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human her2 positive her2þ breast cancer cell line skbr3  (ATCC)


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    ATCC human her2 positive her2þ breast cancer cell line skbr3
    Figure 1. TNF regulation of CD137 expression in NK cells upon ADCC. A, Relative expression of TNFRSF9, TNFRSF18,TNFRSF4,TNFRSF1B,TNFRSF8,CTLA4,and PDCD1 bygene expression microarrays of resting (NT) and CD16-activated NK cells (12 hours after activation with anti-CD16). Data from 2 donors (D1, D2). B–G, NK cells were cocultured with <t>SKBR3</t> (SK) and trastuzumab (Tz; 210 ng/mL) in the presence or absence of the anti-TNFa infliximab (Inflix; 50 mg/mL), recombinant human TNFa (rTNFa; 10 ng/mL), anti–TNF-RI (a-RI; 10 mg/mL), anti–TNF-RII (a-RII; 10 mg/mL), or control mouse IgG1 antibody (cIgG1; 10 mg/mL) for 24 hours. B, Mean SEM proportions of CD137þ, OX40þ, and PD1þ NK cells at 24 hours after ADCC, as analyzed by flow cytometry. Data from three independent experiments. C, Representative dot plots of CD137, OX40, PD1, and CD16 expression by NK cells before and after ADCC. Inset numbers indicate proportions of markerþ NK cells. Data from a representative experiment out of three performed. D, Mean SEM of surface CD137 levels on NK cells upon ADCC assays in the presence of infliximab or recombinant human TNFa. Data normalized to trastuzumab-activated NK cells. Data from four independent experiments. E, Kinetics of CD137þ NK cells upon trastuzumab-induced ADCC in the presence or absence of infliximab. Graph represents mean SEM data from four to six experiments. h, hours. F, Surface TNF-RI and TNF-RII staining on resting (NK; solid line) and trastuzumab-activated NK cells (NKþSKBR3þTz; dashed line). Control staining (Iso ctrl; filled histogram). G, Mean SEM surface CD137 levels on NK cells upon ADCC in the presence or absence of TNF/TNFR blocking antibodies. Data normalized to trastuzumab-activated NK cells in the absence of blocking antibodies (-). Data from four independent experiments. Statistical significance was calculated using paired or unpaired Student t test for nonnormalized or normalized data, respectively (, P < 0.05; , P < 0.01; , P < 0.001; , P < 0.0001). MFI, mean fluorescence intensity.
    Human Her2 Positive Her2þ Breast Cancer Cell Line Skbr3, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 6550 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "CD137 Costimulation Counteracts TGFβ Inhibition of NK-cell Antitumor Function"

    Article Title: CD137 Costimulation Counteracts TGFβ Inhibition of NK-cell Antitumor Function

