mesothelin Search Results


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  • 99
    Thermo Fisher mesothelin specific antibodies
    Mice injected with anti-TIM-3/CD137 mAb developed a tumor antigen-specific CTL response. A , mice injected i.p. with 1 × 10 6 ID8 cells 10 day earlier were injected thrice at 4 days interval with 250 μg of anti-TIM-3/CD137 mAb. Seven days after the last mAb injection, splenocytes from treated mice were cultured in the presence or absence of H-2Db-restricted <t>mesothelin-derived</t> peptides or control HPV-E7-derived peptide for 3 days and IFN-γ production in the supernatants were assayed by ELISA. B , mice were treated as described in (A) . Seven days after the last mAb injection, splenocytes (5 × 10 6 ) from 3 mice were incubated with 5 × 10 5 UV-irradiated ID8 cells for 4 days prior to subject to analysis of antigen-specific CTL activity by CytoTox 96 Non-radioactive cytotoxicity assay using EL4 cells pulsed with H-2Db-restricted mesothelin or HPV-E7 peptide as target cells. The killing activity was also evaluated in the presence of anti-CD4, anti-CD8 or control antibody (C) . Data were expressed as M ± SEM of triplicate wells.
    Mesothelin Specific Antibodies, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 23 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Novocastra mesothelin
    Representative samples of pancreatic ductal adenocarcinoma tissues that were awarded <t>mesothelin</t> expression scores of 0, +1, and +2. (A) Intraductal papillary mucinous adenoma tissue sample (stain, HE). (B) Mesothelin expression was faintly or barely detected in the tumor cell cytoplasm of (A) (mesothelin staining intensity of +1). No luminal membrane staining was detected in the adenoma cells. (C) Invasive intraductal papillary mucinous carcinoma (stain, HE). (D) Moderate to strong mesothelin expression was detected in the carcinoma cells (mesothelin staining intensity of +2). Granular cytoplasmic staining and staining of the entire cell membrane circumference were detected. This case was classified as luminal membrane-positive and cytoplasm-positive. Magnification, ×200. HE, hematoxylin and eosin.
    Mesothelin, supplied by Novocastra, used in various techniques. Bioz Stars score: 91/100, based on 49 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    BioLegend mesothelin
    Mesothelioma cells release GM-CSF to upregulate granulocyte ROS and suppressive activity. A, Cytokine multiplex assay determined the cytokine profile of tumor cell supernatants and cell line supernatants. Increased concentrations of GM-CSF, IL8, GCSF, VEGF, IL6, and <t>mesothelin</t> are found. Low concentrations of prostaglandin E2 and IL13 were detected. B, Transcriptomic expression of GMCSF, GCSF, IL6, IL13, IL8, VEGF, and mesothelin in 87 mesothelioma tumors from the R2: Genomics and Visualisation Platform. C, ROS production (DCFDA staining) by healthy donor CD15 + cells treated with detected cytokines to determine which were capable of enhancing ROS production. GM-CSF increased ROS production most prominently. D, T-cell proliferation was significantly suppressed by granulocytes conditioned with recombinant GM-CSF, compared with control granulocytes. Ratios of 1:1 and 1:0.5 T cells:granulocytes shown. E, Inhibition of granulocyte ROS production (iNAC) or accumulation (catalase) after healthy-donor granulocytes were conditioned with GM-CSF restores T-cell proliferation compared with controls.
    Mesothelin, supplied by BioLegend, used in various techniques. Bioz Stars score: 91/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Aduro mesothelin
    FACS analysis of <t>mesothelin</t> surface expression of AGS cells. Live cells were labeled with anti-mesothelin antibody and then antibody was detected with PE-conjugated secondary antibody. A) Representative histogram of PE detection in greater than 20,000 AGS cells (shaded curve) compared to control (unfilled curve). B) WST-8 cell viability assay of SS1P treated AGS cells. 100% viability was normalized to measurement for untreated cells and 0% viability to cells treated with 10 mcg/mL cyclohexamide which produces complete cell death. A representative cytotoxicity curves is shown. Each point represents the average of triplicate measurements. The dotted horizontal line marks 50% cell killing. The vertical arrow indicates the IC50 for the run. The average IC50 after 8 experiments was 0.4 ng/mL.
    Mesothelin, supplied by Aduro, used in various techniques. Bioz Stars score: 91/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Cell Signaling Technology Inc antibodies against mesothelin
    <t>MUC16</t> expression correlates with <t>MSLN-mediated</t> MMP-7 induction in pancreatic cancer cells. hTERT-HPNE, HPDE, PANC-1, SW1990, and Pa03C cells were starved in serum-free medium and incubated in the presence or absence of MSLN (1 μg/ml) overnight. (A) MMP-7, (B) MMP-2, and (C) MMP-9 mRNA levels were then determined by qRT-PCR. GAPDH served as internal control. Data represent the mean ± S.E. of at least three independent experiments. *, p
    Antibodies Against Mesothelin, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Abcam mesothelin
    <t>Mesothelin</t> silencing reduces proliferation rate of pancreatic and ovarian cancer cells. (A)Mesothelin protein was detected in pancreatic cancer cell lines, Panc1, Miapaca2 and Bxpc3 and ovarian cancer cells Skov3 and Ovcar3. NIH3T3 and Huvec cells which are void of mesothelin were used to prove the specificity of mesothelin antibody. (B)Skov3 cells had reduced proliferation at day 3 post-electroporation with anti-mesothelin siRNA to about 50% of negative control. Callout panels show the density of cell at each time-point. (C–D)Two pancreatic cancer cell lines, Bxpc3 and Miapaca, were tested for the outcome of silencing mesothelin on their proliferation. In both cases a significant loss of proliferation was observed, however for Bxpc3 the decline initiates at later time points as compared with Miapaca cells. For both cells, once again, a rebound to higher proliferation rates is observed at longer time-points due to the clearance of siRNA from cells. Callout panels show the density of cell at each time-point.
    Mesothelin, supplied by Abcam, used in various techniques. Bioz Stars score: 94/100, based on 22 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Thermo Fisher mesothelin
    TNBC frequently expresses <t>mesothelin.</t> 20× magnifications views of light microscopy of stained sections were depicted in a, b, and c. a) mesothelioma positive control; b) TNBC with ~ 5% tumor cells staining positive for mesothelin; c) TNBC with
    Mesothelin, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 91/100, based on 141 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Novus Biologicals mesothelin lysate
    Serum levels of <t>anti-mesothelin</t> antibody increases after treatment. Pre- and post-treatment sera were collected from 5 patients (3 lung cancer, 1 ovarian, 1 pancreatic) and screened for antibodies against mesothelin by ELISA. Data represent the percent increase in OD 450nm units of post-treatment sera over the corresponding pre-treatment sera values.
    Mesothelin Lysate, supplied by Novus Biologicals, used in various techniques. Bioz Stars score: 85/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    R&D Systems mesothelin antibody
    Serum levels of <t>anti-mesothelin</t> antibody increases after treatment. Pre- and post-treatment sera were collected from 5 patients (3 lung cancer, 1 ovarian, 1 pancreatic) and screened for antibodies against mesothelin by ELISA. Data represent the percent increase in OD 450nm units of post-treatment sera over the corresponding pre-treatment sera values.
    Mesothelin Antibody, supplied by R&D Systems, used in various techniques. Bioz Stars score: 99/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Santa Cruz Biotechnology mesothelin
    Activation of NOX4 and <t>mesothelin</t> deposition in a BCG-induced pleurisy mouse model. NOX4 and mesothelin (markers for PMCs) were subjected to immunofluorescence staining. A1 – D1 : DAPI staining. A2 – D2 : Immunofluorescence staining of mesothelin (green color). A3 – D3 : Immunofluorescence staining of NOX4 (red color). A4 – D4 : Overlays of immunofluorescence staining and DAPI staining, yellow areas represent colocalization of mesothelin and NOX4. DAPI, 4′,6-Diamidino-2-phenylindole dihydrochloride.
    Mesothelin, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 92/100, based on 86 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    R&D Systems mesothelin
    CD26 high cells are cytotoxic and polyfunctional in vitro when engineered with a chimeric antigen receptor. A ) Transduction method. αCD3/ICOS-stimulated CD4 + T cell subsets were genetically engineered with a 1 st generation <t>mesothelin-specific</t> CAR. Cells were expanded for 6 days and analyzed by flow cytometry for CAR expression prior to use. B ) Percentage of K562-meso cells that were lysed by effector CD4 + T cell subsets. C ) Cytokine secretion determined by ELISA. Representative of 3 experiments.
    Mesothelin, supplied by R&D Systems, used in various techniques. Bioz Stars score: 91/100, based on 17 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Novocastra anti mesothelin
    KLF8 alone is sufficient to induce T80 cells to form subcutaneous tumors resulting shortened life. A , Representative subcutaneous tumors formed by the indicated cell lines. Mock and SKOV3-ip1 were used as negative and positive controls, respectively. Tumorigenesis experiments were carried out as described in Materials and Methods. Photos were taken on 30 days (for SKOV3-ip1) or 105 days after injection. B , Tumor formation rates Tumor volumes were recorded at the indicated time points (see tumor incidence in Supplemental Table 1 ). C , T80/KLF8 tumor formation shortened mouse survival time. Survival distribution was presented by Kaplan-Meier curve. D , The T80/KLF8 tumors are highly similar to tumors of ovarian cancer patients. H E and IHC staining of the tumors were performed as described in Materials and Methods for expression of the KLF8 and human ovarian cancer marker proteins. a, H E (100X); b, KLF8 (100X); c, pan-cytokeratins (100X); d, CA 125 (100X); e, <t>mesothelin</t> (100X); f, HE4 (100X). Inlets, 400X
    Anti Mesothelin, supplied by Novocastra, used in various techniques. Bioz Stars score: 88/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Roche human mesothelin
    KLF8 alone is sufficient to induce T80 cells to form subcutaneous tumors resulting shortened life. A , Representative subcutaneous tumors formed by the indicated cell lines. Mock and SKOV3-ip1 were used as negative and positive controls, respectively. Tumorigenesis experiments were carried out as described in Materials and Methods. Photos were taken on 30 days (for SKOV3-ip1) or 105 days after injection. B , Tumor formation rates Tumor volumes were recorded at the indicated time points (see tumor incidence in Supplemental Table 1 ). C , T80/KLF8 tumor formation shortened mouse survival time. Survival distribution was presented by Kaplan-Meier curve. D , The T80/KLF8 tumors are highly similar to tumors of ovarian cancer patients. H E and IHC staining of the tumors were performed as described in Materials and Methods for expression of the KLF8 and human ovarian cancer marker proteins. a, H E (100X); b, KLF8 (100X); c, pan-cytokeratins (100X); d, CA 125 (100X); e, <t>mesothelin</t> (100X); f, HE4 (100X). Inlets, 400X
    Human Mesothelin, supplied by Roche, used in various techniques. Bioz Stars score: 93/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Fujirebio mesothelin
    Scatterplot of calretinin versus <t>mesothelin.</t> The plot shows marker concentrations of MM cases and controls from Australia (group 2) and Germany (group 3)
    Mesothelin, supplied by Fujirebio, used in various techniques. Bioz Stars score: 90/100, based on 31 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Genentech mesothelin
    Immunohistochemical staining for <t>mesothelin</t> of HPAC tumor material from mice injected with 100 μg AMA-800CW (tumor harvested 144 h after injection) A. A representative brightfield image (400x) of a tumor with an IHC staining for mesothelin. B. Fluorescence microscopy at 800 nm for IRDye 800CW labelled to AMA; C. at 460 nm for DAPI staining, D. and overlay of fluorescent images.
    Mesothelin, supplied by Genentech, used in various techniques. Bioz Stars score: 91/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Leica Microsystems mesothelin
    Immunohistochemical staining for <t>mesothelin</t> of HPAC tumor material from mice injected with 100 μg AMA-800CW (tumor harvested 144 h after injection) A. A representative brightfield image (400x) of a tumor with an IHC staining for mesothelin. B. Fluorescence microscopy at 800 nm for IRDye 800CW labelled to AMA; C. at 460 nm for DAPI staining, D. and overlay of fluorescent images.
    Mesothelin, supplied by Leica Microsystems, used in various techniques. Bioz Stars score: 91/100, based on 27 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    peptides&elephants mesothelin peptide pool
    Whole-blood IFN-γ responses to peptides spanning the MPF and mature <t>mesothelin</t> components 42 non-overlapping 15-mer peptides spanning the entire mesothelin molecules were exposed to peripheral blood of GBM patients over a seven-day period. Supernatants were then harvested for IFN-γ detection by ELISA. Absolute IFN-γ concentrations (pg/ml) produced by each patient for a single peptide A. as well as the average value per peptide (total IFN-γ/no. of patients) normalised to the sum of IFN-γ production for the entire peptide pool in percentage B. are shown. Immune hotspots within the mesothelin component, defined by peptide-specific IFN-γ production, identify several peptides which may represent viable targets to expand T-cells in host-directed therapies.
    Mesothelin Peptide Pool, supplied by peptides&elephants, used in various techniques. Bioz Stars score: 91/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Signet Testing mesothelin
    Whole-blood IFN-γ responses to peptides spanning the MPF and mature <t>mesothelin</t> components 42 non-overlapping 15-mer peptides spanning the entire mesothelin molecules were exposed to peripheral blood of GBM patients over a seven-day period. Supernatants were then harvested for IFN-γ detection by ELISA. Absolute IFN-γ concentrations (pg/ml) produced by each patient for a single peptide A. as well as the average value per peptide (total IFN-γ/no. of patients) normalised to the sum of IFN-γ production for the entire peptide pool in percentage B. are shown. Immune hotspots within the mesothelin component, defined by peptide-specific IFN-γ production, identify several peptides which may represent viable targets to expand T-cells in host-directed therapies.
    Mesothelin, supplied by Signet Testing, used in various techniques. Bioz Stars score: 91/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Immuno-Biological Laboratories Co Ltd mesothelin
    Plasma endpoint biomarkers. Rats were injected with 100 μL serum free media or serum free media containing 5 × 10 5 parental or chemo-resistant II-45 cells directly into the pleural cavity. At ethical endpoint, rats were euthanized and plasma samples were analysed for cytokine levels relative to rats with parental II-45 mesothelioma. (a) osteopontin levels; (b) <t>mesothelin</t> levels. Normal control rats, Control; parental mesothelioma cells, II-45; cisplatin resistant II-45 cells, CisR; pemetrexed resistant II-45 cells, PemR; combination (cisplatin + pemetrexed) resistant II-45 cells, ComboR; gemcitabine resistant II-45 cells, GemR; vinorelbine resistant II-45 cells, VLBR. P-values were calculated using a one-way Anova test with a value of less than 0.05 indicating significance. * p
    Mesothelin, supplied by Immuno-Biological Laboratories Co Ltd, used in various techniques. Bioz Stars score: 91/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Morphotek mesothelin
    Plasma endpoint biomarkers. Rats were injected with 100 μL serum free media or serum free media containing 5 × 10 5 parental or chemo-resistant II-45 cells directly into the pleural cavity. At ethical endpoint, rats were euthanized and plasma samples were analysed for cytokine levels relative to rats with parental II-45 mesothelioma. (a) osteopontin levels; (b) <t>mesothelin</t> levels. Normal control rats, Control; parental mesothelioma cells, II-45; cisplatin resistant II-45 cells, CisR; pemetrexed resistant II-45 cells, PemR; combination (cisplatin + pemetrexed) resistant II-45 cells, ComboR; gemcitabine resistant II-45 cells, GemR; vinorelbine resistant II-45 cells, VLBR. P-values were calculated using a one-way Anova test with a value of less than 0.05 indicating significance. * p
    Mesothelin, supplied by Morphotek, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    BioScience Inc mesothelin
    Plasma endpoint biomarkers. Rats were injected with 100 μL serum free media or serum free media containing 5 × 10 5 parental or chemo-resistant II-45 cells directly into the pleural cavity. At ethical endpoint, rats were euthanized and plasma samples were analysed for cytokine levels relative to rats with parental II-45 mesothelioma. (a) osteopontin levels; (b) <t>mesothelin</t> levels. Normal control rats, Control; parental mesothelioma cells, II-45; cisplatin resistant II-45 cells, CisR; pemetrexed resistant II-45 cells, PemR; combination (cisplatin + pemetrexed) resistant II-45 cells, ComboR; gemcitabine resistant II-45 cells, GemR; vinorelbine resistant II-45 cells, VLBR. P-values were calculated using a one-way Anova test with a value of less than 0.05 indicating significance. * p
    Mesothelin, supplied by BioScience Inc, used in various techniques. Bioz Stars score: 90/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Mice injected with anti-TIM-3/CD137 mAb developed a tumor antigen-specific CTL response. A , mice injected i.p. with 1 × 10 6 ID8 cells 10 day earlier were injected thrice at 4 days interval with 250 μg of anti-TIM-3/CD137 mAb. Seven days after the last mAb injection, splenocytes from treated mice were cultured in the presence or absence of H-2Db-restricted mesothelin-derived peptides or control HPV-E7-derived peptide for 3 days and IFN-γ production in the supernatants were assayed by ELISA. B , mice were treated as described in (A) . Seven days after the last mAb injection, splenocytes (5 × 10 6 ) from 3 mice were incubated with 5 × 10 5 UV-irradiated ID8 cells for 4 days prior to subject to analysis of antigen-specific CTL activity by CytoTox 96 Non-radioactive cytotoxicity assay using EL4 cells pulsed with H-2Db-restricted mesothelin or HPV-E7 peptide as target cells. The killing activity was also evaluated in the presence of anti-CD4, anti-CD8 or control antibody (C) . Data were expressed as M ± SEM of triplicate wells.

