t1b 196 cd4 cd70 b cells  (ATCC)


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    ATCC t1b 196 cd4 cd70 b cells
    Schematic diagram showing conventional bivalent monospecific <t>anti-CD4</t> and <t>anti-CD70</t> IgGs along with anti-CD4/CD70 monovalent bispecific DuetMab.
    T1b 196 Cd4 Cd70 B Cells, supplied by ATCC, 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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    1) Product Images from "Insights into the molecular basis of a bispecific antibody's target selectivity"

    Article Title: Insights into the molecular basis of a bispecific antibody's target selectivity

    Journal: mAbs

    doi: 10.1080/19420862.2015.1022695

    Schematic diagram showing conventional bivalent monospecific anti-CD4 and anti-CD70 IgGs along with anti-CD4/CD70 monovalent bispecific DuetMab.
    Figure Legend Snippet: Schematic diagram showing conventional bivalent monospecific anti-CD4 and anti-CD70 IgGs along with anti-CD4/CD70 monovalent bispecific DuetMab.

    Techniques Used:

    Cell binding and ADCC activity of anti-CD4/CD70 DuetMab. ( A ) Anti-CD4/CD70 DuetMab exhibits preferential cell binding to CD4 + /CD70 + T cells via concurrent engagement to CD4 and CD70 on a single cell. ( B ) Anti-CD4/CD70 DuetMab preferentially kills CD4 + /CD70 + T cells as measured by ADCC. Each point represents the mean value of triplicate wells and the standard deviation is represented by error bars.
    Figure Legend Snippet: Cell binding and ADCC activity of anti-CD4/CD70 DuetMab. ( A ) Anti-CD4/CD70 DuetMab exhibits preferential cell binding to CD4 + /CD70 + T cells via concurrent engagement to CD4 and CD70 on a single cell. ( B ) Anti-CD4/CD70 DuetMab preferentially kills CD4 + /CD70 + T cells as measured by ADCC. Each point represents the mean value of triplicate wells and the standard deviation is represented by error bars.

    Techniques Used: Binding Assay, Activity Assay, Standard Deviation

     CD4  and  CD70  receptor density on human lymphocytes
    Figure Legend Snippet: CD4 and CD70 receptor density on human lymphocytes

    Techniques Used:

    Binding affinity of IgG and DuetMab to  CD4  and  CD70
    Figure Legend Snippet: Binding affinity of IgG and DuetMab to CD4 and CD70

    Techniques Used: Binding Assay

    Cell binding of various DuetMabs variants. Binding of anti-CD4/CD70 DuetMab variants to ( A ) CD4 + /CD70 + , ( B ) CD4 − /CD70 + and ( C ) CD4 + /CD70 − lymphocytes in a mixture of all 3 cell types. All variants with reduced affinity to CD4 exhibited improved binding selectivity over the parental DuetMab whereas their binding to target CD4 + /CD70 + T cells was not substantially impaired. Each point represents the mean values of triplicate wells and the standard deviation is represented by error bars.
    Figure Legend Snippet: Cell binding of various DuetMabs variants. Binding of anti-CD4/CD70 DuetMab variants to ( A ) CD4 + /CD70 + , ( B ) CD4 − /CD70 + and ( C ) CD4 + /CD70 − lymphocytes in a mixture of all 3 cell types. All variants with reduced affinity to CD4 exhibited improved binding selectivity over the parental DuetMab whereas their binding to target CD4 + /CD70 + T cells was not substantially impaired. Each point represents the mean values of triplicate wells and the standard deviation is represented by error bars.

    Techniques Used: Binding Assay, Standard Deviation

    ADCC activity of various DuetMabs variants. ( A ) Selective ADCC depletion of CD4 + /CD70 + T cells in a cell-mixture also containing non-target CD4 + /CD70 − T cells and CD4 − /CD70 + B cells. ( B ) ADCC activity of parental and anti-CD4 VκY94A+V H Y99A/CD70 DuetMabs against individual populations of CD4 + /CD70 + and CD4 + /CD70 − T-lymphocytes at varying E:T ratios. Each point in these studies represents the mean values of triplicate wells and the standard deviation is represented by error bars.
    Figure Legend Snippet: ADCC activity of various DuetMabs variants. ( A ) Selective ADCC depletion of CD4 + /CD70 + T cells in a cell-mixture also containing non-target CD4 + /CD70 − T cells and CD4 − /CD70 + B cells. ( B ) ADCC activity of parental and anti-CD4 VκY94A+V H Y99A/CD70 DuetMabs against individual populations of CD4 + /CD70 + and CD4 + /CD70 − T-lymphocytes at varying E:T ratios. Each point in these studies represents the mean values of triplicate wells and the standard deviation is represented by error bars.

    Techniques Used: Activity Assay, Standard Deviation

    Cytotoxicity of various  CD4  affinity-reduced DuetMab variants
    Figure Legend Snippet: Cytotoxicity of various CD4 affinity-reduced DuetMab variants

    Techniques Used:

    Effect of antibody valence on cell binding and ADCC activity. ( A ) Cell binding and ( B ) ADCC activity of anti-CD4 VκY94A+V H Y99A/CD70 and 2 monospecific (anti-CD4 VκY94A+V H Y99A/NMGC and anti-CD70/NMGC) DuetMabs at equimolar concentration against CD4 + /CD70 + T cells alone. ( C ) Non-target ADCC activity of anti-CD4 variants formatted as either monovalent anti-CD4/CD70 DuetMab or bivalent anti-CD4 IgG against CD4 + /CD70 − T cells alone. Each point in these studies represents the mean values of triplicate wells and the standard deviation is represented by error bars.
    Figure Legend Snippet: Effect of antibody valence on cell binding and ADCC activity. ( A ) Cell binding and ( B ) ADCC activity of anti-CD4 VκY94A+V H Y99A/CD70 and 2 monospecific (anti-CD4 VκY94A+V H Y99A/NMGC and anti-CD70/NMGC) DuetMabs at equimolar concentration against CD4 + /CD70 + T cells alone. ( C ) Non-target ADCC activity of anti-CD4 variants formatted as either monovalent anti-CD4/CD70 DuetMab or bivalent anti-CD4 IgG against CD4 + /CD70 − T cells alone. Each point in these studies represents the mean values of triplicate wells and the standard deviation is represented by error bars.