    Journal: Cancer Immunology Research

    doi: 10.1158/2326-6066.cir-21-0030

    Figure 1. TNF regulation of CD137 expression in NK cells upon ADCC. A, Relative expression of TNFRSF9, TNFRSF18,TNFRSF4,TNFRSF1B,TNFRSF8,CTLA4,and PDCD1 bygene expression microarrays of resting (NT) and CD16-activated NK cells (12 hours after activation with anti-CD16). Data from 2 donors (D1, D2). B–G, NK cells were cocultured with SKBR3 (SK) and trastuzumab (Tz; 210 ng/mL) in the presence or absence of the anti-TNFa infliximab (Inflix; 50 mg/mL), recombinant human TNFa (rTNFa; 10 ng/mL), anti–TNF-RI (a-RI; 10 mg/mL), anti–TNF-RII (a-RII; 10 mg/mL), or control mouse IgG1 antibody (cIgG1; 10 mg/mL) for 24 hours. B, Mean SEM proportions of CD137þ, OX40þ, and PD1þ NK cells at 24 hours after ADCC, as analyzed by flow cytometry. Data from three independent experiments. C, Representative dot plots of CD137, OX40, PD1, and CD16 expression by NK cells before and after ADCC. Inset numbers indicate proportions of markerþ NK cells. Data from a representative experiment out of three performed. D, Mean SEM of surface CD137 levels on NK cells upon ADCC assays in the presence of infliximab or recombinant human TNFa. Data normalized to trastuzumab-activated NK cells. Data from four independent experiments. E, Kinetics of CD137þ NK cells upon trastuzumab-induced ADCC in the presence or absence of infliximab. Graph represents mean SEM data from four to six experiments. h, hours. F, Surface TNF-RI and TNF-RII staining on resting (NK; solid line) and trastuzumab-activated NK cells (NKþSKBR3þTz; dashed line). Control staining (Iso ctrl; filled histogram). G, Mean SEM surface CD137 levels on NK cells upon ADCC in the presence or absence of TNF/TNFR blocking antibodies. Data normalized to trastuzumab-activated NK cells in the absence of blocking antibodies (-). Data from four independent experiments. Statistical significance was calculated using paired or unpaired Student t test for nonnormalized or normalized data, respectively (, P < 0.05; , P < 0.01; , P < 0.001; , P < 0.0001). MFI, mean fluorescence intensity.
    Figure Legend Snippet: Figure 1. TNF regulation of CD137 expression in NK cells upon ADCC. A, Relative expression of TNFRSF9, TNFRSF18,TNFRSF4,TNFRSF1B,TNFRSF8,CTLA4,and PDCD1 bygene expression microarrays of resting (NT) and CD16-activated NK cells (12 hours after activation with anti-CD16). Data from 2 donors (D1, D2). B–G, NK cells were cocultured with SKBR3 (SK) and trastuzumab (Tz; 210 ng/mL) in the presence or absence of the anti-TNFa infliximab (Inflix; 50 mg/mL), recombinant human TNFa (rTNFa; 10 ng/mL), anti–TNF-RI (a-RI; 10 mg/mL), anti–TNF-RII (a-RII; 10 mg/mL), or control mouse IgG1 antibody (cIgG1; 10 mg/mL) for 24 hours. B, Mean SEM proportions of CD137þ, OX40þ, and PD1þ NK cells at 24 hours after ADCC, as analyzed by flow cytometry. Data from three independent experiments. C, Representative dot plots of CD137, OX40, PD1, and CD16 expression by NK cells before and after ADCC. Inset numbers indicate proportions of markerþ NK cells. Data from a representative experiment out of three performed. D, Mean SEM of surface CD137 levels on NK cells upon ADCC assays in the presence of infliximab or recombinant human TNFa. Data normalized to trastuzumab-activated NK cells. Data from four independent experiments. E, Kinetics of CD137þ NK cells upon trastuzumab-induced ADCC in the presence or absence of infliximab. Graph represents mean SEM data from four to six experiments. h, hours. F, Surface TNF-RI and TNF-RII staining on resting (NK; solid line) and trastuzumab-activated NK cells (NKþSKBR3þTz; dashed line). Control staining (Iso ctrl; filled histogram). G, Mean SEM surface CD137 levels on NK cells upon ADCC in the presence or absence of TNF/TNFR blocking antibodies. Data normalized to trastuzumab-activated NK cells in the absence of blocking antibodies (-). Data from four independent experiments. Statistical significance was calculated using paired or unpaired Student t test for nonnormalized or normalized data, respectively (, P < 0.05; , P < 0.01; , P < 0.001; , P < 0.0001). MFI, mean fluorescence intensity.