    Journal: Journal of Translational Medicine

    Article Title: Combined TIM-3 blockade and CD137 activation affords the long-term protection in a murine model of ovarian cancer

    doi: 10.1186/1479-5876-11-215

    Figure Lengend Snippet: Mice injected with anti-TIM-3/CD137 mAb developed a tumor antigen-specific CTL response. A , mice injected i.p. with 1 × 10 6 ID8 cells 10 day earlier were injected thrice at 4 days interval with 250 μg of anti-TIM-3/CD137 mAb. Seven days after the last mAb injection, splenocytes from treated mice were cultured in the presence or absence of H-2Db-restricted mesothelin-derived peptides or control HPV-E7-derived peptide for 3 days and IFN-γ production in the supernatants were assayed by ELISA. B , mice were treated as described in (A) . Seven days after the last mAb injection, splenocytes (5 × 10 6 ) from 3 mice were incubated with 5 × 10 5 UV-irradiated ID8 cells for 4 days prior to subject to analysis of antigen-specific CTL activity by CytoTox 96 Non-radioactive cytotoxicity assay using EL4 cells pulsed with H-2Db-restricted mesothelin or HPV-E7 peptide as target cells. The killing activity was also evaluated in the presence of anti-CD4, anti-CD8 or control antibody (C) . Data were expressed as M ± SEM of triplicate wells.

    Article Snippet: The presence of mesothelin-specific antibodies was determined by staining the mouse ID8 ovarian cancer cells using serum from treated mice in a 1:200 dilution, followed by Phycoerythrin-conjugated anti-mouse IgG antibody (eBioscience) staining.

    Techniques: Mouse Assay, Injection, CTL Assay, Cell Culture, Derivative Assay, Enzyme-linked Immunosorbent Assay, Incubation, Irradiation, Activity Assay, Cytotoxicity Assay

    Representative samples of pancreatic ductal adenocarcinoma tissues that were awarded mesothelin expression scores of 0, +1, and +2. (A) Intraductal papillary mucinous adenoma tissue sample (stain, HE). (B) Mesothelin expression was faintly or barely detected in the tumor cell cytoplasm of (A) (mesothelin staining intensity of +1). No luminal membrane staining was detected in the adenoma cells. (C) Invasive intraductal papillary mucinous carcinoma (stain, HE). (D) Moderate to strong mesothelin expression was detected in the carcinoma cells (mesothelin staining intensity of +2). Granular cytoplasmic staining and staining of the entire cell membrane circumference were detected. This case was classified as luminal membrane-positive and cytoplasm-positive. Magnification, ×200. HE, hematoxylin and eosin.

    Journal: Oncology Letters

    Article Title: Importance of luminal membrane mesothelin expression in intraductal papillary mucinous neoplasms

    doi: 10.3892/ol.2015.2969

    Figure Lengend Snippet: Representative samples of pancreatic ductal adenocarcinoma tissues that were awarded mesothelin expression scores of 0, +1, and +2. (A) Intraductal papillary mucinous adenoma tissue sample (stain, HE). (B) Mesothelin expression was faintly or barely detected in the tumor cell cytoplasm of (A) (mesothelin staining intensity of +1). No luminal membrane staining was detected in the adenoma cells. (C) Invasive intraductal papillary mucinous carcinoma (stain, HE). (D) Moderate to strong mesothelin expression was detected in the carcinoma cells (mesothelin staining intensity of +2). Granular cytoplasmic staining and staining of the entire cell membrane circumference were detected. This case was classified as luminal membrane-positive and cytoplasm-positive. Magnification, ×200. HE, hematoxylin and eosin.

    Article Snippet: Subsequently, the slides were incubated with a 1:50 dilution of a mouse monoclonal antibody for mesothelin (clone 5B2; Novocastra, Newcastle-Upon-Tyne, United Kingdom) at room temperature for 30 min.

    Techniques: Expressing, Staining

    Flow chart of mesothelin expression in IPMN cells. IPMN, intraductal papillary mucinous neoplasm.

    Journal: Oncology Letters

    Article Title: Importance of luminal membrane mesothelin expression in intraductal papillary mucinous neoplasms

    doi: 10.3892/ol.2015.2969

    Figure Lengend Snippet: Flow chart of mesothelin expression in IPMN cells. IPMN, intraductal papillary mucinous neoplasm.

    Article Snippet: Subsequently, the slides were incubated with a 1:50 dilution of a mouse monoclonal antibody for mesothelin (clone 5B2; Novocastra, Newcastle-Upon-Tyne, United Kingdom) at room temperature for 30 min.

    Techniques: Flow Cytometry, Expressing

    Mesothelin expression was detected in IPMN tissue, but not in the normal pancreatic tissue, and was limited to adenoma cells (arrows) Notably, the benign ductal epithelial cells of the intraductal papillary mucinous adenoma patients were negative for mesothelin expression (arrowheads). (A) Stain, hematoxylin and eosin (HE). (B) Tissue sample in (A) stained for mesothelin expression. (C) Stain, HE. (D) Tissue sample in (C) stained for mesothelin expression. Magnification, ×200.

