4 p17 -specific TCRs were tested for recognition of c T2 and d BB65-LCL cells infected by MVA-WT, MVA-5T4, or uninfected cells pulsed with 10 nM 5T4 p17 peptide in a 6-h cytotoxicity assay at a 10:1 E:T

    Article Snippet: To confirm 5T4 expression on surface after MVA-5T4 infection, target cells were stained with a 5T4-specific mAb (clone 524744; R&D systems, Minneapolis, MN) at 1 μg/mL followed by a 1:50 dilution of a secondary PE-labeled anti-mouse IgG1 mAb (clone A85-1; BD Biosciences) for analysis by flow cytometry.

    Techniques: Infection, Sequencing, Expressing, Immunostaining, Flow Cytometry, Cytotoxicity Assay