    Techniques Used: Binding Assay, Activity Assay, Concentration Assay, Standard Deviation

    n a cd70 transgene expressing k32  (ATCC)


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    ATCC n a cd70 transgene expressing k32
    N A Cd70 Transgene Expressing K32, supplied by ATCC, 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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    mouse full length cd70 surface antien  (ATCC)


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    ATCC mouse full length cd70 surface antien
    Mouse Full Length Cd70 Surface Antien, supplied by ATCC, 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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    cd70  (ATCC)


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    ATCC cd70
    Cd70, supplied by ATCC, 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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    cd70 expression  (ATCC)


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    ATCC cd70 expression
    In vitro characterization of [ 68 <t>Ga]Ga-NOTA-anti-CD70</t> VHH. A Analysis of CD70 protein expression measured by flow cytometry using in the upper trace anti-CD70 VHH-Hilyte Fluor 488 (107B8) or control VHH-Hilyte Fluor 488 and in the lower trace PE mouse anti-human CD70 or PE mouse IgG3 isotype control. B Relative amount of cell associated activity of [ 68 Ga]Ga-NOTA-anti-CD70 VHH (5 nM, 0.04 µg) on CD70 high and CD70 low cells (1 × 10 6 ), or in the presence of excess unlabeled VHH (50 µg, block). (C) Specific binding of [ 68 Ga]Ga-NOTA-anti-CD70 VHH on Raji cells (5 × 10 5 ). (**** p < 0.0001)
    Cd70 Expression, supplied by ATCC, 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 "Site-specific 68 Ga-labeled nanobody for PET imaging of CD70 expression in preclinical tumor models"

    Article Title: Site-specific 68 Ga-labeled nanobody for PET imaging of CD70 expression in preclinical tumor models

    Journal: EJNMMI Radiopharmacy and Chemistry

    doi: 10.1186/s41181-023-00194-3

    In vitro characterization of [ 68 Ga]Ga-NOTA-anti-CD70 VHH. A Analysis of CD70 protein expression measured by flow cytometry using in the upper trace anti-CD70 VHH-Hilyte Fluor 488 (107B8) or control VHH-Hilyte Fluor 488 and in the lower trace PE mouse anti-human CD70 or PE mouse IgG3 isotype control. B Relative amount of cell associated activity of [ 68 Ga]Ga-NOTA-anti-CD70 VHH (5 nM, 0.04 µg) on CD70 high and CD70 low cells (1 × 10 6 ), or in the presence of excess unlabeled VHH (50 µg, block). (C) Specific binding of [ 68 Ga]Ga-NOTA-anti-CD70 VHH on Raji cells (5 × 10 5 ). (**** p < 0.0001)
    Figure Legend Snippet: In vitro characterization of [ 68 Ga]Ga-NOTA-anti-CD70 VHH. A Analysis of CD70 protein expression measured by flow cytometry using in the upper trace anti-CD70 VHH-Hilyte Fluor 488 (107B8) or control VHH-Hilyte Fluor 488 and in the lower trace PE mouse anti-human CD70 or PE mouse IgG3 isotype control. B Relative amount of cell associated activity of [ 68 Ga]Ga-NOTA-anti-CD70 VHH (5 nM, 0.04 µg) on CD70 high and CD70 low cells (1 × 10 6 ), or in the presence of excess unlabeled VHH (50 µg, block). (C) Specific binding of [ 68 Ga]Ga-NOTA-anti-CD70 VHH on Raji cells (5 × 10 5 ). (**** p < 0.0001)

    Techniques Used: In Vitro, Expressing, Flow Cytometry, Activity Assay, Blocking Assay, Binding Assay

    Biodistribution analysis of [ 68 Ga]Ga-NOTA-anti-CD70 VHH at 15, 30, 60 and 90 min post radiotracer injection. (%ID/g = % injected dose/ gram, injected radiotracer dose = 1–4.7 µg; ~ 1.5–2.5 MBq/mouse, n = 3/time point)
    Figure Legend Snippet: Biodistribution analysis of [ 68 Ga]Ga-NOTA-anti-CD70 VHH at 15, 30, 60 and 90 min post radiotracer injection. (%ID/g = % injected dose/ gram, injected radiotracer dose = 1–4.7 µg; ~ 1.5–2.5 MBq/mouse, n = 3/time point)

    Techniques Used: Injection

    PET/CT imaging using [ 68 Ga]Ga-NOTA-anti-CD70 VHH fragment— A Representative coronal µPET/CT images (30–60 min post injection timeframe summed activity, slice thickness 0.78 mm) of mice bearing CD70 high (786-O (left panel) and with block (middle panel)) and CD70 low tumors (NCl-H1975, right panel). B Time-activity curves for radiotracer distribution in the tumors are shown. (%ID/mL = % injected dose/ mL, injected radiotracer dose = 4–6.7 µg; ~ 6.4–7.4 MBq/mouse; n = 7, data are presented as mean ± SEM. **** p < 0.0001; #### p = 0.0029)
    Figure Legend Snippet: PET/CT imaging using [ 68 Ga]Ga-NOTA-anti-CD70 VHH fragment— A Representative coronal µPET/CT images (30–60 min post injection timeframe summed activity, slice thickness 0.78 mm) of mice bearing CD70 high (786-O (left panel) and with block (middle panel)) and CD70 low tumors (NCl-H1975, right panel). B Time-activity curves for radiotracer distribution in the tumors are shown. (%ID/mL = % injected dose/ mL, injected radiotracer dose = 4–6.7 µg; ~ 6.4–7.4 MBq/mouse; n = 7, data are presented as mean ± SEM. **** p < 0.0001; #### p = 0.0029)

    Techniques Used: Positron Emission Tomography-Computed Tomography, Imaging, Injection, Activity Assay, Blocking Assay

    Ex vivo analysis of 786-O and NCl-H1975 tumors. Representative autoradiography (ARG) and microscopy images of adjacent histologic tumor slices stained for CD70 using a biotinylated anti-CD70 VHH
    Figure Legend Snippet: Ex vivo analysis of 786-O and NCl-H1975 tumors. Representative autoradiography (ARG) and microscopy images of adjacent histologic tumor slices stained for CD70 using a biotinylated anti-CD70 VHH

    Techniques Used: Ex Vivo, Autoradiography, Microscopy, Staining

    Binding affinity kinetics (equilibrium dissociation constant (K D ), association (k a ) and dissociation (k d ) rates) of  anti-CD70  VHHs as determined by SPR
    Figure Legend Snippet: Binding affinity kinetics (equilibrium dissociation constant (K D ), association (k a ) and dissociation (k d ) rates) of anti-CD70 VHHs as determined by SPR

    Techniques Used: Binding Assay

    mouse full length cd70 surface antigen  (ATCC)


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    ATCC mouse full length cd70 surface antigen
    <t>CD70</t> expression correlates with survival in patients with solid tumors. A) (top) CD70 gene expression for different solid tumors compared with normal matched tissues. Sources: The University of Alabama at Birmingham Cancer data analysis (UALCAN) and The Cancer Genome Atlas program (TCGA). (Bottom panels) survival rates of patients diagnosed with renal carcinoma and lung cancer with low and high CD70 expression. Source: The Human Protein Atlas program. B-C) Confirmation of CD70 (target) expression on cancer cells – B) glioblastoma (GBM) and C) osteosarcoma (OS) models - and (bottom) transduction of chimeric antigen receptor (CAR) construct in B) C57BL/6 mice - derived T cells (CAR T Kr158B ) and C) in Balb/c mice - derived T cells (CAR T K7M2 ) as indicated by GFP reporter. On average, D) percentage of CD70 expression is 73% for glioblastoma and 99% for osteosarcoma models. E) Mean transduction of 66 % for CAR T Kr158B and 60% for CAR T K7M2 . The name of the cell lines and tumor models will be used interchangeably – GBM for Kr158B and OS for K7M2.
    Mouse Full Length Cd70 Surface Antigen, supplied by ATCC, 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 "Three-Dimensional Bioconjugated Liquid-Like Solid (LLS) Enhance Characterization of Solid Tumor - Chimeric Antigen Receptor T cell interactions"

    Article Title: Three-Dimensional Bioconjugated Liquid-Like Solid (LLS) Enhance Characterization of Solid Tumor - Chimeric Antigen Receptor T cell interactions