    Journal: Oncology Letters

    Article Title: Importance of luminal membrane mesothelin expression in intraductal papillary mucinous neoplasms

    doi: 10.3892/ol.2015.2969

    Figure Lengend Snippet: Mesothelin expression was detected in IPMN tissue, but not in the normal pancreatic tissue, and was limited to adenoma cells (arrows) Notably, the benign ductal epithelial cells of the intraductal papillary mucinous adenoma patients were negative for mesothelin expression (arrowheads). (A) Stain, hematoxylin and eosin (HE). (B) Tissue sample in (A) stained for mesothelin expression. (C) Stain, HE. (D) Tissue sample in (C) stained for mesothelin expression. Magnification, ×200.

    Article Snippet: Subsequently, the slides were incubated with a 1:50 dilution of a mouse monoclonal antibody for mesothelin (clone 5B2; Novocastra, Newcastle-Upon-Tyne, United Kingdom) at room temperature for 30 min.

    Techniques: Expressing, Staining

    Mesothelioma cells release GM-CSF to upregulate granulocyte ROS and suppressive activity. A, Cytokine multiplex assay determined the cytokine profile of tumor cell supernatants and cell line supernatants. Increased concentrations of GM-CSF, IL8, GCSF, VEGF, IL6, and mesothelin are found. Low concentrations of prostaglandin E2 and IL13 were detected. B, Transcriptomic expression of GMCSF, GCSF, IL6, IL13, IL8, VEGF, and mesothelin in 87 mesothelioma tumors from the R2: Genomics and Visualisation Platform. C, ROS production (DCFDA staining) by healthy donor CD15 + cells treated with detected cytokines to determine which were capable of enhancing ROS production. GM-CSF increased ROS production most prominently. D, T-cell proliferation was significantly suppressed by granulocytes conditioned with recombinant GM-CSF, compared with control granulocytes. Ratios of 1:1 and 1:0.5 T cells:granulocytes shown. E, Inhibition of granulocyte ROS production (iNAC) or accumulation (catalase) after healthy-donor granulocytes were conditioned with GM-CSF restores T-cell proliferation compared with controls.

    Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

    Article Title: Tumor-Derived GM-CSF Promotes Granulocyte Immunosuppression in Mesothelioma Patients

    doi: 10.1158/1078-0432.CCR-17-3757

    Figure Lengend Snippet: Mesothelioma cells release GM-CSF to upregulate granulocyte ROS and suppressive activity. A, Cytokine multiplex assay determined the cytokine profile of tumor cell supernatants and cell line supernatants. Increased concentrations of GM-CSF, IL8, GCSF, VEGF, IL6, and mesothelin are found. Low concentrations of prostaglandin E2 and IL13 were detected. B, Transcriptomic expression of GMCSF, GCSF, IL6, IL13, IL8, VEGF, and mesothelin in 87 mesothelioma tumors from the R2: Genomics and Visualisation Platform. C, ROS production (DCFDA staining) by healthy donor CD15 + cells treated with detected cytokines to determine which were capable of enhancing ROS production. GM-CSF increased ROS production most prominently. D, T-cell proliferation was significantly suppressed by granulocytes conditioned with recombinant GM-CSF, compared with control granulocytes. Ratios of 1:1 and 1:0.5 T cells:granulocytes shown. E, Inhibition of granulocyte ROS production (iNAC) or accumulation (catalase) after healthy-donor granulocytes were conditioned with GM-CSF restores T-cell proliferation compared with controls.

    Article Snippet: The following molecules were tested GM-CSF (Biolegend), IL13 (BD Biosciences), IL8 (BioLegend), IL6 (BioLegend), G-CSF (R & D Systems), VEGF (R & D Systems), Mesothelin (BioLegend).

    Techniques: Activity Assay, Multiplex Assay, Expressing, Staining, Recombinant, Inhibition

    FACS analysis of mesothelin surface expression of AGS cells. Live cells were labeled with anti-mesothelin antibody and then antibody was detected with PE-conjugated secondary antibody. A) Representative histogram of PE detection in greater than 20,000 AGS cells (shaded curve) compared to control (unfilled curve). B) WST-8 cell viability assay of SS1P treated AGS cells. 100% viability was normalized to measurement for untreated cells and 0% viability to cells treated with 10 mcg/mL cyclohexamide which produces complete cell death. A representative cytotoxicity curves is shown. Each point represents the average of triplicate measurements. The dotted horizontal line marks 50% cell killing. The vertical arrow indicates the IC50 for the run. The average IC50 after 8 experiments was 0.4 ng/mL.

    Journal: Applied immunohistochemistry & molecular morphology : AIMM / official publication of the Society for Applied Immunohistochemistry

    Article Title: Mesothelin expression in advanced gastro-esophageal cancer represents a novel target for immunotherapy

    doi: 10.1097/PAI.0000000000000292

    Figure Lengend Snippet: FACS analysis of mesothelin surface expression of AGS cells. Live cells were labeled with anti-mesothelin antibody and then antibody was detected with PE-conjugated secondary antibody. A) Representative histogram of PE detection in greater than 20,000 AGS cells (shaded curve) compared to control (unfilled curve). B) WST-8 cell viability assay of SS1P treated AGS cells. 100% viability was normalized to measurement for untreated cells and 0% viability to cells treated with 10 mcg/mL cyclohexamide which produces complete cell death. A representative cytotoxicity curves is shown. Each point represents the average of triplicate measurements. The dotted horizontal line marks 50% cell killing. The vertical arrow indicates the IC50 for the run. The average IC50 after 8 experiments was 0.4 ng/mL.

    Article Snippet: IHC for mesothelin (clone 2C6, Aduro BioTech, Inc., Berkeley, CA) was performed using a previously standardized protocol (unpublished) on a Leica Bond autostainer (Leica Biosystems, Buffalo Grove, IL) with a biotin free polymer detection (Bond Polymer Refine Detection, Leica Biosystems, Buffalo Grove, IL).

    Techniques: FACS, Expressing, Labeling, Viability Assay

    MUC16 expression correlates with MSLN-mediated MMP-7 induction in pancreatic cancer cells. hTERT-HPNE, HPDE, PANC-1, SW1990, and Pa03C cells were starved in serum-free medium and incubated in the presence or absence of MSLN (1 μg/ml) overnight. (A) MMP-7, (B) MMP-2, and (C) MMP-9 mRNA levels were then determined by qRT-PCR. GAPDH served as internal control. Data represent the mean ± S.E. of at least three independent experiments. *, p

    Journal: Scientific Reports

    Article Title: Mesothelin Binding to CA125/MUC16 Promotes Pancreatic Cancer Cell Motility and Invasion via MMP-7 Activation

    doi: 10.1038/srep01870

    Figure Lengend Snippet: MUC16 expression correlates with MSLN-mediated MMP-7 induction in pancreatic cancer cells. hTERT-HPNE, HPDE, PANC-1, SW1990, and Pa03C cells were starved in serum-free medium and incubated in the presence or absence of MSLN (1 μg/ml) overnight. (A) MMP-7, (B) MMP-2, and (C) MMP-9 mRNA levels were then determined by qRT-PCR. GAPDH served as internal control. Data represent the mean ± S.E. of at least three independent experiments. *, p

    Article Snippet: Western blotting Western blotting assays were performed as previously described and stained with antibodies for MUC16, MSLN, MMP-7, MMP-9, β-actin, or signaling proteins including ERK1/2, p-ERK1/2 (Thr202/Tyr204), Akt, p-Akt (Ser473), p38 MAPK, and p-p38 MAPK (Thr180/Tyr182) (Cell Signaling Technology, Danvers, MA).

    Techniques: Expressing, Incubation, Quantitative RT-PCR

    Proposed molecular pathway induced by MSLN-MUC16 binding, which results in MMP-7 induction in pancreatic cancer cells. The binding of MSLN to MUC16 present on the pancreatic cancer cell surface activates a p38 MAPK-dependent pathway, which in turn upregulates MMP-7 synthesis, resulting in increased invasive and migratory potentials. In the absence of MUC16 expression by pancreatic cancer cells, MSLN is capable of upregulating MMP-7 expression via activation of an ERK-dependent pathway.

    Journal: Scientific Reports

    Article Title: Mesothelin Binding to CA125/MUC16 Promotes Pancreatic Cancer Cell Motility and Invasion via MMP-7 Activation

    doi: 10.1038/srep01870

    Figure Lengend Snippet: Proposed molecular pathway induced by MSLN-MUC16 binding, which results in MMP-7 induction in pancreatic cancer cells. The binding of MSLN to MUC16 present on the pancreatic cancer cell surface activates a p38 MAPK-dependent pathway, which in turn upregulates MMP-7 synthesis, resulting in increased invasive and migratory potentials. In the absence of MUC16 expression by pancreatic cancer cells, MSLN is capable of upregulating MMP-7 expression via activation of an ERK-dependent pathway.

    Article Snippet: Western blotting Western blotting assays were performed as previously described and stained with antibodies for MUC16, MSLN, MMP-7, MMP-9, β-actin, or signaling proteins including ERK1/2, p-ERK1/2 (Thr202/Tyr204), Akt, p-Akt (Ser473), p38 MAPK, and p-p38 MAPK (Thr180/Tyr182) (Cell Signaling Technology, Danvers, MA).

    Techniques: Binding Assay, Expressing, Activation Assay

    MSLN and MUC16 co-immunoprecipitate in pancreatic cancer cells. Whole lysates from scramble control (A) or MUC16-KD (B) SW1990 and Pa03C cells were immunoprecipitated with either an anti-MUC16 or anti-MSLN or an isotype control antibody, and analyzed for MSLN or MUC16 reactivity by immunoblotting. Lysates from SW1990 and Pa03C cells were also analyzed as input controls.

    Journal: Scientific Reports

    Article Title: Mesothelin Binding to CA125/MUC16 Promotes Pancreatic Cancer Cell Motility and Invasion via MMP-7 Activation

    doi: 10.1038/srep01870

    Figure Lengend Snippet: MSLN and MUC16 co-immunoprecipitate in pancreatic cancer cells. Whole lysates from scramble control (A) or MUC16-KD (B) SW1990 and Pa03C cells were immunoprecipitated with either an anti-MUC16 or anti-MSLN or an isotype control antibody, and analyzed for MSLN or MUC16 reactivity by immunoblotting. Lysates from SW1990 and Pa03C cells were also analyzed as input controls.

    Article Snippet: Western blotting Western blotting assays were performed as previously described and stained with antibodies for MUC16, MSLN, MMP-7, MMP-9, β-actin, or signaling proteins including ERK1/2, p-ERK1/2 (Thr202/Tyr204), Akt, p-Akt (Ser473), p38 MAPK, and p-p38 MAPK (Thr180/Tyr182) (Cell Signaling Technology, Danvers, MA).

