anti human ifn γ  (Thermo Fisher)


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  • 99
    Name:
    IFN gamma Antibody 11H20L4
    Description:
    IFN gamma Monoclonal Antibody for Western Blot
    Catalog Number:
    701263
    Price:
    None
    Category:
    Antibodies Secondary Detection Reagents
    Applications:
    Cell Analysis|ELISA|ELISAs for Cytokine, Chemokine & Growth Factors|Protein Assays and Analysis|Protein Biology|Ready-To-Use Immunoassay|Western Blot Detection|Western Blotting
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    Structured Review

    Thermo Fisher anti human ifn γ
    PSGR-derived peptide-induced T cell responses were CD8 + T cell dependent and restricted by HLA-I. PSGR-derived peptide-specific T cells were co-cultured with T2 cells pulsed with or without a given peptide in the presence of GolgiStop in a 48-well plate for 4 hrs at 37°C. Cells were stained with anti-CD8 and <t>anti-IFN-γ,</t> then analyzed on a FACScalibur machine (A). PSGR-derived peptide-specific T cells were co-incubated with LNCaP cells alone in medium, or with LNCaP cells in the presence of either anti-HLA-I mAb (W6/32), HLA-II mAb or a control mAb (anti-CD19 mAb). After 4 hours of incubation, the cytotoxicity against LNCaP was determined by the LDH assay (B). Data from B are plotted as means ± SD. Results are representative of three independent experiments. * P
    IFN gamma Monoclonal Antibody for Western Blot
    https://www.bioz.com/result/anti human ifn γ/product/Thermo Fisher
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti human ifn γ - by Bioz Stars, 2021-05
    99/100 stars

    Images

    1) Product Images from "Identification of Prostate-Specific G-Protein Coupled Receptor as a Tumor Antigen Recognized by CD8+ T Cells for Cancer Immunotherapy"

    Article Title: Identification of Prostate-Specific G-Protein Coupled Receptor as a Tumor Antigen Recognized by CD8+ T Cells for Cancer Immunotherapy

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0045756

    PSGR-derived peptide-induced T cell responses were CD8 + T cell dependent and restricted by HLA-I. PSGR-derived peptide-specific T cells were co-cultured with T2 cells pulsed with or without a given peptide in the presence of GolgiStop in a 48-well plate for 4 hrs at 37°C. Cells were stained with anti-CD8 and anti-IFN-γ, then analyzed on a FACScalibur machine (A). PSGR-derived peptide-specific T cells were co-incubated with LNCaP cells alone in medium, or with LNCaP cells in the presence of either anti-HLA-I mAb (W6/32), HLA-II mAb or a control mAb (anti-CD19 mAb). After 4 hours of incubation, the cytotoxicity against LNCaP was determined by the LDH assay (B). Data from B are plotted as means ± SD. Results are representative of three independent experiments. * P
    Figure Legend Snippet: PSGR-derived peptide-induced T cell responses were CD8 + T cell dependent and restricted by HLA-I. PSGR-derived peptide-specific T cells were co-cultured with T2 cells pulsed with or without a given peptide in the presence of GolgiStop in a 48-well plate for 4 hrs at 37°C. Cells were stained with anti-CD8 and anti-IFN-γ, then analyzed on a FACScalibur machine (A). PSGR-derived peptide-specific T cells were co-incubated with LNCaP cells alone in medium, or with LNCaP cells in the presence of either anti-HLA-I mAb (W6/32), HLA-II mAb or a control mAb (anti-CD19 mAb). After 4 hours of incubation, the cytotoxicity against LNCaP was determined by the LDH assay (B). Data from B are plotted as means ± SD. Results are representative of three independent experiments. * P

    Techniques Used: Derivative Assay, Cell Culture, Staining, Incubation, Lactate Dehydrogenase Assay

    PSGR-derived peptides induced peptide-specific T cells. The recognition of T2 cells pre-loaded with titrated concentrations of peptides (0–20 µg/ml) by expanded PSGR peptide-specific T cells was tested by ELISA assay (A). The expanded PSGR3 T cells (B and E), PSGR4 T cells (C and F) and PSGR14 T cells (D and G) were respectively co-incubated with T2 cells (1×10 4 cells/well) alone in complete medium (CM), or with T2 cells pre-loaded with either a corresponding peptide (5 µg/mL) or a control peptide as a negative control. Cells were incubated for 18 −24 hours, the IFN-γ secretion in the supernatant was determined by ELISA assay (B, C and D). IFN-γ spot-forming cells (SFC) were enumerated by ELISPOT assay (E, F and G). Data are plotted as means ± SD. Results are representative of at least three independent experiments. * P
    Figure Legend Snippet: PSGR-derived peptides induced peptide-specific T cells. The recognition of T2 cells pre-loaded with titrated concentrations of peptides (0–20 µg/ml) by expanded PSGR peptide-specific T cells was tested by ELISA assay (A). The expanded PSGR3 T cells (B and E), PSGR4 T cells (C and F) and PSGR14 T cells (D and G) were respectively co-incubated with T2 cells (1×10 4 cells/well) alone in complete medium (CM), or with T2 cells pre-loaded with either a corresponding peptide (5 µg/mL) or a control peptide as a negative control. Cells were incubated for 18 −24 hours, the IFN-γ secretion in the supernatant was determined by ELISA assay (B, C and D). IFN-γ spot-forming cells (SFC) were enumerated by ELISPOT assay (E, F and G). Data are plotted as means ± SD. Results are representative of at least three independent experiments. * P