    Journal: bioRxiv

    doi: 10.1101/2023.02.17.529033

    CD70 expression correlates with survival in patients with solid tumors. A) (top) CD70 gene expression for different solid tumors compared with normal matched tissues. Sources: The University of Alabama at Birmingham Cancer data analysis (UALCAN) and The Cancer Genome Atlas program (TCGA). (Bottom panels) survival rates of patients diagnosed with renal carcinoma and lung cancer with low and high CD70 expression. Source: The Human Protein Atlas program. B-C) Confirmation of CD70 (target) expression on cancer cells – B) glioblastoma (GBM) and C) osteosarcoma (OS) models - and (bottom) transduction of chimeric antigen receptor (CAR) construct in B) C57BL/6 mice - derived T cells (CAR T Kr158B ) and C) in Balb/c mice - derived T cells (CAR T K7M2 ) as indicated by GFP reporter. On average, D) percentage of CD70 expression is 73% for glioblastoma and 99% for osteosarcoma models. E) Mean transduction of 66 % for CAR T Kr158B and 60% for CAR T K7M2 . The name of the cell lines and tumor models will be used interchangeably – GBM for Kr158B and OS for K7M2.
    Figure Legend Snippet: CD70 expression correlates with survival in patients with solid tumors. A) (top) CD70 gene expression for different solid tumors compared with normal matched tissues. Sources: The University of Alabama at Birmingham Cancer data analysis (UALCAN) and The Cancer Genome Atlas program (TCGA). (Bottom panels) survival rates of patients diagnosed with renal carcinoma and lung cancer with low and high CD70 expression. Source: The Human Protein Atlas program. B-C) Confirmation of CD70 (target) expression on cancer cells – B) glioblastoma (GBM) and C) osteosarcoma (OS) models - and (bottom) transduction of chimeric antigen receptor (CAR) construct in B) C57BL/6 mice - derived T cells (CAR T Kr158B ) and C) in Balb/c mice - derived T cells (CAR T K7M2 ) as indicated by GFP reporter. On average, D) percentage of CD70 expression is 73% for glioblastoma and 99% for osteosarcoma models. E) Mean transduction of 66 % for CAR T Kr158B and 60% for CAR T K7M2 . The name of the cell lines and tumor models will be used interchangeably – GBM for Kr158B and OS for K7M2.

    Techniques Used: Expressing, Transduction, Construct, Derivative Assay

    CAR T cell directed motion towards the target tumor. A) Representative confocal snapshots of CAR T cells infiltrating a GBM solid tumor in 3D. The panel shows CD70-specific CAR T cells (green) navigating through the supported COL1-LLS RhB microgels (red) and infiltrating the target GBM tumor (white). Blue arrows indicate paths of CAR T migration. B, D) mean velocity of CAR T cells cocultured with CD70 pos tumors and the WT control for B) GBM and D) OS tumor models. The number of tracks (n) is indicated on the plots. The number of biological replicates is N=2. An unpaired two-tailed Student’s t-test was performed. Statistical significance with p values was indicated on the plots. C, E) Quantifying tumor-infiltrating CAR T cells on average from 0-72h as a percent of total CAR T cells for C) GBM and E) OS tumors. The box plots display 25 th and 75 th percentiles, a line at the median, and a plus sign at the mean, from the minimum to the maximum observation. Unpaired two-tailed Student’s t-test was performed (n=234, N=3, statistical significance with p values were indicated on the plots). F) Top panel: Maximum intensity z projection (top panel) showing snapshots of CAR T – GBM tumor interaction at 0, 24, and 72h. Middle panel: the segmentation of CAR T with colors indicated by individual CAR T cells at each frame. The segmentation employed a deep learning-based method (as discussed in the method section). Bottom panel: maximum intensity projection of the segmented CAR T cell velocity tracks overtime. The velocity gradient was color-coded, showing accumulation of CAR T inside the tumor. The segmented cells were tracked using Linear Assignment Problem (LAP) tracker at the maximum frame-to-frame linking and allowable track segment gap closing of 150 μm (~ 3 cell diameter). G) Evidence of chemotaxis and upregulation in migratory pathways for CAR T cells co-cultured with their target tumors for GBM (top panel) and OS (bottom panel). Noticeably, evidence of immune-mediated cytotoxic function is demonstrated via IFNγ detection.
    Figure Legend Snippet: CAR T cell directed motion towards the target tumor. A) Representative confocal snapshots of CAR T cells infiltrating a GBM solid tumor in 3D. The panel shows CD70-specific CAR T cells (green) navigating through the supported COL1-LLS RhB microgels (red) and infiltrating the target GBM tumor (white). Blue arrows indicate paths of CAR T migration. B, D) mean velocity of CAR T cells cocultured with CD70 pos tumors and the WT control for B) GBM and D) OS tumor models. The number of tracks (n) is indicated on the plots. The number of biological replicates is N=2. An unpaired two-tailed Student’s t-test was performed. Statistical significance with p values was indicated on the plots. C, E) Quantifying tumor-infiltrating CAR T cells on average from 0-72h as a percent of total CAR T cells for C) GBM and E) OS tumors. The box plots display 25 th and 75 th percentiles, a line at the median, and a plus sign at the mean, from the minimum to the maximum observation. Unpaired two-tailed Student’s t-test was performed (n=234, N=3, statistical significance with p values were indicated on the plots). F) Top panel: Maximum intensity z projection (top panel) showing snapshots of CAR T – GBM tumor interaction at 0, 24, and 72h. Middle panel: the segmentation of CAR T with colors indicated by individual CAR T cells at each frame. The segmentation employed a deep learning-based method (as discussed in the method section). Bottom panel: maximum intensity projection of the segmented CAR T cell velocity tracks overtime. The velocity gradient was color-coded, showing accumulation of CAR T inside the tumor. The segmented cells were tracked using Linear Assignment Problem (LAP) tracker at the maximum frame-to-frame linking and allowable track segment gap closing of 150 μm (~ 3 cell diameter). G) Evidence of chemotaxis and upregulation in migratory pathways for CAR T cells co-cultured with their target tumors for GBM (top panel) and OS (bottom panel). Noticeably, evidence of immune-mediated cytotoxic function is demonstrated via IFNγ detection.