    Techniques: Immunoprecipitation

    MSLN-MUC16 interaction promotes pancreatic cancer cell invasion via MMP-7 activation. Scramble control or MUC16-KD SW1990 (A) or Pa03C (B) cells were incubated with MSLN (1 μg/ml) in the presence of either a neutralizing MMP-7 antibody (2 μg/ml) or an isotype control and then used in transwell invasion assays. Data are reported as normalized cell number that invaded through the transwell membrane relative to untreated scramble control (100%), and represent the mean ± S.E. of three independent experiments. **, p

    Journal: Scientific Reports

    Article Title: Mesothelin Binding to CA125/MUC16 Promotes Pancreatic Cancer Cell Motility and Invasion via MMP-7 Activation

    doi: 10.1038/srep01870

    Figure Lengend Snippet: MSLN-MUC16 interaction promotes pancreatic cancer cell invasion via MMP-7 activation. Scramble control or MUC16-KD SW1990 (A) or Pa03C (B) cells were incubated with MSLN (1 μg/ml) in the presence of either a neutralizing MMP-7 antibody (2 μg/ml) or an isotype control and then used in transwell invasion assays. Data are reported as normalized cell number that invaded through the transwell membrane relative to untreated scramble control (100%), and represent the mean ± S.E. of three independent experiments. **, p

    Article Snippet: Western blotting Western blotting assays were performed as previously described and stained with antibodies for MUC16, MSLN, MMP-7, MMP-9, β-actin, or signaling proteins including ERK1/2, p-ERK1/2 (Thr202/Tyr204), Akt, p-Akt (Ser473), p38 MAPK, and p-p38 MAPK (Thr180/Tyr182) (Cell Signaling Technology, Danvers, MA).

    Techniques: Activation Assay, Incubation

    MSLN-MUC16 interaction induces MMP-7 synthesis in pancreatic cancer cells. (A) SW1990 cells were starved in serum-free medium overnight and incubated with MSLN (1 μg/ml) for the indicated periods of time. The mRNA levels of MMP-7 were quantified by qRT-PCR. GAPDH served as internal control. Data represent the mean ± S.E. of at least three independent experiments. *, p

    Journal: Scientific Reports

    Article Title: Mesothelin Binding to CA125/MUC16 Promotes Pancreatic Cancer Cell Motility and Invasion via MMP-7 Activation

    doi: 10.1038/srep01870

    Figure Lengend Snippet: MSLN-MUC16 interaction induces MMP-7 synthesis in pancreatic cancer cells. (A) SW1990 cells were starved in serum-free medium overnight and incubated with MSLN (1 μg/ml) for the indicated periods of time. The mRNA levels of MMP-7 were quantified by qRT-PCR. GAPDH served as internal control. Data represent the mean ± S.E. of at least three independent experiments. *, p

    Article Snippet: Western blotting Western blotting assays were performed as previously described and stained with antibodies for MUC16, MSLN, MMP-7, MMP-9, β-actin, or signaling proteins including ERK1/2, p-ERK1/2 (Thr202/Tyr204), Akt, p-Akt (Ser473), p38 MAPK, and p-p38 MAPK (Thr180/Tyr182) (Cell Signaling Technology, Danvers, MA).

    Techniques: Incubation, Quantitative RT-PCR

    MSLN binding to MUC16 promotes pancreatic cancer cell motility via MMP-7 activation. Representative time-lapse images of scramble control and MUC16-KD SW1990 (A) and Pa03C (C) pancreatic cancer cells migrating through channels of prescribed dimensions ( W x H x L = 6 × 10 × 200 μm) using 10% FBS as chemoattractant. In all experiments, cells were incubated with or without MSLN (1 μg/ml) for 6 h before being seeded to the microchannel device. The average migration speed of individual scramble control and MUC16-KD SW1990 (B) and Pa03C (D) cells was determined over a 10 h period for > 30 cells from three independent experiments. *, p

    Journal: Scientific Reports

    Article Title: Mesothelin Binding to CA125/MUC16 Promotes Pancreatic Cancer Cell Motility and Invasion via MMP-7 Activation

    doi: 10.1038/srep01870

    Figure Lengend Snippet: MSLN binding to MUC16 promotes pancreatic cancer cell motility via MMP-7 activation. Representative time-lapse images of scramble control and MUC16-KD SW1990 (A) and Pa03C (C) pancreatic cancer cells migrating through channels of prescribed dimensions ( W x H x L = 6 × 10 × 200 μm) using 10% FBS as chemoattractant. In all experiments, cells were incubated with or without MSLN (1 μg/ml) for 6 h before being seeded to the microchannel device. The average migration speed of individual scramble control and MUC16-KD SW1990 (B) and Pa03C (D) cells was determined over a 10 h period for > 30 cells from three independent experiments. *, p

    Article Snippet: Western blotting Western blotting assays were performed as previously described and stained with antibodies for MUC16, MSLN, MMP-7, MMP-9, β-actin, or signaling proteins including ERK1/2, p-ERK1/2 (Thr202/Tyr204), Akt, p-Akt (Ser473), p38 MAPK, and p-p38 MAPK (Thr180/Tyr182) (Cell Signaling Technology, Danvers, MA).

    Techniques: Binding Assay, Activation Assay, Incubation, Migration

    MUC16 expression correlates with pancreatic cancer cell binding capacity to MSLN. (A) hTERT-HPNE, HPDE, PANC-1, SW1990, and Pa03C cells were seeded in 96-well plates pre-coated with either mesothelin (MSLN) or BSA (control). After 30 min of incubation, the relative number of adherent cells to MSLN- versus BSA-coated wells was quantified by WST-1 assay. Data represent the mean ± S.E. of three independent experiments. *, p

    Journal: Scientific Reports

    Article Title: Mesothelin Binding to CA125/MUC16 Promotes Pancreatic Cancer Cell Motility and Invasion via MMP-7 Activation

    doi: 10.1038/srep01870

    Figure Lengend Snippet: MUC16 expression correlates with pancreatic cancer cell binding capacity to MSLN. (A) hTERT-HPNE, HPDE, PANC-1, SW1990, and Pa03C cells were seeded in 96-well plates pre-coated with either mesothelin (MSLN) or BSA (control). After 30 min of incubation, the relative number of adherent cells to MSLN- versus BSA-coated wells was quantified by WST-1 assay. Data represent the mean ± S.E. of three independent experiments. *, p

    Article Snippet: Western blotting Western blotting assays were performed as previously described and stained with antibodies for MUC16, MSLN, MMP-7, MMP-9, β-actin, or signaling proteins including ERK1/2, p-ERK1/2 (Thr202/Tyr204), Akt, p-Akt (Ser473), p38 MAPK, and p-p38 MAPK (Thr180/Tyr182) (Cell Signaling Technology, Danvers, MA).

    Techniques: Expressing, Binding Assay, Incubation, WST-1 Assay

    MSLN-MUC16 interaction promotes pancreatic cancer cell motility and invasion via MMP-7 induced by a p38 MAPK-dependent pathway. (A) Scramble control and MUC16-KD SW1990 cells were serum-starved overnight, and then treated with MSLN (1 μg/ml) for 12 h. The levels of phospho-ERK1/2 (Thr202/Tyr204) and phospho-p38 MAPK (Thr180/Tyr182) from whole cell lysates were analyzed by immunoblotting using specific Abs. Equal loading in each lane was ensured by the similar intensities of total ERK1/2, p38 MAPK, and β-actin. (B) SW1990 cells were incubated with MSLN (1 μg/ml) for 12 h in the presence or absence of the p38 MAPK inhibitor SB203580 (20 μM). The levels of phospho-p38 MAPK (Thr180/Tyr182) and MMP-7 expression from cell lysates were analyzed by Western blotting using specific Abs. Equal loading in each lane is ensured by the similar intensities of total p38 MAPK and β-actin. These Western blots are representative of three independent experiments, all revealing similar results. The intensity of bands was quantified using the NIH ImageJ software and then normalized with respect to the value obtained for the untreated control. (C, D) SW1990 cells were stimulated with MSLN (1 μg/ml) in the presence or absence of SB203580 (20 μM) or an MMP-7 antisense oligonucleotide (50 nM), and then subjected to either transwell invasion or microchannel motility assays. (C) Data are reported as percentage of untreated control cells that invaded through the transwell membrane, and represent the mean ± S.E. of three independent experiments. (D) The average migration speed of individual cells was determined over a 10 h period for > 30 cells from three independent experiments for each of the indicated conditions. *, p

    Journal: Scientific Reports

    Article Title: Mesothelin Binding to CA125/MUC16 Promotes Pancreatic Cancer Cell Motility and Invasion via MMP-7 Activation

    doi: 10.1038/srep01870

    Figure Lengend Snippet: MSLN-MUC16 interaction promotes pancreatic cancer cell motility and invasion via MMP-7 induced by a p38 MAPK-dependent pathway. (A) Scramble control and MUC16-KD SW1990 cells were serum-starved overnight, and then treated with MSLN (1 μg/ml) for 12 h. The levels of phospho-ERK1/2 (Thr202/Tyr204) and phospho-p38 MAPK (Thr180/Tyr182) from whole cell lysates were analyzed by immunoblotting using specific Abs. Equal loading in each lane was ensured by the similar intensities of total ERK1/2, p38 MAPK, and β-actin. (B) SW1990 cells were incubated with MSLN (1 μg/ml) for 12 h in the presence or absence of the p38 MAPK inhibitor SB203580 (20 μM). The levels of phospho-p38 MAPK (Thr180/Tyr182) and MMP-7 expression from cell lysates were analyzed by Western blotting using specific Abs. Equal loading in each lane is ensured by the similar intensities of total p38 MAPK and β-actin. These Western blots are representative of three independent experiments, all revealing similar results. The intensity of bands was quantified using the NIH ImageJ software and then normalized with respect to the value obtained for the untreated control. (C, D) SW1990 cells were stimulated with MSLN (1 μg/ml) in the presence or absence of SB203580 (20 μM) or an MMP-7 antisense oligonucleotide (50 nM), and then subjected to either transwell invasion or microchannel motility assays. (C) Data are reported as percentage of untreated control cells that invaded through the transwell membrane, and represent the mean ± S.E. of three independent experiments. (D) The average migration speed of individual cells was determined over a 10 h period for > 30 cells from three independent experiments for each of the indicated conditions. *, p

    Article Snippet: Western blotting Western blotting assays were performed as previously described and stained with antibodies for MUC16, MSLN, MMP-7, MMP-9, β-actin, or signaling proteins including ERK1/2, p-ERK1/2 (Thr202/Tyr204), Akt, p-Akt (Ser473), p38 MAPK, and p-p38 MAPK (Thr180/Tyr182) (Cell Signaling Technology, Danvers, MA).

    Techniques: Incubation, Expressing, Western Blot, Software, Migration

    Mesothelin silencing reduces proliferation rate of pancreatic and ovarian cancer cells. (A)Mesothelin protein was detected in pancreatic cancer cell lines, Panc1, Miapaca2 and Bxpc3 and ovarian cancer cells Skov3 and Ovcar3. NIH3T3 and Huvec cells which are void of mesothelin were used to prove the specificity of mesothelin antibody. (B)Skov3 cells had reduced proliferation at day 3 post-electroporation with anti-mesothelin siRNA to about 50% of negative control. Callout panels show the density of cell at each time-point. (C–D)Two pancreatic cancer cell lines, Bxpc3 and Miapaca, were tested for the outcome of silencing mesothelin on their proliferation. In both cases a significant loss of proliferation was observed, however for Bxpc3 the decline initiates at later time points as compared with Miapaca cells. For both cells, once again, a rebound to higher proliferation rates is observed at longer time-points due to the clearance of siRNA from cells. Callout panels show the density of cell at each time-point.