    Techniques Used: Expressing, Activation Assay, Recombinant, Control, Cytometry, Staining, Blocking Assay

    Figure 4. Impact of CD137 costimulation in the function of TGFb-treated NK cells. NK cells activated via CD16 in the presence/absence of TGFb1 with or without urelumab (Ure; 50 or 1 mg/mL) or control human IgG4 (cIgG4; 50 mg/mL) for 6 days were used as effector cells in restimulation experiments. A, Dot plots showing surface CD107a on NK cells rechallenged with K562 for 4 hours. Data from a representative experiment out of three performed. Inset numbers indicate the percentage of CD107aþ NK cells. B, Mean SEM of CD107aþ NK-cell frequencies upon rechallenge with K562 for 4 hours in three independent experiments. C–E, NK cells were rechallenged with SKBR3 cellswith and without trastuzumab for 4 hours.C, Dot plots showing activated caspase 3 (aCasp3) in SKBR3 cells after 4-hour ADCC with restimulated NK cells and trastuzumab. Inset numbers indicate the percentage of active caspase 3þ SKBR3 cells. (Continued on the following page.)
    Figure Legend Snippet: Figure 4. Impact of CD137 costimulation in the function of TGFb-treated NK cells. NK cells activated via CD16 in the presence/absence of TGFb1 with or without urelumab (Ure; 50 or 1 mg/mL) or control human IgG4 (cIgG4; 50 mg/mL) for 6 days were used as effector cells in restimulation experiments. A, Dot plots showing surface CD107a on NK cells rechallenged with K562 for 4 hours. Data from a representative experiment out of three performed. Inset numbers indicate the percentage of CD107aþ NK cells. B, Mean SEM of CD107aþ NK-cell frequencies upon rechallenge with K562 for 4 hours in three independent experiments. C–E, NK cells were rechallenged with SKBR3 cellswith and without trastuzumab for 4 hours.C, Dot plots showing activated caspase 3 (aCasp3) in SKBR3 cells after 4-hour ADCC with restimulated NK cells and trastuzumab. Inset numbers indicate the percentage of active caspase 3þ SKBR3 cells. (Continued on the following page.)

    Techniques Used: Control

    Figure 7. NK-cell signature, NK-cell effector molecules, CD137 axis, and TGFb expression in treatment-naive breast carcinomas and their association with response to trastuzumab. Levels of the gene signature predicting NK-cell infiltration (see “Materials and Methods”), IFNG, GZMB, CCL5, FCGR3A (CD16), KLRK1 (NKG2D), CXCL9, CXCL10, TGFB1–3, as well as TNFRSF9 (CD137) and TNFSF9 (CD137L) were analyzed in treatment-na€ve HER2þ breast carcinomas from a phase II trial dataset (GSE130786). A, Violin plot showing NK-cell GSVA scores in tumors from patients achieving pCR to trastuzumab neoadjuvant treatment (n ¼ 40) or not (n ¼ 41). TI, tumor-infiltrating. B, Correlation between tumor-infiltrating NK-cell score and GZMB, CCL5, IFNG, and TGFB1–3 scores. Statistical significance calculated by Pearson test. C, Correlation between tumor-infiltrating NK-cell score and TNFRSF9. D, Correlation of TNFRSF9 and TNFSF9 between themselves and with NK-cell effector molecules. Statistical significance calculated by Pearson test. E, Association of CCL5, GZMB, IFNG, TNFRSF9, TNFSF9, KLRK1, FCGR3A, CXCL10, and CXCL9 scores in tumors from patients achieving pCR to trastuzumab or not. Statistical significance by unpaired Student t test. F, OR and 97.5% CI of NK cell, IFNG, GZMB, CCL5, TGFB1, TGFB2, and TGFB3 scores for pCR to trastuzumab.
    Figure Legend Snippet: Figure 7. NK-cell signature, NK-cell effector molecules, CD137 axis, and TGFb expression in treatment-naive breast carcinomas and their association with response to trastuzumab. Levels of the gene signature predicting NK-cell infiltration (see “Materials and Methods”), IFNG, GZMB, CCL5, FCGR3A (CD16), KLRK1 (NKG2D), CXCL9, CXCL10, TGFB1–3, as well as TNFRSF9 (CD137) and TNFSF9 (CD137L) were analyzed in treatment-na€ve HER2þ breast carcinomas from a phase II trial dataset (GSE130786). A, Violin plot showing NK-cell GSVA scores in tumors from patients achieving pCR to trastuzumab neoadjuvant treatment (n ¼ 40) or not (n ¼ 41). TI, tumor-infiltrating. B, Correlation between tumor-infiltrating NK-cell score and GZMB, CCL5, IFNG, and TGFB1–3 scores. Statistical significance calculated by Pearson test. C, Correlation between tumor-infiltrating NK-cell score and TNFRSF9. D, Correlation of TNFRSF9 and TNFSF9 between themselves and with NK-cell effector molecules. Statistical significance calculated by Pearson test. E, Association of CCL5, GZMB, IFNG, TNFRSF9, TNFSF9, KLRK1, FCGR3A, CXCL10, and CXCL9 scores in tumors from patients achieving pCR to trastuzumab or not. Statistical significance by unpaired Student t test. F, OR and 97.5% CI of NK cell, IFNG, GZMB, CCL5, TGFB1, TGFB2, and TGFB3 scores for pCR to trastuzumab.