    Techniques Used: Migration, Two Tailed Test, Chemotaxis Assay, Cell Culture

    CAR T expansion, activation, and killing. A) Representative confocal timelapse images of the FITC channel (green) that show immune activation, expansion, and killing of the target tumors. The patterns of CAR T clustering and rapid expansion were observed in almost all conditions with efficient anti-tumor activity. White arrows locate CAR T cell clusters. B, C) The number of CAR T clusters rapidly increased during the first 24h and steadily maintained for more than 72h in B) GBM and C) OS models. D, E) Cluster size in all cocultures with target tumors steadily increased over time but not in the WT controls. F, G) Expansion of CAR T clusters revealed an inverse correlation with tumor size. (n=3 for all CD70 pos samples and n=2 for all WT samples). H, I) Endpoint flow cytometry data measuring CAR T expansion after 96h of CAR T – cancer cells coculture in the 2D assay for both tumor models. J, K) A comparison in IFNγ secretion (pg/mL) for 2D vs 3D for J) GBM and K) OS. The number of biological replicates is N=2. L) Top row: confocal 3D snapshots of GBM tumors after co-cultured with CAR T cells for 24h showing highly tortuous tumor margin of CD70 pos tumor as compared to the WT counterpart. Bottom row: cross-section view (z depth: 70 μm) of the sample on the top row exposing infiltrating CAR T cells within the tumor mass. M) Measurement of tumor tortuosity factors revealed more than 3-fold change for the CD70 pos tumors within the first 48h of coculture. The tumor tortuosity factor was calculated as a ratio between the perimeter of the tumor outline to the perimeter of a circle of the same pixel area. Data was obtained from GBM samples (n=3) at an initial E:T ratio of 1:2, and from osteosarcoma (K7M2) samples (n=3) at an initial E:T ratio of 1:1. Biological samples for each group (n= 3), and technical repetitions (n= 3) were performed.
    Figure Legend Snippet: CAR T expansion, activation, and killing. A) Representative confocal timelapse images of the FITC channel (green) that show immune activation, expansion, and killing of the target tumors. The patterns of CAR T clustering and rapid expansion were observed in almost all conditions with efficient anti-tumor activity. White arrows locate CAR T cell clusters. B, C) The number of CAR T clusters rapidly increased during the first 24h and steadily maintained for more than 72h in B) GBM and C) OS models. D, E) Cluster size in all cocultures with target tumors steadily increased over time but not in the WT controls. F, G) Expansion of CAR T clusters revealed an inverse correlation with tumor size. (n=3 for all CD70 pos samples and n=2 for all WT samples). H, I) Endpoint flow cytometry data measuring CAR T expansion after 96h of CAR T – cancer cells coculture in the 2D assay for both tumor models. J, K) A comparison in IFNγ secretion (pg/mL) for 2D vs 3D for J) GBM and K) OS. The number of biological replicates is N=2. L) Top row: confocal 3D snapshots of GBM tumors after co-cultured with CAR T cells for 24h showing highly tortuous tumor margin of CD70 pos tumor as compared to the WT counterpart. Bottom row: cross-section view (z depth: 70 μm) of the sample on the top row exposing infiltrating CAR T cells within the tumor mass. M) Measurement of tumor tortuosity factors revealed more than 3-fold change for the CD70 pos tumors within the first 48h of coculture. The tumor tortuosity factor was calculated as a ratio between the perimeter of the tumor outline to the perimeter of a circle of the same pixel area. Data was obtained from GBM samples (n=3) at an initial E:T ratio of 1:2, and from osteosarcoma (K7M2) samples (n=3) at an initial E:T ratio of 1:1. Biological samples for each group (n= 3), and technical repetitions (n= 3) were performed.

    Techniques Used: Activation Assay, Activity Assay, Flow Cytometry, Two-Dimensional Assay, Cell Culture

    Sensitivity of anti-tumor activity to various CAR T: cancer cell (E:T) ratios. A,B) confocal time-lapse images of CAR T – tumor co-culture in iVITA at different E:T ratios. C,D) show the digital image reconstruction of confocal data quantifying bulk tumor mass, migrating single cancer cells, immune cells, and immune cell clusters, and killing activity over time. A,C) GBM CD70 pos tumors and B,D) OS CD70 pos tumors were co-cultured with their respective CAR T cells at different concentrations corresponding to E:T of 1:4, 1:2, and 4:1 for the GBM model and E:T of 1:4, 1:1, and 4:1 for the OS model. At 72h and E:T= 4:1, almost 100% tumor elimination was observed in both models. E-H) CAR T expansion as a function of initial E:T seeding ratios. E,F) CAR T expansion on average in both models from 0-72h. For each seeding E:T, CAR T expansion at each time point was normalized to the CAR T at time 0, and the average was calculated for all frames. G,H) the number of CAR T clusters counted every 1.5h for 72h for each group. The box plots display 25th and 75th percentiles, a line at the median a plus sign at the mean, from the minimum to the maximum observation. I,J) Quantification of tumor size over time at the initial E:T= 1:4, 1:2, and 4:1 for I) GBM model and at E:T= 1:4, 1:1, and 4:1 for J) OS model. K,L) The E:T ratio dynamically changed over time. The CAR T expansion and tumor-killing were presented by the exponential increase of E:T ratios. M,N) tumor killing rates were calculated as derivatives from I) and J), respectively. Statistical analysis was performed using Ordinary One-Way ANOVA. (n=3 unless indicated otherwise, ** = p < 0.01, and **** = p < 0.0001).
    Figure Legend Snippet: Sensitivity of anti-tumor activity to various CAR T: cancer cell (E:T) ratios. A,B) confocal time-lapse images of CAR T – tumor co-culture in iVITA at different E:T ratios. C,D) show the digital image reconstruction of confocal data quantifying bulk tumor mass, migrating single cancer cells, immune cells, and immune cell clusters, and killing activity over time. A,C) GBM CD70 pos tumors and B,D) OS CD70 pos tumors were co-cultured with their respective CAR T cells at different concentrations corresponding to E:T of 1:4, 1:2, and 4:1 for the GBM model and E:T of 1:4, 1:1, and 4:1 for the OS model. At 72h and E:T= 4:1, almost 100% tumor elimination was observed in both models. E-H) CAR T expansion as a function of initial E:T seeding ratios. E,F) CAR T expansion on average in both models from 0-72h. For each seeding E:T, CAR T expansion at each time point was normalized to the CAR T at time 0, and the average was calculated for all frames. G,H) the number of CAR T clusters counted every 1.5h for 72h for each group. The box plots display 25th and 75th percentiles, a line at the median a plus sign at the mean, from the minimum to the maximum observation. I,J) Quantification of tumor size over time at the initial E:T= 1:4, 1:2, and 4:1 for I) GBM model and at E:T= 1:4, 1:1, and 4:1 for J) OS model. K,L) The E:T ratio dynamically changed over time. The CAR T expansion and tumor-killing were presented by the exponential increase of E:T ratios. M,N) tumor killing rates were calculated as derivatives from I) and J), respectively. Statistical analysis was performed using Ordinary One-Way ANOVA. (n=3 unless indicated otherwise, ** = p < 0.01, and **** = p < 0.0001).