    Journal: PLoS ONE

    Article Title: Inhibition of Mesothelin as a Novel Strategy for Targeting Cancer Cells

    doi: 10.1371/journal.pone.0033214

    Figure Lengend Snippet: Mesothelin silencing reduces proliferation rate of pancreatic and ovarian cancer cells. (A)Mesothelin protein was detected in pancreatic cancer cell lines, Panc1, Miapaca2 and Bxpc3 and ovarian cancer cells Skov3 and Ovcar3. NIH3T3 and Huvec cells which are void of mesothelin were used to prove the specificity of mesothelin antibody. (B)Skov3 cells had reduced proliferation at day 3 post-electroporation with anti-mesothelin siRNA to about 50% of negative control. Callout panels show the density of cell at each time-point. (C–D)Two pancreatic cancer cell lines, Bxpc3 and Miapaca, were tested for the outcome of silencing mesothelin on their proliferation. In both cases a significant loss of proliferation was observed, however for Bxpc3 the decline initiates at later time points as compared with Miapaca cells. For both cells, once again, a rebound to higher proliferation rates is observed at longer time-points due to the clearance of siRNA from cells. Callout panels show the density of cell at each time-point.

    Article Snippet: The membrane was blocked, washed, and incubated with the different primary antibodies such as mesothelin (Abcam, MA and LS Bio, WA) and β-actin (Cell Signaling Technology, MA), followed by the HRP-conjugated secondary antibody.

    Techniques: Electroporation, Negative Control

    The effects of silencing mesothelin on cancer cell invasiveness, pro-oncogenic cell signaling pathways and cell cycle progression. (A) Once tested in a modified Boyden chamber assay, the invasiveness of H2373 mesothelioma cells is reduced significantly (p

    Journal: PLoS ONE

    Article Title: Inhibition of Mesothelin as a Novel Strategy for Targeting Cancer Cells

    doi: 10.1371/journal.pone.0033214

    Figure Lengend Snippet: The effects of silencing mesothelin on cancer cell invasiveness, pro-oncogenic cell signaling pathways and cell cycle progression. (A) Once tested in a modified Boyden chamber assay, the invasiveness of H2373 mesothelioma cells is reduced significantly (p

    Article Snippet: The membrane was blocked, washed, and incubated with the different primary antibodies such as mesothelin (Abcam, MA and LS Bio, WA) and β-actin (Cell Signaling Technology, MA), followed by the HRP-conjugated secondary antibody.

    Techniques: Modification, Boyden Chamber Assay

    Gene specific silencing of mesothelin reduces proliferation of mesothelioma cells. (A)Anti-mesothelin siRNA was designed to a middle sequence position in mesothelin mRNA. (B)Once electroporated with anti-mesothelin siRNA, the expression levels of mesothelin was significantly reduced in H2373 cells. Negative control siRNA did not cause such reduction. Lower panel shows the results of band-densitometry comparing the intensity of mesothelin expression upon electroporation of H2373 cells with siRNA. (C)Anti-mesothelin siRNA did not affect the expression levels of β-actin, a house-keeping protein, as an evidence for the specificity of this anti-mesothelin siRNA for its target. (D) Proliferation rate of H2373 cells is significantly (p

    Journal: PLoS ONE

    Article Title: Inhibition of Mesothelin as a Novel Strategy for Targeting Cancer Cells

    doi: 10.1371/journal.pone.0033214

    Figure Lengend Snippet: Gene specific silencing of mesothelin reduces proliferation of mesothelioma cells. (A)Anti-mesothelin siRNA was designed to a middle sequence position in mesothelin mRNA. (B)Once electroporated with anti-mesothelin siRNA, the expression levels of mesothelin was significantly reduced in H2373 cells. Negative control siRNA did not cause such reduction. Lower panel shows the results of band-densitometry comparing the intensity of mesothelin expression upon electroporation of H2373 cells with siRNA. (C)Anti-mesothelin siRNA did not affect the expression levels of β-actin, a house-keeping protein, as an evidence for the specificity of this anti-mesothelin siRNA for its target. (D) Proliferation rate of H2373 cells is significantly (p

    Article Snippet: The membrane was blocked, washed, and incubated with the different primary antibodies such as mesothelin (Abcam, MA and LS Bio, WA) and β-actin (Cell Signaling Technology, MA), followed by the HRP-conjugated secondary antibody.

    Techniques: Sequencing, Expressing, Negative Control, Electroporation

    A schematic diagram showing the role of Sulfatase-1 in inhibiting mesothelin expression in Carcinogenesis. Desulfation action of sulfatase-1 on heparin sulfate proteoglycan (HSPG) leads to the inhibition of formation of HSPG-Wnt/β-catenin-Msln complex. This action will inhibits the activation of Msln and leads to inhibition of the signaling pathway involved in cell proliferation, migration, invasion, lymphatic metastasis and cell survival

    Journal: Journal of Cell Communication and Signaling

    Article Title: Sulfatase-1 knockdown promotes in vitro and in vivo aggressive behavior of murine hepatocarcinoma Hca-P cells through up-regulation of mesothelin

    doi: 10.1007/s12079-017-0411-9

    Figure Lengend Snippet: A schematic diagram showing the role of Sulfatase-1 in inhibiting mesothelin expression in Carcinogenesis. Desulfation action of sulfatase-1 on heparin sulfate proteoglycan (HSPG) leads to the inhibition of formation of HSPG-Wnt/β-catenin-Msln complex. This action will inhibits the activation of Msln and leads to inhibition of the signaling pathway involved in cell proliferation, migration, invasion, lymphatic metastasis and cell survival

    Article Snippet: The membranes were blocked in 5% non-fat dried milk for one hour and then probed with monoclonal goat anti- Sulfatase 1 (Abcam, China, 1-3 μg/ml), anti- Mesothelin (Abcam, China, 1 μg/ml) and anti-Gapdh (ZSGB-Bio, China, 1:7500) primary antibodies for 1 h. After washing the membranes six times, rabbit anti-goat secondary antibody was applied to Sulf-1, anti- rabbit secondary antibody applied to Msln and anti-mouse secondary antibody applied on Gapdh for 1 h and the bright bands were captured by Li-Cor Odyssey Infrared Imaging System (Version 3.0 software).

    Techniques: Expressing, Inhibition, Activation Assay, Migration

    A schematic diagram showing the role of Sulfatase-1 in Carcinogenesis Desulfation action of sulfatase-1 on heparin sulfate proteoglycan (HSPG) leads to the inhibition of formation of HSPG-Wnt1 complex. This action will inhibits the secretion of Mesothelin which is induced by Wnt1stimulation. Reduction of Msln secretion leads to inhibition of the signaling pathway involved in cell proliferation, migration, invasion, lymphatics metastasis, and hence promote cell survival.

    Journal: Oncotarget

    Article Title: Overexpression of sulfatase-1 in murine hepatocarcinoma Hca-F cell line downregulates mesothelin and leads to reduction in lymphatic metastasis, both in vitro and in vivo

    doi: 10.18632/oncotarget.11933

    Figure Lengend Snippet: A schematic diagram showing the role of Sulfatase-1 in Carcinogenesis Desulfation action of sulfatase-1 on heparin sulfate proteoglycan (HSPG) leads to the inhibition of formation of HSPG-Wnt1 complex. This action will inhibits the secretion of Mesothelin which is induced by Wnt1stimulation. Reduction of Msln secretion leads to inhibition of the signaling pathway involved in cell proliferation, migration, invasion, lymphatics metastasis, and hence promote cell survival.

    Article Snippet: The membrane was blocked in 5% non-fat dried milk for one hour and then probed with monoclonal goat anti- Sulfatase 1 antibody (Abcam, China, 1-3μg/ml), anti- Mesothelin antibody (Abcam, China, 1μg/ml) and Gapdh (ZSGB-Bio, China, 1:7500) primary antibodies for 1hr.

    Techniques: Inhibition, Migration

    TNBC frequently expresses mesothelin. 20× magnifications views of light microscopy of stained sections were depicted in a, b, and c. a) mesothelioma positive control; b) TNBC with ~ 5% tumor cells staining positive for mesothelin; c) TNBC with

    Journal: Breast cancer research and treatment

    Article Title: Mesothelin, a novel immunotherapy target for triple negative breast cancer

    doi: 10.1007/s10549-012-2018-4

    Figure Lengend Snippet: TNBC frequently expresses mesothelin. 20× magnifications views of light microscopy of stained sections were depicted in a, b, and c. a) mesothelioma positive control; b) TNBC with ~ 5% tumor cells staining positive for mesothelin; c) TNBC with

    Article Snippet: Expression of mesothelin was evaluated on formalin fixed paraffin embedded tissues sections by immunohistochemical (IHC) staining using a mouse monoclonal antibody specific for mesothelin (clone 5B2, 1:100) (Thermo Scientific MS-1320) on fully automated Leica Bond™ instrument using the Bond Polymer Refine Detection System.

    Techniques: Light Microscopy, Staining, Positive Control

    Mesothelin is frequently overexpressed in TNBC

    Journal: Breast cancer research and treatment

    Article Title: Mesothelin, a novel immunotherapy target for triple negative breast cancer

    doi: 10.1007/s10549-012-2018-4

    Figure Lengend Snippet: Mesothelin is frequently overexpressed in TNBC

    Article Snippet: Expression of mesothelin was evaluated on formalin fixed paraffin embedded tissues sections by immunohistochemical (IHC) staining using a mouse monoclonal antibody specific for mesothelin (clone 5B2, 1:100) (Thermo Scientific MS-1320) on fully automated Leica Bond™ instrument using the Bond Polymer Refine Detection System.

    Techniques:

    A) Histograms depicting mesothelin expression in various breast cancer cell lines using EM-meso cells as positive control. B) in vitro cell killing assay to evaluate proportion of tumor cells lysed by mesoCAR T-cells vs. non-transduced T cells on EM-meso

    Journal: Breast cancer research and treatment

    Article Title: Mesothelin, a novel immunotherapy target for triple negative breast cancer

    doi: 10.1007/s10549-012-2018-4

    Figure Lengend Snippet: A) Histograms depicting mesothelin expression in various breast cancer cell lines using EM-meso cells as positive control. B) in vitro cell killing assay to evaluate proportion of tumor cells lysed by mesoCAR T-cells vs. non-transduced T cells on EM-meso

    Article Snippet: Expression of mesothelin was evaluated on formalin fixed paraffin embedded tissues sections by immunohistochemical (IHC) staining using a mouse monoclonal antibody specific for mesothelin (clone 5B2, 1:100) (Thermo Scientific MS-1320) on fully automated Leica Bond™ instrument using the Bond Polymer Refine Detection System.

    Techniques: Expressing, Positive Control, In Vitro

    Serum levels of anti-mesothelin antibody increases after treatment. Pre- and post-treatment sera were collected from 5 patients (3 lung cancer, 1 ovarian, 1 pancreatic) and screened for antibodies against mesothelin by ELISA. Data represent the percent increase in OD 450nm units of post-treatment sera over the corresponding pre-treatment sera values.

    Journal: Cancers

    Article Title: Therapeutic Response in Patients with Advanced Malignancies Treated with Combined Dendritic Cell-Activated T Cell Based Immunotherapy and Intensity-Modulated Radiotherapy

    doi: 10.3390/cancers3022223

    Figure Lengend Snippet: Serum levels of anti-mesothelin antibody increases after treatment. Pre- and post-treatment sera were collected from 5 patients (3 lung cancer, 1 ovarian, 1 pancreatic) and screened for antibodies against mesothelin by ELISA. Data represent the percent increase in OD 450nm units of post-treatment sera over the corresponding pre-treatment sera values.