    Techniques Used: Derivative Assay, Enzyme-linked Immunosorbent Assay, Incubation, Negative Control, Enzyme-linked Immunospot

    2) Product Images from "Control of IFN-γ production and regulatory function by the inducible nuclear protein IκB-ζ in T cells"

    Article Title: Control of IFN-γ production and regulatory function by the inducible nuclear protein IκB-ζ in T cells

    Journal: Journal of Leukocyte Biology

    doi: 10.1189/jlb.2A0814-384R

    IκB-ζ-deficient T cells show high levels of IFN-γ expression in the presence of TGF-β.
    Figure Legend Snippet: IκB-ζ-deficient T cells show high levels of IFN-γ expression in the presence of TGF-β.

    Techniques Used: Expressing

    3) Product Images from "T Helper Cell Subsets Specific for Pseudomonas aeruginosa in Healthy Individuals and Patients with Cystic Fibrosis"

    Article Title: T Helper Cell Subsets Specific for Pseudomonas aeruginosa in Healthy Individuals and Patients with Cystic Fibrosis

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0090263

    Cytokine production by human memory CD4 + T cells in healthy controls and cystic fibrosis. Human memory CD4 + T cells were exposed to unstimulated DCs or DCs infected with the laboratory strains (PA103) or clinical Pseudomonas aeruginosa strains (Yorkhill). After 6 days of co-culture levels of ( A ) IL-17, ( B ) IL-22 and ( C ) IFN-γ were measured. Each point represents the mean of triplicate wells and is the result from one individual. The different bacterial strains used are: ♦, unstimulated; ○, PA103 ΔpcrV; •, PA103ΔUΔT; ▴, YH1; ▪, YH2; ×, YH5. The line indicates the median value. Differences between infection conditions ( A – C ) were evaluated by a Kruskal-Wallis test all of which had p values
    Figure Legend Snippet: Cytokine production by human memory CD4 + T cells in healthy controls and cystic fibrosis. Human memory CD4 + T cells were exposed to unstimulated DCs or DCs infected with the laboratory strains (PA103) or clinical Pseudomonas aeruginosa strains (Yorkhill). After 6 days of co-culture levels of ( A ) IL-17, ( B ) IL-22 and ( C ) IFN-γ were measured. Each point represents the mean of triplicate wells and is the result from one individual. The different bacterial strains used are: ♦, unstimulated; ○, PA103 ΔpcrV; •, PA103ΔUΔT; ▴, YH1; ▪, YH2; ×, YH5. The line indicates the median value. Differences between infection conditions ( A – C ) were evaluated by a Kruskal-Wallis test all of which had p values

    Techniques Used: Infection, Co-Culture Assay

    Tissue-homing characteristics of Pseudomonas aeruginosa-specific memory Th22 and Th17 cells. A–C Human memory CD4 + T cells were sorted into CCR6-depleted (CCR6 negative) and CCR6-enriched (CCR6 positive) populations before being exposed to dendritic cells (DCs) infected with Yorkhill 5 MOI30 ( A ), DCs infected with PA103 ΔUΔT MOI30 ( B ), or stimulated with anti-CD3 and anti-CD28 which acted as a positive control by polyclonal T cell stimulation ( C ). Plots show CD4 + IFN-γ negative T cells after 6-days of co-culture. D Memory CD4 + T cells were cultured with DCs infected with PA103 ΔpcrV MOI30. Pseudomonas-specific memory CD4 + T cells were analysed for expression of intracellular IL-22 or IL-17 as shown together with the skin-homing chemokine receptors CCR4, CCR10 and CCR9 as indicated. Numbers in each plot represent per cent cells in each quadrant. Results are representative of those seen in three independent individuals.
    Figure Legend Snippet: Tissue-homing characteristics of Pseudomonas aeruginosa-specific memory Th22 and Th17 cells. A–C Human memory CD4 + T cells were sorted into CCR6-depleted (CCR6 negative) and CCR6-enriched (CCR6 positive) populations before being exposed to dendritic cells (DCs) infected with Yorkhill 5 MOI30 ( A ), DCs infected with PA103 ΔUΔT MOI30 ( B ), or stimulated with anti-CD3 and anti-CD28 which acted as a positive control by polyclonal T cell stimulation ( C ). Plots show CD4 + IFN-γ negative T cells after 6-days of co-culture. D Memory CD4 + T cells were cultured with DCs infected with PA103 ΔpcrV MOI30. Pseudomonas-specific memory CD4 + T cells were analysed for expression of intracellular IL-22 or IL-17 as shown together with the skin-homing chemokine receptors CCR4, CCR10 and CCR9 as indicated. Numbers in each plot represent per cent cells in each quadrant. Results are representative of those seen in three independent individuals.