    Techniques Used: Expressing

    Related Articles

    Expressing:

    Article Title: CD137 Costimulation Counteracts TGFβ Inhibition of NK-cell Antitumor Function
    Article Snippet: The human HER2-positive (HER2þ) breast cancer cell line SKBR3 (ATCC catalog no. HTB-30, RRID:CVCL_0033) was obtained from ATCC.. SKBR3 cells were grown in DMEM/F-12 medium (SigmaAldrich, catalog no. D6421) supplemented with L-glutamine (0.5 mmol/L, Gibco, catalog no. 25030–024).SKBR3 cells were grown in DMEM/F-12 medium (SigmaAldrich, catalog no. D6421) supplemented with L-glutamine (0.5 mmol/L, Gibco, catalog no. 25030–024).

    Activation Assay:

    Article Title: CD137 Costimulation Counteracts TGFβ Inhibition of NK-cell Antitumor Function
    Article Snippet: The human HER2-positive (HER2þ) breast cancer cell line SKBR3 (ATCC catalog no. HTB-30, RRID:CVCL_0033) was obtained from ATCC.. SKBR3 cells were grown in DMEM/F-12 medium (SigmaAldrich, catalog no. D6421) supplemented with L-glutamine (0.5 mmol/L, Gibco, catalog no. 25030–024).SKBR3 cells were grown in DMEM/F-12 medium (SigmaAldrich, catalog no. D6421) supplemented with L-glutamine (0.5 mmol/L, Gibco, catalog no. 25030–024).

    Recombinant:

    Article Title: CD137 Costimulation Counteracts TGFβ Inhibition of NK-cell Antitumor Function
    Article Snippet: The human HER2-positive (HER2þ) breast cancer cell line SKBR3 (ATCC catalog no. HTB-30, RRID:CVCL_0033) was obtained from ATCC.. SKBR3 cells were grown in DMEM/F-12 medium (SigmaAldrich, catalog no. D6421) supplemented with L-glutamine (0.5 mmol/L, Gibco, catalog no. 25030–024).SKBR3 cells were grown in DMEM/F-12 medium (SigmaAldrich, catalog no. D6421) supplemented with L-glutamine (0.5 mmol/L, Gibco, catalog no. 25030–024).

    Control:

    Article Title: CD137 Costimulation Counteracts TGFβ Inhibition of NK-cell Antitumor Function
    Article Snippet: The human HER2-positive (HER2þ) breast cancer cell line SKBR3 (ATCC catalog no. HTB-30, RRID:CVCL_0033) was obtained from ATCC.. SKBR3 cells were grown in DMEM/F-12 medium (SigmaAldrich, catalog no. D6421) supplemented with L-glutamine (0.5 mmol/L, Gibco, catalog no. 25030–024).SKBR3 cells were grown in DMEM/F-12 medium (SigmaAldrich, catalog no. D6421) supplemented with L-glutamine (0.5 mmol/L, Gibco, catalog no. 25030–024).