    Article Snippet: Test wells were coated with 1.0 mcg/mL KLH (Calbiochem, San Diego, CA) in 1 × PBS or 20 mcg/mL mesothelin lysate (Novus Biologicals) in 0.1 M carbonate buffer.

    Techniques: Enzyme-linked Immunosorbent Assay

    Activation of NOX4 and mesothelin deposition in a BCG-induced pleurisy mouse model. NOX4 and mesothelin (markers for PMCs) were subjected to immunofluorescence staining. A1 – D1 : DAPI staining. A2 – D2 : Immunofluorescence staining of mesothelin (green color). A3 – D3 : Immunofluorescence staining of NOX4 (red color). A4 – D4 : Overlays of immunofluorescence staining and DAPI staining, yellow areas represent colocalization of mesothelin and NOX4. DAPI, 4′,6-Diamidino-2-phenylindole dihydrochloride.

    Journal: Journal of Clinical Medicine

    Article Title: Inhibition of NADPH Oxidase 4 (NOX4) Signaling Attenuates Tuberculous Pleural Fibrosis

    doi: 10.3390/jcm8010116

    Figure Lengend Snippet: Activation of NOX4 and mesothelin deposition in a BCG-induced pleurisy mouse model. NOX4 and mesothelin (markers for PMCs) were subjected to immunofluorescence staining. A1 – D1 : DAPI staining. A2 – D2 : Immunofluorescence staining of mesothelin (green color). A3 – D3 : Immunofluorescence staining of NOX4 (red color). A4 – D4 : Overlays of immunofluorescence staining and DAPI staining, yellow areas represent colocalization of mesothelin and NOX4. DAPI, 4′,6-Diamidino-2-phenylindole dihydrochloride.

    Article Snippet: In brief, the sections were incubated with antibody to NOX4 (1:100 dilution, Santa Cruz Biotechnology, Santa Cruz, CA, USA) and antibody to mesothelin (1:100 dilutiob, Santa Cruz Biotechnology, Santa Cruz, CA, USA) overnight at 4 °C.

    Techniques: Activation Assay, Immunofluorescence, Staining

    CD26 high cells are cytotoxic and polyfunctional in vitro when engineered with a chimeric antigen receptor. A ) Transduction method. αCD3/ICOS-stimulated CD4 + T cell subsets were genetically engineered with a 1 st generation mesothelin-specific CAR. Cells were expanded for 6 days and analyzed by flow cytometry for CAR expression prior to use. B ) Percentage of K562-meso cells that were lysed by effector CD4 + T cell subsets. C ) Cytokine secretion determined by ELISA. Representative of 3 experiments.

    Journal: bioRxiv

    Article Title: Identification of human CD4+ T cell populations with distinct antitumor activity

    doi: 10.1101/2019.12.31.891317

    Figure Lengend Snippet: CD26 high cells are cytotoxic and polyfunctional in vitro when engineered with a chimeric antigen receptor. A ) Transduction method. αCD3/ICOS-stimulated CD4 + T cell subsets were genetically engineered with a 1 st generation mesothelin-specific CAR. Cells were expanded for 6 days and analyzed by flow cytometry for CAR expression prior to use. B ) Percentage of K562-meso cells that were lysed by effector CD4 + T cell subsets. C ) Cytokine secretion determined by ELISA. Representative of 3 experiments.

    Article Snippet: K562-meso cells were tested for mycoplasma (MycoAlert, Lonza) and mesothelin (R & D Systems, FAB32652) expression during expansion.

    Techniques: In Vitro, Transduction, Flow Cytometry, Expressing, Enzyme-linked Immunosorbent Assay

    Human CD26 high T cells ablate large human tumors and persist relative to other CD4 + T cell subsets. A ) ACT schematic. Th1 (CXCR3 + ), Th2 (CCR4 + ), Th17 (CCR4 + /CCR6 + ), CD26 high or bulk CD4 + cells were sorted from normal donor PBL and expanded with αCD3/ICOS bead at a 1 bead:10 T cell ratio. Cells were transduced with a 1 st generation mesothelin-specific CD3ζ CAR and expanded with IL-2. NSG mice bearing mesothelioma were treated with 4×10 6 transduced, sorted CD4 + cells + 4×10 6 transduced CD8 + cells and 50,000 IU IL-2 was given to each mouse daily for 3 days. B ) Single tumor curves overlaid with average curve (red) and C ) average tumor curves of 6-9 mice/group. All groups were significantly different from NT, P

    Journal: bioRxiv

    Article Title: Identification of human CD4+ T cell populations with distinct antitumor activity

    doi: 10.1101/2019.12.31.891317

    Figure Lengend Snippet: Human CD26 high T cells ablate large human tumors and persist relative to other CD4 + T cell subsets. A ) ACT schematic. Th1 (CXCR3 + ), Th2 (CCR4 + ), Th17 (CCR4 + /CCR6 + ), CD26 high or bulk CD4 + cells were sorted from normal donor PBL and expanded with αCD3/ICOS bead at a 1 bead:10 T cell ratio. Cells were transduced with a 1 st generation mesothelin-specific CD3ζ CAR and expanded with IL-2. NSG mice bearing mesothelioma were treated with 4×10 6 transduced, sorted CD4 + cells + 4×10 6 transduced CD8 + cells and 50,000 IU IL-2 was given to each mouse daily for 3 days. B ) Single tumor curves overlaid with average curve (red) and C ) average tumor curves of 6-9 mice/group. All groups were significantly different from NT, P

    Article Snippet: K562-meso cells were tested for mycoplasma (MycoAlert, Lonza) and mesothelin (R & D Systems, FAB32652) expression during expansion.

    Techniques: Transduction, Mouse Assay

    KLF8 alone is sufficient to induce T80 cells to form subcutaneous tumors resulting shortened life. A , Representative subcutaneous tumors formed by the indicated cell lines. Mock and SKOV3-ip1 were used as negative and positive controls, respectively. Tumorigenesis experiments were carried out as described in Materials and Methods. Photos were taken on 30 days (for SKOV3-ip1) or 105 days after injection. B , Tumor formation rates Tumor volumes were recorded at the indicated time points (see tumor incidence in Supplemental Table 1 ). C , T80/KLF8 tumor formation shortened mouse survival time. Survival distribution was presented by Kaplan-Meier curve. D , The T80/KLF8 tumors are highly similar to tumors of ovarian cancer patients. H E and IHC staining of the tumors were performed as described in Materials and Methods for expression of the KLF8 and human ovarian cancer marker proteins. a, H E (100X); b, KLF8 (100X); c, pan-cytokeratins (100X); d, CA 125 (100X); e, mesothelin (100X); f, HE4 (100X). Inlets, 400X

    Journal: Oncogene

    Article Title: Transformation of human ovarian surface epithelial cells by Kr?ppel-like factor 8

    doi: 10.1038/onc.2012.545

    Figure Lengend Snippet: KLF8 alone is sufficient to induce T80 cells to form subcutaneous tumors resulting shortened life. A , Representative subcutaneous tumors formed by the indicated cell lines. Mock and SKOV3-ip1 were used as negative and positive controls, respectively. Tumorigenesis experiments were carried out as described in Materials and Methods. Photos were taken on 30 days (for SKOV3-ip1) or 105 days after injection. B , Tumor formation rates Tumor volumes were recorded at the indicated time points (see tumor incidence in Supplemental Table 1 ). C , T80/KLF8 tumor formation shortened mouse survival time. Survival distribution was presented by Kaplan-Meier curve. D , The T80/KLF8 tumors are highly similar to tumors of ovarian cancer patients. H E and IHC staining of the tumors were performed as described in Materials and Methods for expression of the KLF8 and human ovarian cancer marker proteins. a, H E (100X); b, KLF8 (100X); c, pan-cytokeratins (100X); d, CA 125 (100X); e, mesothelin (100X); f, HE4 (100X). Inlets, 400X

    Article Snippet: The antibodies used in IHC include anti-KLF8 , anti-human CA125 (Clone M11, Dako), anti-human cytokeratin (Clones AE1/AE3, Dako), anti-cytokeratin 8 & 18 (Clone Zym5.2, Invitrogen), anti-HE4 (Covance) and anti-mesothelin (Novocastra).

    Techniques: Injection, Immunohistochemistry, Staining, Expressing, Marker

    KLF8 alone is sufficient to induce to induce T80 cells to form orthotopic ovarian tumors. A, Representative ovarian tumors formed by KLF8 expressing T80 cells. Mock and SKOV3-ip1 were used as negative and positive controls, respectively. 5 x 10 5 cells per cell lines were injected into the ovarian bursa. Photos of representative mice (top) and tumors (bottom) were taken 60 days or (SKOV3-ip1) 90 days (mock and KLF8) after injection. B , Tumor formation rate. The ovarian tumor volume recorded at the time of euthanasia is presented by box-plot (see tumor incidence in Supplemental Table 1 ). C . The T80/KLF8 ovarian tumors are highly similar to tumors of ovarian cancer patients. . H E and IHC staining of the tumors were performed as described in Materials and Methods for expression of the KLF8 and human ovarian cancer marker proteins. a, H E (100X); b, KLF8 (100X); c, pan-cytokeratins (100X); d, CA 125 (100X); e, mesothelin (100X); f, HE4 (100X). Inlets, 400X.

    Journal: Oncogene

    Article Title: Transformation of human ovarian surface epithelial cells by Kr?ppel-like factor 8

    doi: 10.1038/onc.2012.545

    Figure Lengend Snippet: KLF8 alone is sufficient to induce to induce T80 cells to form orthotopic ovarian tumors. A, Representative ovarian tumors formed by KLF8 expressing T80 cells. Mock and SKOV3-ip1 were used as negative and positive controls, respectively. 5 x 10 5 cells per cell lines were injected into the ovarian bursa. Photos of representative mice (top) and tumors (bottom) were taken 60 days or (SKOV3-ip1) 90 days (mock and KLF8) after injection. B , Tumor formation rate. The ovarian tumor volume recorded at the time of euthanasia is presented by box-plot (see tumor incidence in Supplemental Table 1 ). C . The T80/KLF8 ovarian tumors are highly similar to tumors of ovarian cancer patients. . H E and IHC staining of the tumors were performed as described in Materials and Methods for expression of the KLF8 and human ovarian cancer marker proteins. a, H E (100X); b, KLF8 (100X); c, pan-cytokeratins (100X); d, CA 125 (100X); e, mesothelin (100X); f, HE4 (100X). Inlets, 400X.

    Article Snippet: The antibodies used in IHC include anti-KLF8 , anti-human CA125 (Clone M11, Dako), anti-human cytokeratin (Clones AE1/AE3, Dako), anti-cytokeratin 8 & 18 (Clone Zym5.2, Invitrogen), anti-HE4 (Covance) and anti-mesothelin (Novocastra).

    Techniques: Expressing, Injection, Mouse Assay, Immunohistochemistry, Staining, Marker

    Scatterplot of calretinin versus mesothelin. The plot shows marker concentrations of MM cases and controls from Australia (group 2) and Germany (group 3)

    Journal: BMC Cancer

    Article Title: Calretinin as a blood-based biomarker for mesothelioma

    doi: 10.1186/s12885-017-3375-5

    Figure Lengend Snippet: Scatterplot of calretinin versus mesothelin. The plot shows marker concentrations of MM cases and controls from Australia (group 2) and Germany (group 3)

    Article Snippet: Determination of mesothelin For the determination of mesothelin in serum and plasma samples, a commercially available ELISA kit (MESOMARK) by Fujirebio Diagnostics, Inc. (Malvern, PA, USA) was used according to the manufacturer’s instructions as described before [ , ].