    Techniques Used: Infection, Positive Control, Cell Stimulation, Co-Culture Assay, Cell Culture, Expressing

    Cytokine production of human memory CD4 + T cells in response to P. aeruginosa . A–C Sorted human memory CD4 + T cells from healthy volunteers were co-cultured either with dendritic cells that had been infected with P. aeruginosa (DCs +) or with the supernatant from such infected DCs (DC −). Levels of IL-17A ( A ), IL-22 ( B ) and IFN-γ ( C ) were measured in supernatants after 6 days. DCs were infected at a MOI of 60 before culturing with T cells. Columns show the mean of triplicate determinations; error bars are ±1 standard error of mean. Results are representative of experiments in three separate individuals. D–F , secreted cytokine levels following DC infection at a variety of different MOIs. Each point represents a value from an individual healthy individual.
    Figure Legend Snippet: Cytokine production of human memory CD4 + T cells in response to P. aeruginosa . A–C Sorted human memory CD4 + T cells from healthy volunteers were co-cultured either with dendritic cells that had been infected with P. aeruginosa (DCs +) or with the supernatant from such infected DCs (DC −). Levels of IL-17A ( A ), IL-22 ( B ) and IFN-γ ( C ) were measured in supernatants after 6 days. DCs were infected at a MOI of 60 before culturing with T cells. Columns show the mean of triplicate determinations; error bars are ±1 standard error of mean. Results are representative of experiments in three separate individuals. D–F , secreted cytokine levels following DC infection at a variety of different MOIs. Each point represents a value from an individual healthy individual.

    Techniques Used: Cell Culture, Infection

    4) Product Images from "Contrasting roles for all-trans retinoic acid in TGF-?-mediated induction of Foxp3 and Il10 genes in developing regulatory T cells"

    Article Title: Contrasting roles for all-trans retinoic acid in TGF-?-mediated induction of Foxp3 and Il10 genes in developing regulatory T cells

    Journal: The Journal of Experimental Medicine

    doi: 10.1084/jem.20080950

    The combination of TGF-β– and CpG-activated BMDC induces IL-10–expressing CD4 T cells . (A) 10 5 D10 BMDC that were either left unactivated or preactivated with the indicated agonists were used in combination with 1 µg/ml OVA peptide to stimulate 5 × 10 5 naive CD4 T cells from 10BiT. Foxp3 gfp .OT-II mice, in the absence (top) or presence (bottom) of 5 ng/ml of recombinant human TGF-β1. On day 3, CD4 T cells were examined for expression of Thy1.1 and GFP. Numbers represent the percentage of CD4 T cells in the respective quadrants. (B) Naive CD4 T cells were stimulated as described in A with CpG-BMDC in the presence of TGF-β only or TGF-β plus anti–IFN-γ and anti–IL-4 (each at a final concentration of 10 µg/ml). On day 5, cells were restimulated for 5 h with PMA/ionomycin in the presence of brefeldin A, and viable CD4 T cells were analyzed for expression of Thy1.1, Foxp3, IL-17A, and IFN-γ. (C) Day-5 Thy1.1 + T cells (induced in the presence of CpG-BMDC and TGF-β) were MACS sorted and cultured for 24 h with Il10 −/− splenic DC in the absence (−) or presence (+) of 1 µg/ml OVA peptide. IL-10 protein in culture supernatants was quantitated by cytometric bead array. Mean cytokine concentrations ±SEM are shown. (ND, not detected). (D) 5 × 10 5 CFSE-labeled OT-II CD4 T cells were stimulated with 1.0 µg/ml OVA peptide and 10 5 IL-10–deficient splenic DC in the lower chamber of a transwell plate. In some cases, an equal number of DC or DC plus 5 × 10 5 day-5 Thy1.1 + T cells (differentiated as in C) were similarly activated in the upper chamber. Where indicated, anti–IL-10R antibody was added to the lower chamber at a final concentration of 10 µg/ml. After 4 d, cells were harvested from the bottom chamber and the CFSE profile of the CD4 T cells was analyzed by FACS. Numbers in A and B indicate the percentages of CD4 T cells in the given quadrants. All data are representative of at least two experiments.
    Figure Legend Snippet: The combination of TGF-β– and CpG-activated BMDC induces IL-10–expressing CD4 T cells . (A) 10 5 D10 BMDC that were either left unactivated or preactivated with the indicated agonists were used in combination with 1 µg/ml OVA peptide to stimulate 5 × 10 5 naive CD4 T cells from 10BiT. Foxp3 gfp .OT-II mice, in the absence (top) or presence (bottom) of 5 ng/ml of recombinant human TGF-β1. On day 3, CD4 T cells were examined for expression of Thy1.1 and GFP. Numbers represent the percentage of CD4 T cells in the respective quadrants. (B) Naive CD4 T cells were stimulated as described in A with CpG-BMDC in the presence of TGF-β only or TGF-β plus anti–IFN-γ and anti–IL-4 (each at a final concentration of 10 µg/ml). On day 5, cells were restimulated for 5 h with PMA/ionomycin in the presence of brefeldin A, and viable CD4 T cells were analyzed for expression of Thy1.1, Foxp3, IL-17A, and IFN-γ. (C) Day-5 Thy1.1 + T cells (induced in the presence of CpG-BMDC and TGF-β) were MACS sorted and cultured for 24 h with Il10 −/− splenic DC in the absence (−) or presence (+) of 1 µg/ml OVA peptide. IL-10 protein in culture supernatants was quantitated by cytometric bead array. Mean cytokine concentrations ±SEM are shown. (ND, not detected). (D) 5 × 10 5 CFSE-labeled OT-II CD4 T cells were stimulated with 1.0 µg/ml OVA peptide and 10 5 IL-10–deficient splenic DC in the lower chamber of a transwell plate. In some cases, an equal number of DC or DC plus 5 × 10 5 day-5 Thy1.1 + T cells (differentiated as in C) were similarly activated in the upper chamber. Where indicated, anti–IL-10R antibody was added to the lower chamber at a final concentration of 10 µg/ml. After 4 d, cells were harvested from the bottom chamber and the CFSE profile of the CD4 T cells was analyzed by FACS. Numbers in A and B indicate the percentages of CD4 T cells in the given quadrants. All data are representative of at least two experiments.