    Techniques Used: Activity Assay, Co-Culture Assay, Cell Culture

    tite tri specific molecule cd19 cd3 cd70 tsm m  (ATCC)


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    ATCC tite tri specific molecule cd19 cd3 cd70 tsm m
    Tite Tri Specific Molecule Cd19 Cd3 Cd70 Tsm M, supplied by ATCC, 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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    tite tri specific molecule cd19 cd3 cd70 tsm d  (ATCC)


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    ATCC tite tri specific molecule cd19 cd3 cd70 tsm d
    Tite Tri Specific Molecule Cd19 Cd3 Cd70 Tsm D, supplied by ATCC, 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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    cd70  (ATCC)


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    ATCC cd70
    (A) Two plasmids – pMAZ-IgH and pMAZ-IgL – were used as backbones to construct and express <t>anti-CD70</t> immunocytokines bearing either murine (m) or human (h) IFN-γ. pMAZ-IgH expresses the anti-CD70 heavy chain separated from murine or human IFN-γ by a flexible (Gly) 4 -Ser linker. pMAZ-IgL encodes the anti-CD70 light chain. For details of construction, expression and purification, please see the Materials and Methods section. (B) Coomassie Blue-stained SDS-PAGE gel of mIFN-γ-anti-CD70 immunocytokine (lane 1), and hIFN-γ-anti-CD70 immunocytokine (lane 2) purified from supernatants of 293T cells after transfection with the plasmids described in A.
    Cd70, supplied by ATCC, 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 "Anti-CD70 Immunocytokines for Exploitation of Interferon-γ-Induced RIP1-Dependent Necrosis in Renal Cell Carcinoma"

    Article Title: Anti-CD70 Immunocytokines for Exploitation of Interferon-γ-Induced RIP1-Dependent Necrosis in Renal Cell Carcinoma

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0061446

    (A) Two plasmids – pMAZ-IgH and pMAZ-IgL – were used as backbones to construct and express anti-CD70 immunocytokines bearing either murine (m) or human (h) IFN-γ. pMAZ-IgH expresses the anti-CD70 heavy chain separated from murine or human IFN-γ by a flexible (Gly) 4 -Ser linker. pMAZ-IgL encodes the anti-CD70 light chain. For details of construction, expression and purification, please see the Materials and Methods section. (B) Coomassie Blue-stained SDS-PAGE gel of mIFN-γ-anti-CD70 immunocytokine (lane 1), and hIFN-γ-anti-CD70 immunocytokine (lane 2) purified from supernatants of 293T cells after transfection with the plasmids described in A.
    Figure Legend Snippet: (A) Two plasmids – pMAZ-IgH and pMAZ-IgL – were used as backbones to construct and express anti-CD70 immunocytokines bearing either murine (m) or human (h) IFN-γ. pMAZ-IgH expresses the anti-CD70 heavy chain separated from murine or human IFN-γ by a flexible (Gly) 4 -Ser linker. pMAZ-IgL encodes the anti-CD70 light chain. For details of construction, expression and purification, please see the Materials and Methods section. (B) Coomassie Blue-stained SDS-PAGE gel of mIFN-γ-anti-CD70 immunocytokine (lane 1), and hIFN-γ-anti-CD70 immunocytokine (lane 2) purified from supernatants of 293T cells after transfection with the plasmids described in A.

    Techniques Used: Construct, Expressing, Purification, Staining, SDS Page, Transfection

    Murine (RenCa) or human (Caki-1) RCC cells were treated with anti-CD70 immunocytokines bearing either murine (m) or human (h) IFN-γ (‘Anti-CD70 fusion’, 50 ng/ml). As controls, parallel populations of these cells were treated with recombinant murine or human IFN-γ (‘Native’, 10 ng/ml). At the indicated times post-treatment, cells were examined by immunoblotting for either phosphorylated (p-STAT1) or total STAT1.
    Figure Legend Snippet: Murine (RenCa) or human (Caki-1) RCC cells were treated with anti-CD70 immunocytokines bearing either murine (m) or human (h) IFN-γ (‘Anti-CD70 fusion’, 50 ng/ml). As controls, parallel populations of these cells were treated with recombinant murine or human IFN-γ (‘Native’, 10 ng/ml). At the indicated times post-treatment, cells were examined by immunoblotting for either phosphorylated (p-STAT1) or total STAT1.

    Techniques Used: Recombinant, Western Blot

    Murine (RenCa) or human (Caki-1) RCC cells were pre-treated for 16 h with anti-CD70 immunocytokines bearing either murine (m) or human (h) IFN-γ (‘Anti-CD70 fusion’, 50 ng/ml). As controls, parallel populations of these cells were pre-treated for 16 h with recombinant murine or human IFN-γ (‘Native’, 10 ng/ml), or with unfused anti-CD70 antibody (50 ng/ml). Following pre-treatment, cells were infected with VSV-GFP (MOI = 5 for RenCa, 0.05 for Caki-1). (A) Infected cells were photographed by brightfield (for demonstration of cytopathic effect) or by fluorescence (to show viral replication) microscopy 20 h post-infection. (B) Viability of cells treated as above was determined 20 h post-infection. (C) VSV progeny yield from supernatants of infected cells was determined by standard plaque assay 20 h post-infection. Error bars represent mean +/− S.D, n = 3.
    Figure Legend Snippet: Murine (RenCa) or human (Caki-1) RCC cells were pre-treated for 16 h with anti-CD70 immunocytokines bearing either murine (m) or human (h) IFN-γ (‘Anti-CD70 fusion’, 50 ng/ml). As controls, parallel populations of these cells were pre-treated for 16 h with recombinant murine or human IFN-γ (‘Native’, 10 ng/ml), or with unfused anti-CD70 antibody (50 ng/ml). Following pre-treatment, cells were infected with VSV-GFP (MOI = 5 for RenCa, 0.05 for Caki-1). (A) Infected cells were photographed by brightfield (for demonstration of cytopathic effect) or by fluorescence (to show viral replication) microscopy 20 h post-infection. (B) Viability of cells treated as above was determined 20 h post-infection. (C) VSV progeny yield from supernatants of infected cells was determined by standard plaque assay 20 h post-infection. Error bars represent mean +/− S.D, n = 3.

    Techniques Used: Recombinant, Infection, Fluorescence, Microscopy, Plaque Assay

    (A) 293T cells were transfected with an expression vector encoding Myc-tagged human CD70 (‘CD70’), or with an empty vector (‘Vec’). 24 h post-transfection, cells were examined for CD70 expression in lysates by anti-Myc immunoblotting (inset, top panel; β-actin loading control, bottom panel), or on the cell surface by FACS staining with a FITC-conjugated anti-CD70 monoclonal antibody. (B) 293T cells were transfected as in A with either an empty vector (‘Vec’) or an expression vector encoding Myc-tagged CD70 (‘CD70’). 24 h post-transfection, cells were incubated with either Rituximab as an isotype control human IgG1 antibody (‘Isotype Control’, left panel), or with immunocytokines bearing either murine (m) or human (h) IFN-γ (‘Anti-CD70 fusion’), and, following labeling with FITC-conjugated anti-human IgG secondary antibodies, analyzed by FACS for CD70 expression. (C) The ATCC-derived RCC cell lines 786-O, 769-P, Caki-1, and ACHN were incubated with either an isotype control human IgG1 antibody (Rituximab, dashed line), anti-CD70-mIFN-γ immunocytokine (thin solid line), or anti-CD70-hIFN-γ immunocytokine (thick solid line), followed by labeling with FITC-conjugated anti-human IgG secondary antibodies and detection of fluorescence by FACS. All four ATCC cell lines are robustly and specifically stained by both anti-CD70 IFN-γ immunocytokines.
    Figure Legend Snippet: (A) 293T cells were transfected with an expression vector encoding Myc-tagged human CD70 (‘CD70’), or with an empty vector (‘Vec’). 24 h post-transfection, cells were examined for CD70 expression in lysates by anti-Myc immunoblotting (inset, top panel; β-actin loading control, bottom panel), or on the cell surface by FACS staining with a FITC-conjugated anti-CD70 monoclonal antibody. (B) 293T cells were transfected as in A with either an empty vector (‘Vec’) or an expression vector encoding Myc-tagged CD70 (‘CD70’). 24 h post-transfection, cells were incubated with either Rituximab as an isotype control human IgG1 antibody (‘Isotype Control’, left panel), or with immunocytokines bearing either murine (m) or human (h) IFN-γ (‘Anti-CD70 fusion’), and, following labeling with FITC-conjugated anti-human IgG secondary antibodies, analyzed by FACS for CD70 expression. (C) The ATCC-derived RCC cell lines 786-O, 769-P, Caki-1, and ACHN were incubated with either an isotype control human IgG1 antibody (Rituximab, dashed line), anti-CD70-mIFN-γ immunocytokine (thin solid line), or anti-CD70-hIFN-γ immunocytokine (thick solid line), followed by labeling with FITC-conjugated anti-human IgG secondary antibodies and detection of fluorescence by FACS. All four ATCC cell lines are robustly and specifically stained by both anti-CD70 IFN-γ immunocytokines.