    Cytometry:

    Article Title: CD137 Costimulation Counteracts TGFβ Inhibition of NK-cell Antitumor Function
    Article Snippet: The human HER2-positive (HER2þ) breast cancer cell line SKBR3 (ATCC catalog no. HTB-30, RRID:CVCL_0033) was obtained from ATCC.. SKBR3 cells were grown in DMEM/F-12 medium (SigmaAldrich, catalog no. D6421) supplemented with L-glutamine (0.5 mmol/L, Gibco, catalog no. 25030–024).SKBR3 cells were grown in DMEM/F-12 medium (SigmaAldrich, catalog no. D6421) supplemented with L-glutamine (0.5 mmol/L, Gibco, catalog no. 25030–024).

    Staining:

    Article Title: CD137 Costimulation Counteracts TGFβ Inhibition of NK-cell Antitumor Function
    Article Snippet: The human HER2-positive (HER2þ) breast cancer cell line SKBR3 (ATCC catalog no. HTB-30, RRID:CVCL_0033) was obtained from ATCC.. SKBR3 cells were grown in DMEM/F-12 medium (SigmaAldrich, catalog no. D6421) supplemented with L-glutamine (0.5 mmol/L, Gibco, catalog no. 25030–024).SKBR3 cells were grown in DMEM/F-12 medium (SigmaAldrich, catalog no. D6421) supplemented with L-glutamine (0.5 mmol/L, Gibco, catalog no. 25030–024).

    Blocking Assay:

    Article Title: CD137 Costimulation Counteracts TGFβ Inhibition of NK-cell Antitumor Function
    Article Snippet: The human HER2-positive (HER2þ) breast cancer cell line SKBR3 (ATCC catalog no. HTB-30, RRID:CVCL_0033) was obtained from ATCC.. SKBR3 cells were grown in DMEM/F-12 medium (SigmaAldrich, catalog no. D6421) supplemented with L-glutamine (0.5 mmol/L, Gibco, catalog no. 25030–024).SKBR3 cells were grown in DMEM/F-12 medium (SigmaAldrich, catalog no. D6421) supplemented with L-glutamine (0.5 mmol/L, Gibco, catalog no. 25030–024).