    Techniques Used: Expressing, Mouse Assay, Recombinant, Concentration Assay, Magnetic Cell Separation, Cell Culture, Labeling, FACS

    5) Product Images from "The Serine-threonine Kinase Inositol-requiring Enzyme 1α (IRE1α) Promotes IL-4 Production in T Helper Cells *"

    Article Title: The Serine-threonine Kinase Inositol-requiring Enzyme 1α (IRE1α) Promotes IL-4 Production in T Helper Cells *

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M113.493171

    IRE1α KO-activated CD4 + T cells produce less IL-4 than control in response to TCR stimulation. A–D , less KO Th0 cells make IL-4 than control cells. IL-4 and IFN-γ were measured by ICCS in cells activated for 5 days under Th0 conditions
    Figure Legend Snippet: IRE1α KO-activated CD4 + T cells produce less IL-4 than control in response to TCR stimulation. A–D , less KO Th0 cells make IL-4 than control cells. IL-4 and IFN-γ were measured by ICCS in cells activated for 5 days under Th0 conditions

    Techniques Used:

    Related Articles

    Enzyme-linked Immunospot:

    Article Title: Identification of Prostate-Specific G-Protein Coupled Receptor as a Tumor Antigen Recognized by CD8+ T Cells for Cancer Immunotherapy
    Article Snippet: IFN-γ ELISPOT Assay The IFN-γ ELISPOT assay was performed as previously described to quantify peptide-specific cytotoxic T-lymphocytes (CTLs) after in vitro expansion. .. Briefly, 96-well ELISPOT plates (Millipore; Bedford, MA, USA) were coated overnight at 4°C with 7.5 µg/mL anti-human IFN-γ (Pierce Biotechnology; Rockford, IL, USA). ..

    Blocking Assay:

    Article Title: Impaired CD4+ T cell proliferation and effector function correlates with repressive histone methylation events in a mouse model of severe sepsis
    Article Snippet: For in vitro skewing, culture medium was supplanted with the following recombinant cytokines when indicated: 10 U/ml IL-2 (Peprotech, Rocky Hill, NJ), 10 ng/ml IL-4 or IL-12 (R and D Systems, Minneapolis, MN). .. Additionally, the following blocking antibodies were used when indicated: α-IL-4, α-IL-12, and α-IFN-γ, all at 10 μg/ml (eBioscience, San Diego, CA). .. For analysis of in vitro proliferation, freshly isolated CD4+ CD62L- and CD4+ CD62L+ T cells from sham and CLP mice 14 days post-surgery were stimulated with plate-bound α-CD3 and soluble α-CD28 for 72 hours.

    Staining:

    Article Title: T Helper Cell Subsets Specific for Pseudomonas aeruginosa in Healthy Individuals and Patients with Cystic Fibrosis
    Article Snippet: Extracellular staining was with phycoerythrin-labelled anti-CD4 (RPA-T4; eBioscience), phycoerythrin-labelled anti-αβTCR (IP26; eBioscience), phycoerythrin-Cy7-labelled anti-CCR4 (IGI; BD Pharminogen), AlexFluor®647-labelled anti-CCR9 (BL/CCR9; BioLegend), phycoerythrin-labelled anti-CCR10 (FAB 3478P; R & D), CD86 and CD40. .. Intracellular staining was with eFluor®660-labelled anti-IL-22 (22URTI; eBioscience), phycoerythrin-labelled anti-IL-22 (BG/IL-22; Biolegend), PerCP-Cy5.5-labelled anti-IFN-γ (4S.B3; eBioscience), and AlexFluor®488 anti-IL-17A (eBioDEC17; eBioscience). ..

    Article Title: IRE1α-XBP1 controls T cell function in ovarian cancer by regulating mitochondrial activity
    Article Snippet: Flow cytometry-based analyses Flow cytometry was conducted using fluorochrome-conjugated antibodies purchased from BioLegend, unless stated otherwise. .. For staining of mouse cells we used: anti-CD45 (30-F11), anti-CD4 (RM4–5), anti-CD8 (53–6.7), anti-CD3 (145–2C11), anti-TCRβ (H57–597), anti-CD44 (IM7), anti-CD62L (MEL-14), anti-CD11b (M1/70), anti-CD11c (N418), anti-CD19 (6D5), anti-F4/80 (BM8), anti-Ly6C (HK1.4), anti-Ly6G (1A8), anti-I-A/I-E (M5/114.15.2), anti-IFN-γ (XMG1.2), anti-IL-17A (TC11–18H10.1), anti-FoxP3 (150D), anti-PD-1 (29F.1A12), anti-CTLA4 (UC10–4B9), anti-TIM3 (B8.2C12), TruStain fcX CD16/32 (93); anti-Perforin (17–9392-80, eBioscience). .. To stain human cells we utilized: anti-CD45 (HI30), anti-CD3 (HIT3), anti-CD4 (A161A1), anti-CD8 (HIT8a), anti-CD20 (2H7), anti-CD14 (63D3), anti-IFN-γ (4S.B3), anti-T-bet (4B10), TruStain FcX solution (422302); anti-XBP1s (Q3–695; BD Pharmingen); anti-RORγt (AFKJS-9, eBioscience).