    Techniques Used: Transfection, Expressing, Plasmid Preparation, Western Blot, Staining, Incubation, Labeling, Derivative Assay, Fluorescence

    RCC cell lines RenCa (A) or Caki-1 (B) were treated either with unfused anti-CD70 antibody (‘Anti-CD70’), with recombinant, native human or murine IFN-γ (‘Native’), or with human or murine IFN-γ immunocytokines (‘Anti-CD70 fusion’) for 72 h in the presence of their MTD of bortezomib (black bars). As controls, these cells were also treated with each agent singly (grey bars). In conditions requiring bortezomib co-treatment, bortezomib was added to cells 1 h before IFN-γ.
    Figure Legend Snippet: RCC cell lines RenCa (A) or Caki-1 (B) were treated either with unfused anti-CD70 antibody (‘Anti-CD70’), with recombinant, native human or murine IFN-γ (‘Native’), or with human or murine IFN-γ immunocytokines (‘Anti-CD70 fusion’) for 72 h in the presence of their MTD of bortezomib (black bars). As controls, these cells were also treated with each agent singly (grey bars). In conditions requiring bortezomib co-treatment, bortezomib was added to cells 1 h before IFN-γ.

    Techniques Used: Recombinant

    (A) RenCa, Caki-1, 786-O, or HRC63 cells were co-treated with bortezomib (MTD) and, respectively, murine (RenCa) or human (Caki-1, 786-O, and HRC63) IFN-γ immunocytokines (‘Anti-CD70 fusion’, 50 ng/ml) in the presence or absence of 50 μM RIP1 kinase inhibitor Nec-1 for 72–84 h. The MTD of bortezomib for 786-0 and HRC63 cells was 4 ng/ml and 2 ng/ml, respectively. Cell viability was determined by Trypan Blue exclusion analysis. Error bars represent mean +/− S.D; n = 3. (B) RenCa, Caki-1, 786-O, or HRC63 cells pre-treated without (-Nec-1) or with (+Nec-1) for 1h, before co-treatment with IFN-γ immunocytokines and bortezomib as in (A), were photographed 72 h post-treatment.
    Figure Legend Snippet: (A) RenCa, Caki-1, 786-O, or HRC63 cells were co-treated with bortezomib (MTD) and, respectively, murine (RenCa) or human (Caki-1, 786-O, and HRC63) IFN-γ immunocytokines (‘Anti-CD70 fusion’, 50 ng/ml) in the presence or absence of 50 μM RIP1 kinase inhibitor Nec-1 for 72–84 h. The MTD of bortezomib for 786-0 and HRC63 cells was 4 ng/ml and 2 ng/ml, respectively. Cell viability was determined by Trypan Blue exclusion analysis. Error bars represent mean +/− S.D; n = 3. (B) RenCa, Caki-1, 786-O, or HRC63 cells pre-treated without (-Nec-1) or with (+Nec-1) for 1h, before co-treatment with IFN-γ immunocytokines and bortezomib as in (A), were photographed 72 h post-treatment.

    Techniques Used:

    cd70  (ATCC)


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    ATCC cd70
    (A) Two plasmids – pMAZ-IgH and pMAZ-IgL – were used as backbones to construct and express <t>anti-CD70</t> immunocytokines bearing either murine (m) or human (h) IFN-γ. pMAZ-IgH expresses the anti-CD70 heavy chain separated from murine or human IFN-γ by a flexible (Gly) 4 -Ser linker. pMAZ-IgL encodes the anti-CD70 light chain. For details of construction, expression and purification, please see the Materials and Methods section. (B) Coomassie Blue-stained SDS-PAGE gel of mIFN-γ-anti-CD70 immunocytokine (lane 1), and hIFN-γ-anti-CD70 immunocytokine (lane 2) purified from supernatants of 293T cells after transfection with the plasmids described in A.
    Cd70, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cd70/product/ATCC
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    cd70 - by Bioz Stars, 2024-04
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    Images

    1) Product Images from "Anti-CD70 Immunocytokines for Exploitation of Interferon-γ-Induced RIP1-Dependent Necrosis in Renal Cell Carcinoma"

    Article Title: Anti-CD70 Immunocytokines for Exploitation of Interferon-γ-Induced RIP1-Dependent Necrosis in Renal Cell Carcinoma

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0061446

    (A) Two plasmids – pMAZ-IgH and pMAZ-IgL – were used as backbones to construct and express anti-CD70 immunocytokines bearing either murine (m) or human (h) IFN-γ. pMAZ-IgH expresses the anti-CD70 heavy chain separated from murine or human IFN-γ by a flexible (Gly) 4 -Ser linker. pMAZ-IgL encodes the anti-CD70 light chain. For details of construction, expression and purification, please see the Materials and Methods section. (B) Coomassie Blue-stained SDS-PAGE gel of mIFN-γ-anti-CD70 immunocytokine (lane 1), and hIFN-γ-anti-CD70 immunocytokine (lane 2) purified from supernatants of 293T cells after transfection with the plasmids described in A.
    Figure Legend Snippet: (A) Two plasmids – pMAZ-IgH and pMAZ-IgL – were used as backbones to construct and express anti-CD70 immunocytokines bearing either murine (m) or human (h) IFN-γ. pMAZ-IgH expresses the anti-CD70 heavy chain separated from murine or human IFN-γ by a flexible (Gly) 4 -Ser linker. pMAZ-IgL encodes the anti-CD70 light chain. For details of construction, expression and purification, please see the Materials and Methods section. (B) Coomassie Blue-stained SDS-PAGE gel of mIFN-γ-anti-CD70 immunocytokine (lane 1), and hIFN-γ-anti-CD70 immunocytokine (lane 2) purified from supernatants of 293T cells after transfection with the plasmids described in A.

    Techniques Used: Construct, Expressing, Purification, Staining, SDS Page, Transfection

    Murine (RenCa) or human (Caki-1) RCC cells were treated with anti-CD70 immunocytokines bearing either murine (m) or human (h) IFN-γ (‘Anti-CD70 fusion’, 50 ng/ml). As controls, parallel populations of these cells were treated with recombinant murine or human IFN-γ (‘Native’, 10 ng/ml). At the indicated times post-treatment, cells were examined by immunoblotting for either phosphorylated (p-STAT1) or total STAT1.
    Figure Legend Snippet: Murine (RenCa) or human (Caki-1) RCC cells were treated with anti-CD70 immunocytokines bearing either murine (m) or human (h) IFN-γ (‘Anti-CD70 fusion’, 50 ng/ml). As controls, parallel populations of these cells were treated with recombinant murine or human IFN-γ (‘Native’, 10 ng/ml). At the indicated times post-treatment, cells were examined by immunoblotting for either phosphorylated (p-STAT1) or total STAT1.