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    human her2 positive her2þ breast cancer cell line skbr3  (ATCC)
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    ATCC human her2 positive her2þ breast cancer cell line skbr3
    Figure 1. TNF regulation of CD137 expression in NK cells upon ADCC. A, Relative expression of TNFRSF9, TNFRSF18,TNFRSF4,TNFRSF1B,TNFRSF8,CTLA4,and PDCD1 bygene expression microarrays of resting (NT) and CD16-activated NK cells (12 hours after activation with anti-CD16). Data from 2 donors (D1, D2). B–G, NK cells were cocultured with <t>SKBR3</t> (SK) and trastuzumab (Tz; 210 ng/mL) in the presence or absence of the anti-TNFa infliximab (Inflix; 50 mg/mL), recombinant human TNFa (rTNFa; 10 ng/mL), anti–TNF-RI (a-RI; 10 mg/mL), anti–TNF-RII (a-RII; 10 mg/mL), or control mouse IgG1 antibody (cIgG1; 10 mg/mL) for 24 hours. B, Mean SEM proportions of CD137þ, OX40þ, and PD1þ NK cells at 24 hours after ADCC, as analyzed by flow cytometry. Data from three independent experiments. C, Representative dot plots of CD137, OX40, PD1, and CD16 expression by NK cells before and after ADCC. Inset numbers indicate proportions of markerþ NK cells. Data from a representative experiment out of three performed. D, Mean SEM of surface CD137 levels on NK cells upon ADCC assays in the presence of infliximab or recombinant human TNFa. Data normalized to trastuzumab-activated NK cells. Data from four independent experiments. E, Kinetics of CD137þ NK cells upon trastuzumab-induced ADCC in the presence or absence of infliximab. Graph represents mean SEM data from four to six experiments. h, hours. F, Surface TNF-RI and TNF-RII staining on resting (NK; solid line) and trastuzumab-activated NK cells (NKþSKBR3þTz; dashed line). Control staining (Iso ctrl; filled histogram). G, Mean SEM surface CD137 levels on NK cells upon ADCC in the presence or absence of TNF/TNFR blocking antibodies. Data normalized to trastuzumab-activated NK cells in the absence of blocking antibodies (-). Data from four independent experiments. Statistical significance was calculated using paired or unpaired Student t test for nonnormalized or normalized data, respectively (, P < 0.05; , P < 0.01; , P < 0.001; , P < 0.0001). MFI, mean fluorescence intensity.
    Human Her2 Positive Her2þ Breast Cancer Cell Line Skbr3, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/human+her2+positive+her2%C3%BE+breast+cancer+cell+line+skbr3/SK-BR-3/10__1158_slash_2326___6066__cir___21___0030-44-1-9
    Average 99 stars, based on 1 article reviews
    human her2 positive her2þ breast cancer cell line skbr3 - by Bioz Stars, 2026-11
    99/100 stars
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    Figure 1. TNF regulation of CD137 expression in NK cells upon ADCC. A, Relative expression of TNFRSF9, TNFRSF18,TNFRSF4,TNFRSF1B,TNFRSF8,CTLA4,and PDCD1 bygene expression microarrays of resting (NT) and CD16-activated NK cells (12 hours after activation with anti-CD16). Data from 2 donors (D1, D2). B–G, NK cells were cocultured with SKBR3 (SK) and trastuzumab (Tz; 210 ng/mL) in the presence or absence of the anti-TNFa infliximab (Inflix; 50 mg/mL), recombinant human TNFa (rTNFa; 10 ng/mL), anti–TNF-RI (a-RI; 10 mg/mL), anti–TNF-RII (a-RII; 10 mg/mL), or control mouse IgG1 antibody (cIgG1; 10 mg/mL) for 24 hours. B, Mean SEM proportions of CD137þ, OX40þ, and PD1þ NK cells at 24 hours after ADCC, as analyzed by flow cytometry. Data from three independent experiments. C, Representative dot plots of CD137, OX40, PD1, and CD16 expression by NK cells before and after ADCC. Inset numbers indicate proportions of markerþ NK cells. Data from a representative experiment out of three performed. D, Mean SEM of surface CD137 levels on NK cells upon ADCC assays in the presence of infliximab or recombinant human TNFa. Data normalized to trastuzumab-activated NK cells. Data from four independent experiments. E, Kinetics of CD137þ NK cells upon trastuzumab-induced ADCC in the presence or absence of infliximab. Graph represents mean SEM data from four to six experiments. h, hours. F, Surface TNF-RI and TNF-RII staining on resting (NK; solid line) and trastuzumab-activated NK cells (NKþSKBR3þTz; dashed line). Control staining (Iso ctrl; filled histogram). G, Mean SEM surface CD137 levels on NK cells upon ADCC in the presence or absence of TNF/TNFR blocking antibodies. Data normalized to trastuzumab-activated NK cells in the absence of blocking antibodies (-). Data from four independent experiments. Statistical significance was calculated using paired or unpaired Student t test for nonnormalized or normalized data, respectively (, P < 0.05; , P < 0.01; , P < 0.001; , P < 0.0001). MFI, mean fluorescence intensity.

    Journal: Cancer Immunology Research

    Article Title: CD137 Costimulation Counteracts TGFβ Inhibition of NK-cell Antitumor Function