    Functional Assay:

    Article Title: Contrasting roles for all-trans retinoic acid in TGF-?-mediated induction of Foxp3 and Il10 genes in developing regulatory T cells
    Article Snippet: All mice were bred and maintained and all animal experimentation was approved in accordance with guidelines and approval of the University of Alabama at Birmingham Institutional Animal Care and Use Committee. .. PE-conjugated anti-CD44 (IM7), anti-α4 β7 (LPAM), anti-CD90.1 (OX-7), anti-CD103 (M290), anti-IL-17 (TC11-18H10), PerCP (peridinin chlorophyll-a protein)-conjugated anti-CD90.1 (OX-7), and functional grade anti-IL-10R (1b1.3a) were purchased from BD; allophycocyanin-conjugated anti-CD4 (GK1.5), anti-IL-10 (JES5-16S3), anti–IFN-γ (XMG1.2), PE-conjugated and anti–IL-10 (JES5-16S3), PE-Cy7–conjugated anti-CD4 (GK1.5), affinity-purified anti-CCR9 (CW-1.2), and functional grade anti–IL-6 (MP5-20F3) were purchased from eBioscience. .. The following reagents were purchased from R & D Systems: recombinant human TGF-β1, IL-21 R/Fc chimera, WSSX-1 R/Fc chimera, and anti–IFN-α/βR1. at-RA was purchased from Sigma-Aldrich and the RA inhibitor LE540 was purchased from Wako Chemicals USA, Inc. MDP, LTA, poly (I:C), LPS, and CpG ODN were purchased from InvivoGen.

    Affinity Purification:

    Article Title: Contrasting roles for all-trans retinoic acid in TGF-?-mediated induction of Foxp3 and Il10 genes in developing regulatory T cells
    Article Snippet: All mice were bred and maintained and all animal experimentation was approved in accordance with guidelines and approval of the University of Alabama at Birmingham Institutional Animal Care and Use Committee. .. PE-conjugated anti-CD44 (IM7), anti-α4 β7 (LPAM), anti-CD90.1 (OX-7), anti-CD103 (M290), anti-IL-17 (TC11-18H10), PerCP (peridinin chlorophyll-a protein)-conjugated anti-CD90.1 (OX-7), and functional grade anti-IL-10R (1b1.3a) were purchased from BD; allophycocyanin-conjugated anti-CD4 (GK1.5), anti-IL-10 (JES5-16S3), anti–IFN-γ (XMG1.2), PE-conjugated and anti–IL-10 (JES5-16S3), PE-Cy7–conjugated anti-CD4 (GK1.5), affinity-purified anti-CCR9 (CW-1.2), and functional grade anti–IL-6 (MP5-20F3) were purchased from eBioscience. .. The following reagents were purchased from R & D Systems: recombinant human TGF-β1, IL-21 R/Fc chimera, WSSX-1 R/Fc chimera, and anti–IFN-α/βR1. at-RA was purchased from Sigma-Aldrich and the RA inhibitor LE540 was purchased from Wako Chemicals USA, Inc. MDP, LTA, poly (I:C), LPS, and CpG ODN were purchased from InvivoGen.