    Techniques: Marker

    Scatterplot of marker concentrations versus age. a Concentrations of calretinin [ng/mL] were plotted against age [years] of MM cases and controls. b Concentrations of mesothelin [nmol/L] were plotted against age [years] of MM cases and controls. The plots are based on pooled data from group 2 and 3

    Journal: BMC Cancer

    Article Title: Calretinin as a blood-based biomarker for mesothelioma

    doi: 10.1186/s12885-017-3375-5

    Figure Lengend Snippet: Scatterplot of marker concentrations versus age. a Concentrations of calretinin [ng/mL] were plotted against age [years] of MM cases and controls. b Concentrations of mesothelin [nmol/L] were plotted against age [years] of MM cases and controls. The plots are based on pooled data from group 2 and 3

    Article Snippet: Determination of mesothelin For the determination of mesothelin in serum and plasma samples, a commercially available ELISA kit (MESOMARK) by Fujirebio Diagnostics, Inc. (Malvern, PA, USA) was used according to the manufacturer’s instructions as described before [ , ].

    Techniques: Marker

    Marker concentrations in MM subtypes. a Calretinin [ng/mL] in controls and MM cases by subtype. b Mesothelin [nmol/L] in controls and MM cases by subtype. All cases and controls were from Australia (group 1). Individual p -values relate to the comparison between each subtype and the controls. P -values for calretinin were obtained from two-sided Peto-Prentice test and for mesothelin from two-sided Wilcoxon rank-sum test

    Journal: BMC Cancer

    Article Title: Calretinin as a blood-based biomarker for mesothelioma

    doi: 10.1186/s12885-017-3375-5

    Figure Lengend Snippet: Marker concentrations in MM subtypes. a Calretinin [ng/mL] in controls and MM cases by subtype. b Mesothelin [nmol/L] in controls and MM cases by subtype. All cases and controls were from Australia (group 1). Individual p -values relate to the comparison between each subtype and the controls. P -values for calretinin were obtained from two-sided Peto-Prentice test and for mesothelin from two-sided Wilcoxon rank-sum test

    Article Snippet: Determination of mesothelin For the determination of mesothelin in serum and plasma samples, a commercially available ELISA kit (MESOMARK) by Fujirebio Diagnostics, Inc. (Malvern, PA, USA) was used according to the manufacturer’s instructions as described before [ , ].

    Techniques: Marker

    Comparison of marker concentrations in samples from Australia and Germany. a Calretinin [ng/mL] in MM cases and controls from Australia (group 1 and 2) and Germany (group 3). The corresponding p -values (group 2 vs. group 3) are: p = 0.8210 for MM cases and p = 0.0773 for controls. b Mesothelin [nmol/L] in MM cases and controls from Australia (group 1 and 2) and Germany (group 3). The corresponding p -values (group 2 vs. group 3) are: p = 0.0012 for MM cases and p = 0.1422 for controls. P -values for calretinin were obtained from two-sided Peto-Prentice test and for mesothelin from two-sided Wilcoxon rank-sum test. For better comparison, for group 1 sarcomatoid MM were excluded

    Journal: BMC Cancer

    Article Title: Calretinin as a blood-based biomarker for mesothelioma

    doi: 10.1186/s12885-017-3375-5

    Figure Lengend Snippet: Comparison of marker concentrations in samples from Australia and Germany. a Calretinin [ng/mL] in MM cases and controls from Australia (group 1 and 2) and Germany (group 3). The corresponding p -values (group 2 vs. group 3) are: p = 0.8210 for MM cases and p = 0.0773 for controls. b Mesothelin [nmol/L] in MM cases and controls from Australia (group 1 and 2) and Germany (group 3). The corresponding p -values (group 2 vs. group 3) are: p = 0.0012 for MM cases and p = 0.1422 for controls. P -values for calretinin were obtained from two-sided Peto-Prentice test and for mesothelin from two-sided Wilcoxon rank-sum test. For better comparison, for group 1 sarcomatoid MM were excluded

    Article Snippet: Determination of mesothelin For the determination of mesothelin in serum and plasma samples, a commercially available ELISA kit (MESOMARK) by Fujirebio Diagnostics, Inc. (Malvern, PA, USA) was used according to the manufacturer’s instructions as described before [ , ].

    Techniques: Marker

    ROC analyses of calretinin and mesothelin with pooled data from Australia and Germany. a Nonparametric (AUC = 0.86, 95% CI = 0.82–0.91) and bi-lognormal (AUC = 0.90, 95% CI = 0.86–0.94) ROC curves for calretinin. b Nonparametric (AUC = 0.89, 95% CI = 0.85–0.93) and bi-Weibull (AUC = 0.91, 95% CI = 0.89–0.94) ROC curves for mesothelin. All ROC curves are based on pooled data from group 2 and 3

    Journal: BMC Cancer

    Article Title: Calretinin as a blood-based biomarker for mesothelioma

    doi: 10.1186/s12885-017-3375-5

    Figure Lengend Snippet: ROC analyses of calretinin and mesothelin with pooled data from Australia and Germany. a Nonparametric (AUC = 0.86, 95% CI = 0.82–0.91) and bi-lognormal (AUC = 0.90, 95% CI = 0.86–0.94) ROC curves for calretinin. b Nonparametric (AUC = 0.89, 95% CI = 0.85–0.93) and bi-Weibull (AUC = 0.91, 95% CI = 0.89–0.94) ROC curves for mesothelin. All ROC curves are based on pooled data from group 2 and 3

    Article Snippet: Determination of mesothelin For the determination of mesothelin in serum and plasma samples, a commercially available ELISA kit (MESOMARK) by Fujirebio Diagnostics, Inc. (Malvern, PA, USA) was used according to the manufacturer’s instructions as described before [ , ].

    Techniques:

    ROC analyses of calretinin and mesothelin in samples from Australia and Germany. a Nonparametric (AUC = 0.90, 95% CI = 0.85–0.95) and bi-lognormal (AUC = 0.95, 95% CI = 0.92–0.98) ROC curves for calretinin in Australian samples (group 2). b Nonparametric (AUC = 0.83, 95% CI = 0.74–0.92) and bi-lognormal (AUC = 0.87, 95% CI = 0.79–0.95) ROC curves for calretinin in German samples (group 3). c Nonparametric (AUC = 0.91, 95% CI = 0.87–0.96) and bi-Weibull (AUC = 0.93, 95% CI = 0.90–0.96) ROC curve for mesothelin in Australian samples (group 2). d Nonparametric (AUC = 0.84, 95% CI = 0.76–0.93) and bi-Weibull (AUC = 0.85, 95% CI = 0.81–0.89) ROC curve for mesothelin in German samples (group 3)

    Journal: BMC Cancer

    Article Title: Calretinin as a blood-based biomarker for mesothelioma

    doi: 10.1186/s12885-017-3375-5

    Figure Lengend Snippet: ROC analyses of calretinin and mesothelin in samples from Australia and Germany. a Nonparametric (AUC = 0.90, 95% CI = 0.85–0.95) and bi-lognormal (AUC = 0.95, 95% CI = 0.92–0.98) ROC curves for calretinin in Australian samples (group 2). b Nonparametric (AUC = 0.83, 95% CI = 0.74–0.92) and bi-lognormal (AUC = 0.87, 95% CI = 0.79–0.95) ROC curves for calretinin in German samples (group 3). c Nonparametric (AUC = 0.91, 95% CI = 0.87–0.96) and bi-Weibull (AUC = 0.93, 95% CI = 0.90–0.96) ROC curve for mesothelin in Australian samples (group 2). d Nonparametric (AUC = 0.84, 95% CI = 0.76–0.93) and bi-Weibull (AUC = 0.85, 95% CI = 0.81–0.89) ROC curve for mesothelin in German samples (group 3)

    Article Snippet: Determination of mesothelin For the determination of mesothelin in serum and plasma samples, a commercially available ELISA kit (MESOMARK) by Fujirebio Diagnostics, Inc. (Malvern, PA, USA) was used according to the manufacturer’s instructions as described before [ , ].

    Techniques:

    Immunohistochemical staining for mesothelin of HPAC tumor material from mice injected with 100 μg AMA-800CW (tumor harvested 144 h after injection) A. A representative brightfield image (400x) of a tumor with an IHC staining for mesothelin. B. Fluorescence microscopy at 800 nm for IRDye 800CW labelled to AMA; C. at 460 nm for DAPI staining, D. and overlay of fluorescent images.

    Journal: Oncotarget

    Article Title: Imaging the distribution of an antibody-drug conjugate constituent targeting mesothelin with 89Zr and IRDye 800CW in mice bearing human pancreatic tumor xenografts

    doi:

    Figure Lengend Snippet: Immunohistochemical staining for mesothelin of HPAC tumor material from mice injected with 100 μg AMA-800CW (tumor harvested 144 h after injection) A. A representative brightfield image (400x) of a tumor with an IHC staining for mesothelin. B. Fluorescence microscopy at 800 nm for IRDye 800CW labelled to AMA; C. at 460 nm for DAPI staining, D. and overlay of fluorescent images.

    Article Snippet: The extracellular domain (ECD) of mesothelin (provided by Genentech) was used to coat a 96-well plate (10 μg/mL, Nunc MaxiSorp, Thermo Fisher Scientific).

    Techniques: Immunohistochemistry, Staining, Mouse Assay, Injection, Fluorescence, Microscopy

    ELISA assay of binding affinity for mesothelin extra cellular domain with AMA conjugated to chelator, ratio 1:1.3 (yellow) and ratio 1:3.5 (red) compared to control (AMA, black) N = 3 for each ratio. X-axis depicts the amount of antibody added in nmol/mL; the Y-axis represents the optical density of the fluorescent signal at 450 nm.

    Journal: Oncotarget

    Article Title: Imaging the distribution of an antibody-drug conjugate constituent targeting mesothelin with 89Zr and IRDye 800CW in mice bearing human pancreatic tumor xenografts

    doi:

    Figure Lengend Snippet: ELISA assay of binding affinity for mesothelin extra cellular domain with AMA conjugated to chelator, ratio 1:1.3 (yellow) and ratio 1:3.5 (red) compared to control (AMA, black) N = 3 for each ratio. X-axis depicts the amount of antibody added in nmol/mL; the Y-axis represents the optical density of the fluorescent signal at 450 nm.

    Article Snippet: The extracellular domain (ECD) of mesothelin (provided by Genentech) was used to coat a 96-well plate (10 μg/mL, Nunc MaxiSorp, Thermo Fisher Scientific).

    Techniques: Enzyme-linked Immunosorbent Assay, Binding Assay

    Whole-blood IFN-γ responses to peptides spanning the MPF and mature mesothelin components 42 non-overlapping 15-mer peptides spanning the entire mesothelin molecules were exposed to peripheral blood of GBM patients over a seven-day period. Supernatants were then harvested for IFN-γ detection by ELISA. Absolute IFN-γ concentrations (pg/ml) produced by each patient for a single peptide A. as well as the average value per peptide (total IFN-γ/no. of patients) normalised to the sum of IFN-γ production for the entire peptide pool in percentage B. are shown. Immune hotspots within the mesothelin component, defined by peptide-specific IFN-γ production, identify several peptides which may represent viable targets to expand T-cells in host-directed therapies.