    doi: 10.1158/2326-6066.cir-21-0030

    Figure Lengend Snippet: Figure 1. TNF regulation of CD137 expression in NK cells upon ADCC. A, Relative expression of TNFRSF9, TNFRSF18,TNFRSF4,TNFRSF1B,TNFRSF8,CTLA4,and PDCD1 bygene expression microarrays of resting (NT) and CD16-activated NK cells (12 hours after activation with anti-CD16). Data from 2 donors (D1, D2). B–G, NK cells were cocultured with SKBR3 (SK) and trastuzumab (Tz; 210 ng/mL) in the presence or absence of the anti-TNFa infliximab (Inflix; 50 mg/mL), recombinant human TNFa (rTNFa; 10 ng/mL), anti–TNF-RI (a-RI; 10 mg/mL), anti–TNF-RII (a-RII; 10 mg/mL), or control mouse IgG1 antibody (cIgG1; 10 mg/mL) for 24 hours. B, Mean SEM proportions of CD137þ, OX40þ, and PD1þ NK cells at 24 hours after ADCC, as analyzed by flow cytometry. Data from three independent experiments. C, Representative dot plots of CD137, OX40, PD1, and CD16 expression by NK cells before and after ADCC. Inset numbers indicate proportions of markerþ NK cells. Data from a representative experiment out of three performed. D, Mean SEM of surface CD137 levels on NK cells upon ADCC assays in the presence of infliximab or recombinant human TNFa. Data normalized to trastuzumab-activated NK cells. Data from four independent experiments. E, Kinetics of CD137þ NK cells upon trastuzumab-induced ADCC in the presence or absence of infliximab. Graph represents mean SEM data from four to six experiments. h, hours. F, Surface TNF-RI and TNF-RII staining on resting (NK; solid line) and trastuzumab-activated NK cells (NKþSKBR3þTz; dashed line). Control staining (Iso ctrl; filled histogram). G, Mean SEM surface CD137 levels on NK cells upon ADCC in the presence or absence of TNF/TNFR blocking antibodies. Data normalized to trastuzumab-activated NK cells in the absence of blocking antibodies (-). Data from four independent experiments. Statistical significance was calculated using paired or unpaired Student t test for nonnormalized or normalized data, respectively (, P < 0.05; , P < 0.01; , P < 0.001; , P < 0.0001). MFI, mean fluorescence intensity.

    Article Snippet: The human HER2-positive (HER2þ) breast cancer cell line SKBR3 (ATCC catalog no. HTB-30, RRID:CVCL_0033) was obtained from ATCC.

    Techniques: Expressing, Activation Assay, Recombinant, Control, Cytometry, Staining, Blocking Assay

    Figure 4. Impact of CD137 costimulation in the function of TGFb-treated NK cells. NK cells activated via CD16 in the presence/absence of TGFb1 with or without urelumab (Ure; 50 or 1 mg/mL) or control human IgG4 (cIgG4; 50 mg/mL) for 6 days were used as effector cells in restimulation experiments. A, Dot plots showing surface CD107a on NK cells rechallenged with K562 for 4 hours. Data from a representative experiment out of three performed. Inset numbers indicate the percentage of CD107aþ NK cells. B, Mean SEM of CD107aþ NK-cell frequencies upon rechallenge with K562 for 4 hours in three independent experiments. C–E, NK cells were rechallenged with SKBR3 cellswith and without trastuzumab for 4 hours.C, Dot plots showing activated caspase 3 (aCasp3) in SKBR3 cells after 4-hour ADCC with restimulated NK cells and trastuzumab. Inset numbers indicate the percentage of active caspase 3þ SKBR3 cells. (Continued on the following page.)

    Journal: Cancer Immunology Research

    Article Title: CD137 Costimulation Counteracts TGFβ Inhibition of NK-cell Antitumor Function

    doi: 10.1158/2326-6066.cir-21-0030

    Figure Lengend Snippet: Figure 4. Impact of CD137 costimulation in the function of TGFb-treated NK cells. NK cells activated via CD16 in the presence/absence of TGFb1 with or without urelumab (Ure; 50 or 1 mg/mL) or control human IgG4 (cIgG4; 50 mg/mL) for 6 days were used as effector cells in restimulation experiments. A, Dot plots showing surface CD107a on NK cells rechallenged with K562 for 4 hours. Data from a representative experiment out of three performed. Inset numbers indicate the percentage of CD107aþ NK cells. B, Mean SEM of CD107aþ NK-cell frequencies upon rechallenge with K562 for 4 hours in three independent experiments. C–E, NK cells were rechallenged with SKBR3 cellswith and without trastuzumab for 4 hours.C, Dot plots showing activated caspase 3 (aCasp3) in SKBR3 cells after 4-hour ADCC with restimulated NK cells and trastuzumab. Inset numbers indicate the percentage of active caspase 3þ SKBR3 cells. (Continued on the following page.)