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  • 99
    Thermo Fisher anti human ifn γ
    PSGR-derived peptide-induced T cell responses were CD8 + T cell dependent and restricted by HLA-I. PSGR-derived peptide-specific T cells were co-cultured with T2 cells pulsed with or without a given peptide in the presence of GolgiStop in a 48-well plate for 4 hrs at 37°C. Cells were stained with anti-CD8 and <t>anti-IFN-γ,</t> then analyzed on a FACScalibur machine (A). PSGR-derived peptide-specific T cells were co-incubated with LNCaP cells alone in medium, or with LNCaP cells in the presence of either anti-HLA-I mAb (W6/32), HLA-II mAb or a control mAb (anti-CD19 mAb). After 4 hours of incubation, the cytotoxicity against LNCaP was determined by the LDH assay (B). Data from B are plotted as means ± SD. Results are representative of three independent experiments. * P
    Anti Human Ifn γ, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti human ifn γ/product/Thermo Fisher
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti human ifn γ - by Bioz Stars, 2021-05
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    99
    Thermo Fisher ifn γ
    A Subpharmacologic Dose of Sorafenib Triggers mCAR T Cell Activation in the Presence of Macrophages (A and B) Expression levels of IL12 mRNA in TAMs (A) and IL12 protein in the TME (B). Eight days after Hepa1-6-chGPC3 tumor implantation, mice were treated with vehicle or 7.5 mg/kg sorafenib for 5 days. On day 14, tumors were harvested for further detection (n = 3). TAMs were isolated with anti-F4/80 microbeads, and IL12 mRNA in TAMs was evaluated via qPCR (n = 3). Tumor lysates were assayed for IL12 via ELISA. (C) Sorafenib triggered IL12 secretion in BMDMs. BMDMs were treated with different concentrations of sorafenib and LPS (10 ng/mL) for 24 h. IL12 secretion into culture supernatants was assayed via ELISA. The data are representative of three independent experiments. (D) Cytotoxicity of sorafenib to BMDMs. The cell viability of BMDMs was shown after treatment with the indicated concentrations of sorafenib in the presence or absence of LPS (10 ng/mL) for 24 h. The data are representative of three independent experiments. (E) Transwell coculture of mCAR T cells with BMDMs. BMDMs were treated with sorafenib and LPS (10 ng/mL) for 8 h. Then, mCAR T cells and tumor cells were added to the upper chamber for another 16 h for coculture with sorafenib-treated BMDMs. <t>IFN-γ</t> expression in mCAR T cells was assayed by flow cytometry. (F) Blocking experiment. Sorafenib (2 μM) and anti-IL12 antibody (5 μg/mL) were added to the coculture system as mentioned in (E). Each data point reflects the mean ± SEM of triplicate experiments. Significance of findings was defined as follows: ns, not significant; p > 0.05; *p
    Ifn γ, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ifn γ/product/Thermo Fisher
    Average 99 stars, based on 1 article reviews
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    ifn γ - by Bioz Stars, 2021-05
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    97
    Thermo Fisher gamma interferon ifn γ
    Cytomix induces iNOS mRNA in A549 cells. A549 cells were treated for 2 or 24 h with 10 ng each of IL-1, TNF-α, and <t>IFN-γ/ml.</t> Total RNA was extracted from cell lysates, and multiplex RT-PCR was conducted with two primer sets in a single PCR (iNOS primers plus 18S rRNA primers and competimers as an invariant endogenous control). PCR products were resolved by electrophoresis on a 1.8% agarose-ethidium bromide gel. Lane 1, DNA markers; lane 2, untreated (2 h) A549; lane 3 and 5, Cytomix-treated (2 h) A549; lane 4, same as lane 3 but without reverse transcriptase; lane 6, untreated (24 h) A549; lanes 7 and 9, Cytomix-treated (24 h) A549; lane 8, same as lane 7 but without reverse transcriptase; lane 10, 18S rRNA control (495 bp); lane 11, iNOS RNA control (349 bp).
    Gamma Interferon Ifn γ, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/gamma interferon ifn γ/product/Thermo Fisher
    Average 97 stars, based on 1 article reviews
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    gamma interferon ifn γ - by Bioz Stars, 2021-05
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    IFN gamma Monoclonal Antibody for Flow
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    PSGR-derived peptide-induced T cell responses were CD8 + T cell dependent and restricted by HLA-I. PSGR-derived peptide-specific T cells were co-cultured with T2 cells pulsed with or without a given peptide in the presence of GolgiStop in a 48-well plate for 4 hrs at 37°C. Cells were stained with anti-CD8 and anti-IFN-γ, then analyzed on a FACScalibur machine (A). PSGR-derived peptide-specific T cells were co-incubated with LNCaP cells alone in medium, or with LNCaP cells in the presence of either anti-HLA-I mAb (W6/32), HLA-II mAb or a control mAb (anti-CD19 mAb). After 4 hours of incubation, the cytotoxicity against LNCaP was determined by the LDH assay (B). Data from B are plotted as means ± SD. Results are representative of three independent experiments. * P

    Journal: PLoS ONE

    Article Title: Identification of Prostate-Specific G-Protein Coupled Receptor as a Tumor Antigen Recognized by CD8+ T Cells for Cancer Immunotherapy

    doi: 10.1371/journal.pone.0045756

    Figure Lengend Snippet: PSGR-derived peptide-induced T cell responses were CD8 + T cell dependent and restricted by HLA-I. PSGR-derived peptide-specific T cells were co-cultured with T2 cells pulsed with or without a given peptide in the presence of GolgiStop in a 48-well plate for 4 hrs at 37°C. Cells were stained with anti-CD8 and anti-IFN-γ, then analyzed on a FACScalibur machine (A). PSGR-derived peptide-specific T cells were co-incubated with LNCaP cells alone in medium, or with LNCaP cells in the presence of either anti-HLA-I mAb (W6/32), HLA-II mAb or a control mAb (anti-CD19 mAb). After 4 hours of incubation, the cytotoxicity against LNCaP was determined by the LDH assay (B). Data from B are plotted as means ± SD. Results are representative of three independent experiments. * P

    Article Snippet: Briefly, 96-well ELISPOT plates (Millipore; Bedford, MA, USA) were coated overnight at 4°C with 7.5 µg/mL anti-human IFN-γ (Pierce Biotechnology; Rockford, IL, USA).