    Journal: Oncotarget

    Article Title: Mesothelin as a novel biomarker and immunotherapeutic target in human glioblastoma

    doi: 10.18632/oncotarget.20303

    Figure Lengend Snippet: Whole-blood IFN-γ responses to peptides spanning the MPF and mature mesothelin components 42 non-overlapping 15-mer peptides spanning the entire mesothelin molecules were exposed to peripheral blood of GBM patients over a seven-day period. Supernatants were then harvested for IFN-γ detection by ELISA. Absolute IFN-γ concentrations (pg/ml) produced by each patient for a single peptide A. as well as the average value per peptide (total IFN-γ/no. of patients) normalised to the sum of IFN-γ production for the entire peptide pool in percentage B. are shown. Immune hotspots within the mesothelin component, defined by peptide-specific IFN-γ production, identify several peptides which may represent viable targets to expand T-cells in host-directed therapies.

    Article Snippet: The mesothelin peptide pool is a customised product comprising 42 × 15-mer peptides without overlap covering the entire length of the mesothelin protein.

    Techniques: Enzyme-linked Immunosorbent Assay, Produced

    Recognition of mesothelin peptides by GBM TIL TIL isolated from GBM tumor tissue were cultured in vitro with IL-2/IL-15/IL-21 and exposed to mesothelin peptides over a 6-hour period. Cells were then stained with CD3, CD4, CD8, IFN-γ and TNF-α to visualise intracellular cytokine production by mesothelin-specific T-cells. Shown are frequencies (%) of responding cells based on CD3+ TIL of one representative patient. Cell frequency of more than 0.2% is a considered a legitimate response.

    Journal: Oncotarget

    Article Title: Mesothelin as a novel biomarker and immunotherapeutic target in human glioblastoma

    doi: 10.18632/oncotarget.20303

    Figure Lengend Snippet: Recognition of mesothelin peptides by GBM TIL TIL isolated from GBM tumor tissue were cultured in vitro with IL-2/IL-15/IL-21 and exposed to mesothelin peptides over a 6-hour period. Cells were then stained with CD3, CD4, CD8, IFN-γ and TNF-α to visualise intracellular cytokine production by mesothelin-specific T-cells. Shown are frequencies (%) of responding cells based on CD3+ TIL of one representative patient. Cell frequency of more than 0.2% is a considered a legitimate response.

    Article Snippet: The mesothelin peptide pool is a customised product comprising 42 × 15-mer peptides without overlap covering the entire length of the mesothelin protein.

    Techniques: Isolation, Cell Culture, In Vitro, Staining

    Whole-blood IFN-γ responses of glioma patients to the mesothelin precursor protein, MPF and the mature mesothelin component with or without cytokine conditioning Whole-blood obtained from patients with glioma (GBM, A ; astrocytoma, B ; OD, C ; metastasis, D ) were cultured with mesothelin or its derivatives in the absence of cytokine conditioning, with IL-2/IL-7 or IL-2/IL-15/IL-21 conditioning over seven days. Supernatants were then harvested for IFN-γ detection by ELISA. Shown are dot plots representing responses of individual patients. Mann Whitney test of medians was performed to gauge statistical significance. * p

    Journal: Oncotarget

    Article Title: Mesothelin as a novel biomarker and immunotherapeutic target in human glioblastoma

    doi: 10.18632/oncotarget.20303

    Figure Lengend Snippet: Whole-blood IFN-γ responses of glioma patients to the mesothelin precursor protein, MPF and the mature mesothelin component with or without cytokine conditioning Whole-blood obtained from patients with glioma (GBM, A ; astrocytoma, B ; OD, C ; metastasis, D ) were cultured with mesothelin or its derivatives in the absence of cytokine conditioning, with IL-2/IL-7 or IL-2/IL-15/IL-21 conditioning over seven days. Supernatants were then harvested for IFN-γ detection by ELISA. Shown are dot plots representing responses of individual patients. Mann Whitney test of medians was performed to gauge statistical significance. * p

    Article Snippet: The mesothelin peptide pool is a customised product comprising 42 × 15-mer peptides without overlap covering the entire length of the mesothelin protein.

    Techniques: Cell Culture, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY

    Whole-blood IFN-γ responses to the mesothelin precursor protein and its derivatives with or without cytokine conditioning based on glioma grading The data presented in Figure 1 was reanalysed based on WHO tumour grading, grade I being the least malignant and grade IV being the most aggressive (see ‘Results’). Shown are dot plots representing responses of individual patients. Mann Whitney test of medians was performed to gauge statistical significance. * p

    Journal: Oncotarget

    Article Title: Mesothelin as a novel biomarker and immunotherapeutic target in human glioblastoma

    doi: 10.18632/oncotarget.20303

    Figure Lengend Snippet: Whole-blood IFN-γ responses to the mesothelin precursor protein and its derivatives with or without cytokine conditioning based on glioma grading The data presented in Figure 1 was reanalysed based on WHO tumour grading, grade I being the least malignant and grade IV being the most aggressive (see ‘Results’). Shown are dot plots representing responses of individual patients. Mann Whitney test of medians was performed to gauge statistical significance. * p

    Article Snippet: The mesothelin peptide pool is a customised product comprising 42 × 15-mer peptides without overlap covering the entire length of the mesothelin protein.

    Techniques: MANN-WHITNEY

    GBM peripheral blood T-cell proliferation in response to mesothelin stimulation with or without cytokine conditioning Immune cells remaining in the culture plate at the end of the whole blood assay were purified, processed and stained with CD3, CD4 and CD8 antibodies for flow cytometric analysis Shown are dot plots representing responses of individual patients. Mann Whitney test of medians was performed to gauge statistical significance. * p

    Journal: Oncotarget

    Article Title: Mesothelin as a novel biomarker and immunotherapeutic target in human glioblastoma

    doi: 10.18632/oncotarget.20303

    Figure Lengend Snippet: GBM peripheral blood T-cell proliferation in response to mesothelin stimulation with or without cytokine conditioning Immune cells remaining in the culture plate at the end of the whole blood assay were purified, processed and stained with CD3, CD4 and CD8 antibodies for flow cytometric analysis Shown are dot plots representing responses of individual patients. Mann Whitney test of medians was performed to gauge statistical significance. * p

    Article Snippet: The mesothelin peptide pool is a customised product comprising 42 × 15-mer peptides without overlap covering the entire length of the mesothelin protein.

    Techniques: Whole Blood Assay, Purification, Staining, Flow Cytometry, MANN-WHITNEY

    Detection of mesothelin-specific circulating IgG as well as shed mesothelin protein in plasma of GBM patients An indirect ELISA method was used for quantifying mesothelin-specific IgG titres in plasma, while a commercially available ELISA kit was used for measuring mesothelin protein. IgG titres and mesothelin levels are expressed as ng/ml of plasma. Mann Whitney test of medians was performed to gauge statistical significance. * p

    Journal: Oncotarget

    Article Title: Mesothelin as a novel biomarker and immunotherapeutic target in human glioblastoma

    doi: 10.18632/oncotarget.20303

    Figure Lengend Snippet: Detection of mesothelin-specific circulating IgG as well as shed mesothelin protein in plasma of GBM patients An indirect ELISA method was used for quantifying mesothelin-specific IgG titres in plasma, while a commercially available ELISA kit was used for measuring mesothelin protein. IgG titres and mesothelin levels are expressed as ng/ml of plasma. Mann Whitney test of medians was performed to gauge statistical significance. * p

    Article Snippet: The mesothelin peptide pool is a customised product comprising 42 × 15-mer peptides without overlap covering the entire length of the mesothelin protein.

    Techniques: Indirect ELISA, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY

    Immunohistological confirmation of mesothelin overexpression in GBM tissue Paraffin-embedded tissue sections were stained with rat anti-human mesothelin primary antibody. Detection was performed using a goat anti-rat IgG secondary antibody labelled with Alexa Fluor 488-conjugated polyclonal goat anti-rat secondary antibody. The primary antibody was omitted in the negative control while the pancreatic cancer tumour cell line PaTu was used as positive control ( Supplementary Figure 1 ). Shown are representative photographs for GBM tissue from one patient.

    Journal: Oncotarget

    Article Title: Mesothelin as a novel biomarker and immunotherapeutic target in human glioblastoma

    doi: 10.18632/oncotarget.20303

    Figure Lengend Snippet: Immunohistological confirmation of mesothelin overexpression in GBM tissue Paraffin-embedded tissue sections were stained with rat anti-human mesothelin primary antibody. Detection was performed using a goat anti-rat IgG secondary antibody labelled with Alexa Fluor 488-conjugated polyclonal goat anti-rat secondary antibody. The primary antibody was omitted in the negative control while the pancreatic cancer tumour cell line PaTu was used as positive control ( Supplementary Figure 1 ). Shown are representative photographs for GBM tissue from one patient.

    Article Snippet: The mesothelin peptide pool is a customised product comprising 42 × 15-mer peptides without overlap covering the entire length of the mesothelin protein.

    Techniques: Over Expression, Staining, Negative Control, Positive Control

    Plasma endpoint biomarkers. Rats were injected with 100 μL serum free media or serum free media containing 5 × 10 5 parental or chemo-resistant II-45 cells directly into the pleural cavity. At ethical endpoint, rats were euthanized and plasma samples were analysed for cytokine levels relative to rats with parental II-45 mesothelioma. (a) osteopontin levels; (b) mesothelin levels. Normal control rats, Control; parental mesothelioma cells, II-45; cisplatin resistant II-45 cells, CisR; pemetrexed resistant II-45 cells, PemR; combination (cisplatin + pemetrexed) resistant II-45 cells, ComboR; gemcitabine resistant II-45 cells, GemR; vinorelbine resistant II-45 cells, VLBR. P-values were calculated using a one-way Anova test with a value of less than 0.05 indicating significance. * p

    Journal: Scientific Reports

    Article Title: Establishing a panel of chemo-resistant mesothelioma models for investigating chemo-resistance and identifying new treatments for mesothelioma

    doi: 10.1038/srep06152

    Figure Lengend Snippet: Plasma endpoint biomarkers. Rats were injected with 100 μL serum free media or serum free media containing 5 × 10 5 parental or chemo-resistant II-45 cells directly into the pleural cavity. At ethical endpoint, rats were euthanized and plasma samples were analysed for cytokine levels relative to rats with parental II-45 mesothelioma. (a) osteopontin levels; (b) mesothelin levels. Normal control rats, Control; parental mesothelioma cells, II-45; cisplatin resistant II-45 cells, CisR; pemetrexed resistant II-45 cells, PemR; combination (cisplatin + pemetrexed) resistant II-45 cells, ComboR; gemcitabine resistant II-45 cells, GemR; vinorelbine resistant II-45 cells, VLBR. P-values were calculated using a one-way Anova test with a value of less than 0.05 indicating significance. * p

    Article Snippet: Plasma levels for mesothelin and osteopontin were determined using the rat N-ERC/mesothelin (Immuno-Biological Laboratories, Gunma, Japan) and the rat osteopontin (USCN Life Sciences, Houston, Texas, USA) ELISA kits respectively.

    Techniques: Injection