    Article Snippet: The human HER2-positive (HER2þ) breast cancer cell line SKBR3 (ATCC catalog no. HTB-30, RRID:CVCL_0033) was obtained from ATCC.

    Techniques: Control

    Figure 7. NK-cell signature, NK-cell effector molecules, CD137 axis, and TGFb expression in treatment-naive breast carcinomas and their association with response to trastuzumab. Levels of the gene signature predicting NK-cell infiltration (see “Materials and Methods”), IFNG, GZMB, CCL5, FCGR3A (CD16), KLRK1 (NKG2D), CXCL9, CXCL10, TGFB1–3, as well as TNFRSF9 (CD137) and TNFSF9 (CD137L) were analyzed in treatment-na€ve HER2þ breast carcinomas from a phase II trial dataset (GSE130786). A, Violin plot showing NK-cell GSVA scores in tumors from patients achieving pCR to trastuzumab neoadjuvant treatment (n ¼ 40) or not (n ¼ 41). TI, tumor-infiltrating. B, Correlation between tumor-infiltrating NK-cell score and GZMB, CCL5, IFNG, and TGFB1–3 scores. Statistical significance calculated by Pearson test. C, Correlation between tumor-infiltrating NK-cell score and TNFRSF9. D, Correlation of TNFRSF9 and TNFSF9 between themselves and with NK-cell effector molecules. Statistical significance calculated by Pearson test. E, Association of CCL5, GZMB, IFNG, TNFRSF9, TNFSF9, KLRK1, FCGR3A, CXCL10, and CXCL9 scores in tumors from patients achieving pCR to trastuzumab or not. Statistical significance by unpaired Student t test. F, OR and 97.5% CI of NK cell, IFNG, GZMB, CCL5, TGFB1, TGFB2, and TGFB3 scores for pCR to trastuzumab.

    Journal: Cancer Immunology Research

    Article Title: CD137 Costimulation Counteracts TGFβ Inhibition of NK-cell Antitumor Function

    doi: 10.1158/2326-6066.cir-21-0030

    Figure Lengend Snippet: Figure 7. NK-cell signature, NK-cell effector molecules, CD137 axis, and TGFb expression in treatment-naive breast carcinomas and their association with response to trastuzumab. Levels of the gene signature predicting NK-cell infiltration (see “Materials and Methods”), IFNG, GZMB, CCL5, FCGR3A (CD16), KLRK1 (NKG2D), CXCL9, CXCL10, TGFB1–3, as well as TNFRSF9 (CD137) and TNFSF9 (CD137L) were analyzed in treatment-na€ve HER2þ breast carcinomas from a phase II trial dataset (GSE130786). A, Violin plot showing NK-cell GSVA scores in tumors from patients achieving pCR to trastuzumab neoadjuvant treatment (n ¼ 40) or not (n ¼ 41). TI, tumor-infiltrating. B, Correlation between tumor-infiltrating NK-cell score and GZMB, CCL5, IFNG, and TGFB1–3 scores. Statistical significance calculated by Pearson test. C, Correlation between tumor-infiltrating NK-cell score and TNFRSF9. D, Correlation of TNFRSF9 and TNFSF9 between themselves and with NK-cell effector molecules. Statistical significance calculated by Pearson test. E, Association of CCL5, GZMB, IFNG, TNFRSF9, TNFSF9, KLRK1, FCGR3A, CXCL10, and CXCL9 scores in tumors from patients achieving pCR to trastuzumab or not. Statistical significance by unpaired Student t test. F, OR and 97.5% CI of NK cell, IFNG, GZMB, CCL5, TGFB1, TGFB2, and TGFB3 scores for pCR to trastuzumab.

    Article Snippet: The human HER2-positive (HER2þ) breast cancer cell line SKBR3 (ATCC catalog no. HTB-30, RRID:CVCL_0033) was obtained from ATCC.

    Techniques: Expressing