    Techniques Used: Recombinant, Western Blot

    Murine (RenCa) or human (Caki-1) RCC cells were pre-treated for 16 h with anti-CD70 immunocytokines bearing either murine (m) or human (h) IFN-γ (‘Anti-CD70 fusion’, 50 ng/ml). As controls, parallel populations of these cells were pre-treated for 16 h with recombinant murine or human IFN-γ (‘Native’, 10 ng/ml), or with unfused anti-CD70 antibody (50 ng/ml). Following pre-treatment, cells were infected with VSV-GFP (MOI = 5 for RenCa, 0.05 for Caki-1). (A) Infected cells were photographed by brightfield (for demonstration of cytopathic effect) or by fluorescence (to show viral replication) microscopy 20 h post-infection. (B) Viability of cells treated as above was determined 20 h post-infection. (C) VSV progeny yield from supernatants of infected cells was determined by standard plaque assay 20 h post-infection. Error bars represent mean +/− S.D, n = 3.
    Figure Legend Snippet: Murine (RenCa) or human (Caki-1) RCC cells were pre-treated for 16 h with anti-CD70 immunocytokines bearing either murine (m) or human (h) IFN-γ (‘Anti-CD70 fusion’, 50 ng/ml). As controls, parallel populations of these cells were pre-treated for 16 h with recombinant murine or human IFN-γ (‘Native’, 10 ng/ml), or with unfused anti-CD70 antibody (50 ng/ml). Following pre-treatment, cells were infected with VSV-GFP (MOI = 5 for RenCa, 0.05 for Caki-1). (A) Infected cells were photographed by brightfield (for demonstration of cytopathic effect) or by fluorescence (to show viral replication) microscopy 20 h post-infection. (B) Viability of cells treated as above was determined 20 h post-infection. (C) VSV progeny yield from supernatants of infected cells was determined by standard plaque assay 20 h post-infection. Error bars represent mean +/− S.D, n = 3.

    Techniques Used: Recombinant, Infection, Fluorescence, Microscopy, Plaque Assay

    (A) 293T cells were transfected with an expression vector encoding Myc-tagged human CD70 (‘CD70’), or with an empty vector (‘Vec’). 24 h post-transfection, cells were examined for CD70 expression in lysates by anti-Myc immunoblotting (inset, top panel; β-actin loading control, bottom panel), or on the cell surface by FACS staining with a FITC-conjugated anti-CD70 monoclonal antibody. (B) 293T cells were transfected as in A with either an empty vector (‘Vec’) or an expression vector encoding Myc-tagged CD70 (‘CD70’). 24 h post-transfection, cells were incubated with either Rituximab as an isotype control human IgG1 antibody (‘Isotype Control’, left panel), or with immunocytokines bearing either murine (m) or human (h) IFN-γ (‘Anti-CD70 fusion’), and, following labeling with FITC-conjugated anti-human IgG secondary antibodies, analyzed by FACS for CD70 expression. (C) The ATCC-derived RCC cell lines 786-O, 769-P, Caki-1, and ACHN were incubated with either an isotype control human IgG1 antibody (Rituximab, dashed line), anti-CD70-mIFN-γ immunocytokine (thin solid line), or anti-CD70-hIFN-γ immunocytokine (thick solid line), followed by labeling with FITC-conjugated anti-human IgG secondary antibodies and detection of fluorescence by FACS. All four ATCC cell lines are robustly and specifically stained by both anti-CD70 IFN-γ immunocytokines.
    Figure Legend Snippet: (A) 293T cells were transfected with an expression vector encoding Myc-tagged human CD70 (‘CD70’), or with an empty vector (‘Vec’). 24 h post-transfection, cells were examined for CD70 expression in lysates by anti-Myc immunoblotting (inset, top panel; β-actin loading control, bottom panel), or on the cell surface by FACS staining with a FITC-conjugated anti-CD70 monoclonal antibody. (B) 293T cells were transfected as in A with either an empty vector (‘Vec’) or an expression vector encoding Myc-tagged CD70 (‘CD70’). 24 h post-transfection, cells were incubated with either Rituximab as an isotype control human IgG1 antibody (‘Isotype Control’, left panel), or with immunocytokines bearing either murine (m) or human (h) IFN-γ (‘Anti-CD70 fusion’), and, following labeling with FITC-conjugated anti-human IgG secondary antibodies, analyzed by FACS for CD70 expression. (C) The ATCC-derived RCC cell lines 786-O, 769-P, Caki-1, and ACHN were incubated with either an isotype control human IgG1 antibody (Rituximab, dashed line), anti-CD70-mIFN-γ immunocytokine (thin solid line), or anti-CD70-hIFN-γ immunocytokine (thick solid line), followed by labeling with FITC-conjugated anti-human IgG secondary antibodies and detection of fluorescence by FACS. All four ATCC cell lines are robustly and specifically stained by both anti-CD70 IFN-γ immunocytokines.

    Techniques Used: Transfection, Expressing, Plasmid Preparation, Western Blot, Staining, Incubation, Labeling, Derivative Assay, Fluorescence

    RCC cell lines RenCa (A) or Caki-1 (B) were treated either with unfused anti-CD70 antibody (‘Anti-CD70’), with recombinant, native human or murine IFN-γ (‘Native’), or with human or murine IFN-γ immunocytokines (‘Anti-CD70 fusion’) for 72 h in the presence of their MTD of bortezomib (black bars). As controls, these cells were also treated with each agent singly (grey bars). In conditions requiring bortezomib co-treatment, bortezomib was added to cells 1 h before IFN-γ.
    Figure Legend Snippet: RCC cell lines RenCa (A) or Caki-1 (B) were treated either with unfused anti-CD70 antibody (‘Anti-CD70’), with recombinant, native human or murine IFN-γ (‘Native’), or with human or murine IFN-γ immunocytokines (‘Anti-CD70 fusion’) for 72 h in the presence of their MTD of bortezomib (black bars). As controls, these cells were also treated with each agent singly (grey bars). In conditions requiring bortezomib co-treatment, bortezomib was added to cells 1 h before IFN-γ.

    Techniques Used: Recombinant

    (A) RenCa, Caki-1, 786-O, or HRC63 cells were co-treated with bortezomib (MTD) and, respectively, murine (RenCa) or human (Caki-1, 786-O, and HRC63) IFN-γ immunocytokines (‘Anti-CD70 fusion’, 50 ng/ml) in the presence or absence of 50 μM RIP1 kinase inhibitor Nec-1 for 72–84 h. The MTD of bortezomib for 786-0 and HRC63 cells was 4 ng/ml and 2 ng/ml, respectively. Cell viability was determined by Trypan Blue exclusion analysis. Error bars represent mean +/− S.D; n = 3. (B) RenCa, Caki-1, 786-O, or HRC63 cells pre-treated without (-Nec-1) or with (+Nec-1) for 1h, before co-treatment with IFN-γ immunocytokines and bortezomib as in (A), were photographed 72 h post-treatment.
    Figure Legend Snippet: (A) RenCa, Caki-1, 786-O, or HRC63 cells were co-treated with bortezomib (MTD) and, respectively, murine (RenCa) or human (Caki-1, 786-O, and HRC63) IFN-γ immunocytokines (‘Anti-CD70 fusion’, 50 ng/ml) in the presence or absence of 50 μM RIP1 kinase inhibitor Nec-1 for 72–84 h. The MTD of bortezomib for 786-0 and HRC63 cells was 4 ng/ml and 2 ng/ml, respectively. Cell viability was determined by Trypan Blue exclusion analysis. Error bars represent mean +/− S.D; n = 3. (B) RenCa, Caki-1, 786-O, or HRC63 cells pre-treated without (-Nec-1) or with (+Nec-1) for 1h, before co-treatment with IFN-γ immunocytokines and bortezomib as in (A), were photographed 72 h post-treatment.