    Techniques: Derivative Assay, Cell Culture, Staining, Incubation, Lactate Dehydrogenase Assay

    PSGR-derived peptides induced peptide-specific T cells. The recognition of T2 cells pre-loaded with titrated concentrations of peptides (0–20 µg/ml) by expanded PSGR peptide-specific T cells was tested by ELISA assay (A). The expanded PSGR3 T cells (B and E), PSGR4 T cells (C and F) and PSGR14 T cells (D and G) were respectively co-incubated with T2 cells (1×10 4 cells/well) alone in complete medium (CM), or with T2 cells pre-loaded with either a corresponding peptide (5 µg/mL) or a control peptide as a negative control. Cells were incubated for 18 −24 hours, the IFN-γ secretion in the supernatant was determined by ELISA assay (B, C and D). IFN-γ spot-forming cells (SFC) were enumerated by ELISPOT assay (E, F and G). Data are plotted as means ± SD. Results are representative of at least three independent experiments. * P

    Journal: PLoS ONE

    Article Title: Identification of Prostate-Specific G-Protein Coupled Receptor as a Tumor Antigen Recognized by CD8+ T Cells for Cancer Immunotherapy

    doi: 10.1371/journal.pone.0045756

    Figure Lengend Snippet: PSGR-derived peptides induced peptide-specific T cells. The recognition of T2 cells pre-loaded with titrated concentrations of peptides (0–20 µg/ml) by expanded PSGR peptide-specific T cells was tested by ELISA assay (A). The expanded PSGR3 T cells (B and E), PSGR4 T cells (C and F) and PSGR14 T cells (D and G) were respectively co-incubated with T2 cells (1×10 4 cells/well) alone in complete medium (CM), or with T2 cells pre-loaded with either a corresponding peptide (5 µg/mL) or a control peptide as a negative control. Cells were incubated for 18 −24 hours, the IFN-γ secretion in the supernatant was determined by ELISA assay (B, C and D). IFN-γ spot-forming cells (SFC) were enumerated by ELISPOT assay (E, F and G). Data are plotted as means ± SD. Results are representative of at least three independent experiments. * P

    Article Snippet: Briefly, 96-well ELISPOT plates (Millipore; Bedford, MA, USA) were coated overnight at 4°C with 7.5 µg/mL anti-human IFN-γ (Pierce Biotechnology; Rockford, IL, USA).

    Techniques: Derivative Assay, Enzyme-linked Immunosorbent Assay, Incubation, Negative Control, Enzyme-linked Immunospot

    A Subpharmacologic Dose of Sorafenib Triggers mCAR T Cell Activation in the Presence of Macrophages (A and B) Expression levels of IL12 mRNA in TAMs (A) and IL12 protein in the TME (B). Eight days after Hepa1-6-chGPC3 tumor implantation, mice were treated with vehicle or 7.5 mg/kg sorafenib for 5 days. On day 14, tumors were harvested for further detection (n = 3). TAMs were isolated with anti-F4/80 microbeads, and IL12 mRNA in TAMs was evaluated via qPCR (n = 3). Tumor lysates were assayed for IL12 via ELISA. (C) Sorafenib triggered IL12 secretion in BMDMs. BMDMs were treated with different concentrations of sorafenib and LPS (10 ng/mL) for 24 h. IL12 secretion into culture supernatants was assayed via ELISA. The data are representative of three independent experiments. (D) Cytotoxicity of sorafenib to BMDMs. The cell viability of BMDMs was shown after treatment with the indicated concentrations of sorafenib in the presence or absence of LPS (10 ng/mL) for 24 h. The data are representative of three independent experiments. (E) Transwell coculture of mCAR T cells with BMDMs. BMDMs were treated with sorafenib and LPS (10 ng/mL) for 8 h. Then, mCAR T cells and tumor cells were added to the upper chamber for another 16 h for coculture with sorafenib-treated BMDMs. IFN-γ expression in mCAR T cells was assayed by flow cytometry. (F) Blocking experiment. Sorafenib (2 μM) and anti-IL12 antibody (5 μg/mL) were added to the coculture system as mentioned in (E). Each data point reflects the mean ± SEM of triplicate experiments. Significance of findings was defined as follows: ns, not significant; p > 0.05; *p

    Journal: Molecular Therapy

    Article Title: Combined Antitumor Effects of Sorafenib and GPC3-CAR T Cells in Mouse Models of Hepatocellular Carcinoma

    doi: 10.1016/j.ymthe.2019.04.020

    Figure Lengend Snippet: A Subpharmacologic Dose of Sorafenib Triggers mCAR T Cell Activation in the Presence of Macrophages (A and B) Expression levels of IL12 mRNA in TAMs (A) and IL12 protein in the TME (B). Eight days after Hepa1-6-chGPC3 tumor implantation, mice were treated with vehicle or 7.5 mg/kg sorafenib for 5 days. On day 14, tumors were harvested for further detection (n = 3). TAMs were isolated with anti-F4/80 microbeads, and IL12 mRNA in TAMs was evaluated via qPCR (n = 3). Tumor lysates were assayed for IL12 via ELISA. (C) Sorafenib triggered IL12 secretion in BMDMs. BMDMs were treated with different concentrations of sorafenib and LPS (10 ng/mL) for 24 h. IL12 secretion into culture supernatants was assayed via ELISA. The data are representative of three independent experiments. (D) Cytotoxicity of sorafenib to BMDMs. The cell viability of BMDMs was shown after treatment with the indicated concentrations of sorafenib in the presence or absence of LPS (10 ng/mL) for 24 h. The data are representative of three independent experiments. (E) Transwell coculture of mCAR T cells with BMDMs. BMDMs were treated with sorafenib and LPS (10 ng/mL) for 8 h. Then, mCAR T cells and tumor cells were added to the upper chamber for another 16 h for coculture with sorafenib-treated BMDMs. IFN-γ expression in mCAR T cells was assayed by flow cytometry. (F) Blocking experiment. Sorafenib (2 μM) and anti-IL12 antibody (5 μg/mL) were added to the coculture system as mentioned in (E). Each data point reflects the mean ± SEM of triplicate experiments. Significance of findings was defined as follows: ns, not significant; p > 0.05; *p

    Article Snippet: For detection of CD107a and IFN-γ, T cells were fixed, permeabilized, and stained with antibodies against CD107a and IFN-γ (eBioscience).