    Techniques Used:

    t1b 196 cd4 cd70 b cells  (ATCC)


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    ATCC t1b 196 cd4 cd70 b cells
    Schematic diagram showing conventional bivalent monospecific <t>anti-CD4</t> and <t>anti-CD70</t> IgGs along with anti-CD4/CD70 monovalent bispecific DuetMab.
    T1b 196 Cd4 Cd70 B Cells, supplied by ATCC, 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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    1) Product Images from "Insights into the molecular basis of a bispecific antibody's target selectivity"

    Article Title: Insights into the molecular basis of a bispecific antibody's target selectivity

    Journal: mAbs

    doi: 10.1080/19420862.2015.1022695

    Schematic diagram showing conventional bivalent monospecific anti-CD4 and anti-CD70 IgGs along with anti-CD4/CD70 monovalent bispecific DuetMab.
    Figure Legend Snippet: Schematic diagram showing conventional bivalent monospecific anti-CD4 and anti-CD70 IgGs along with anti-CD4/CD70 monovalent bispecific DuetMab.

    Techniques Used:

    Cell binding and ADCC activity of anti-CD4/CD70 DuetMab. ( A ) Anti-CD4/CD70 DuetMab exhibits preferential cell binding to CD4 + /CD70 + T cells via concurrent engagement to CD4 and CD70 on a single cell. ( B ) Anti-CD4/CD70 DuetMab preferentially kills CD4 + /CD70 + T cells as measured by ADCC. Each point represents the mean value of triplicate wells and the standard deviation is represented by error bars.
    Figure Legend Snippet: Cell binding and ADCC activity of anti-CD4/CD70 DuetMab. ( A ) Anti-CD4/CD70 DuetMab exhibits preferential cell binding to CD4 + /CD70 + T cells via concurrent engagement to CD4 and CD70 on a single cell. ( B ) Anti-CD4/CD70 DuetMab preferentially kills CD4 + /CD70 + T cells as measured by ADCC. Each point represents the mean value of triplicate wells and the standard deviation is represented by error bars.

    Techniques Used: Binding Assay, Activity Assay, Standard Deviation

     CD4  and  CD70  receptor density on human lymphocytes
    Figure Legend Snippet: CD4 and CD70 receptor density on human lymphocytes

    Techniques Used:

    Binding affinity of IgG and DuetMab to  CD4  and  CD70
    Figure Legend Snippet: Binding affinity of IgG and DuetMab to CD4 and CD70

    Techniques Used: Binding Assay

    Cell binding of various DuetMabs variants. Binding of anti-CD4/CD70 DuetMab variants to ( A ) CD4 + /CD70 + , ( B ) CD4 − /CD70 + and ( C ) CD4 + /CD70 − lymphocytes in a mixture of all 3 cell types. All variants with reduced affinity to CD4 exhibited improved binding selectivity over the parental DuetMab whereas their binding to target CD4 + /CD70 + T cells was not substantially impaired. Each point represents the mean values of triplicate wells and the standard deviation is represented by error bars.
    Figure Legend Snippet: Cell binding of various DuetMabs variants. Binding of anti-CD4/CD70 DuetMab variants to ( A ) CD4 + /CD70 + , ( B ) CD4 − /CD70 + and ( C ) CD4 + /CD70 − lymphocytes in a mixture of all 3 cell types. All variants with reduced affinity to CD4 exhibited improved binding selectivity over the parental DuetMab whereas their binding to target CD4 + /CD70 + T cells was not substantially impaired. Each point represents the mean values of triplicate wells and the standard deviation is represented by error bars.

    Techniques Used: Binding Assay, Standard Deviation

    ADCC activity of various DuetMabs variants. ( A ) Selective ADCC depletion of CD4 + /CD70 + T cells in a cell-mixture also containing non-target CD4 + /CD70 − T cells and CD4 − /CD70 + B cells. ( B ) ADCC activity of parental and anti-CD4 VκY94A+V H Y99A/CD70 DuetMabs against individual populations of CD4 + /CD70 + and CD4 + /CD70 − T-lymphocytes at varying E:T ratios. Each point in these studies represents the mean values of triplicate wells and the standard deviation is represented by error bars.
    Figure Legend Snippet: ADCC activity of various DuetMabs variants. ( A ) Selective ADCC depletion of CD4 + /CD70 + T cells in a cell-mixture also containing non-target CD4 + /CD70 − T cells and CD4 − /CD70 + B cells. ( B ) ADCC activity of parental and anti-CD4 VκY94A+V H Y99A/CD70 DuetMabs against individual populations of CD4 + /CD70 + and CD4 + /CD70 − T-lymphocytes at varying E:T ratios. Each point in these studies represents the mean values of triplicate wells and the standard deviation is represented by error bars.

    Techniques Used: Activity Assay, Standard Deviation

    Cytotoxicity of various  CD4  affinity-reduced DuetMab variants
    Figure Legend Snippet: Cytotoxicity of various CD4 affinity-reduced DuetMab variants

    Techniques Used:

    Effect of antibody valence on cell binding and ADCC activity. ( A ) Cell binding and ( B ) ADCC activity of anti-CD4 VκY94A+V H Y99A/CD70 and 2 monospecific (anti-CD4 VκY94A+V H Y99A/NMGC and anti-CD70/NMGC) DuetMabs at equimolar concentration against CD4 + /CD70 + T cells alone. ( C ) Non-target ADCC activity of anti-CD4 variants formatted as either monovalent anti-CD4/CD70 DuetMab or bivalent anti-CD4 IgG against CD4 + /CD70 − T cells alone. Each point in these studies represents the mean values of triplicate wells and the standard deviation is represented by error bars.
    Figure Legend Snippet: Effect of antibody valence on cell binding and ADCC activity. ( A ) Cell binding and ( B ) ADCC activity of anti-CD4 VκY94A+V H Y99A/CD70 and 2 monospecific (anti-CD4 VκY94A+V H Y99A/NMGC and anti-CD70/NMGC) DuetMabs at equimolar concentration against CD4 + /CD70 + T cells alone. ( C ) Non-target ADCC activity of anti-CD4 variants formatted as either monovalent anti-CD4/CD70 DuetMab or bivalent anti-CD4 IgG against CD4 + /CD70 − T cells alone. Each point in these studies represents the mean values of triplicate wells and the standard deviation is represented by error bars.

    Techniques Used: Binding Assay, Activity Assay, Concentration Assay, Standard Deviation

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