    Techniques: Activation Assay, Expressing, Tumor Implantation, Mouse Assay, Isolation, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Blocking Assay

    Sorafenib Has No Significant Effect on the Activity of huCAR T Cells In Vitro (A) Schematic representation of the modular composition of GPC3-specific human CAR. 9F2scFv, GPC3-specific single chain (heavy and light) fragment variable; tm, transmembrane domain; huCD28-CD3ζ, combined intracellular human CD28 and CD3ζ signaling domain. (B) Modified T cells express 9F2-hu28z CAR on their surface, based on flow cytometry results. (C) CD107a expression in activated huCAR T cells in the presence of sorafenib. huCAR T cells were stimulated with PLC/PRF/5 cells at 1:1 ratio in the presence of 1, 5, or 10 μM sorafenib for 24 h. Then, CD107a expression in huCAR T cells was assayed by flow cytometry. (D) The effect of sorafenib on huCAR T cell proliferation. huCAR T cells were prelabeled with Cell Trace Violet dye and then incubated with PLC/PRF/5 cells at a 1:1 ratio in the presence of different concentrations of sorafenib for 48 h. The numbers gated indicate the dividing cell population. (E) Cytotoxicity of sorafenib-pretreated huCAR T cells against tumor cells. huCAR T cells were pretreated with various concentrations of sorafenib for 4 h and then cocultured with PLC/PRF/5 cells at varying effector:target (E:T) ratios for 4 h for cytotoxic assays using a standard nonradioactive cytotoxic assay kit. The data are representative of three independent experiments. (F–H) In vitro cytokine production of huCAR T cells. Briefly, 1 × 10 5 huCAR T cells were cultivated in 24-well plates precoated with GPC3 protein. After 24 h, culture supernatants were harvested and assayed for IL-2 (F), IFN-γ (G), and TNF-α (H) via cytometric bead array (CBA). (I) Comparison of the activities (CD107a, IL-2, IFN-γ, and TNF-α) of mCAR and huCAR T cells after treatment with 10 μM sorafenib. All data are presented as the mean ± SEM of triplicate experiments, unless otherwise noted. Significance of findings was defined as follows: ns, not significant; p > 0.05; *p

    Journal: Molecular Therapy

    Article Title: Combined Antitumor Effects of Sorafenib and GPC3-CAR T Cells in Mouse Models of Hepatocellular Carcinoma

    doi: 10.1016/j.ymthe.2019.04.020

    Figure Lengend Snippet: Sorafenib Has No Significant Effect on the Activity of huCAR T Cells In Vitro (A) Schematic representation of the modular composition of GPC3-specific human CAR. 9F2scFv, GPC3-specific single chain (heavy and light) fragment variable; tm, transmembrane domain; huCD28-CD3ζ, combined intracellular human CD28 and CD3ζ signaling domain. (B) Modified T cells express 9F2-hu28z CAR on their surface, based on flow cytometry results. (C) CD107a expression in activated huCAR T cells in the presence of sorafenib. huCAR T cells were stimulated with PLC/PRF/5 cells at 1:1 ratio in the presence of 1, 5, or 10 μM sorafenib for 24 h. Then, CD107a expression in huCAR T cells was assayed by flow cytometry. (D) The effect of sorafenib on huCAR T cell proliferation. huCAR T cells were prelabeled with Cell Trace Violet dye and then incubated with PLC/PRF/5 cells at a 1:1 ratio in the presence of different concentrations of sorafenib for 48 h. The numbers gated indicate the dividing cell population. (E) Cytotoxicity of sorafenib-pretreated huCAR T cells against tumor cells. huCAR T cells were pretreated with various concentrations of sorafenib for 4 h and then cocultured with PLC/PRF/5 cells at varying effector:target (E:T) ratios for 4 h for cytotoxic assays using a standard nonradioactive cytotoxic assay kit. The data are representative of three independent experiments. (F–H) In vitro cytokine production of huCAR T cells. Briefly, 1 × 10 5 huCAR T cells were cultivated in 24-well plates precoated with GPC3 protein. After 24 h, culture supernatants were harvested and assayed for IL-2 (F), IFN-γ (G), and TNF-α (H) via cytometric bead array (CBA). (I) Comparison of the activities (CD107a, IL-2, IFN-γ, and TNF-α) of mCAR and huCAR T cells after treatment with 10 μM sorafenib. All data are presented as the mean ± SEM of triplicate experiments, unless otherwise noted. Significance of findings was defined as follows: ns, not significant; p > 0.05; *p

    Article Snippet: For detection of CD107a and IFN-γ, T cells were fixed, permeabilized, and stained with antibodies against CD107a and IFN-γ (eBioscience).

    Techniques: Activity Assay, In Vitro, Modification, Flow Cytometry, Expressing, Planar Chromatography, Incubation, Crocin Bleaching Assay

    High-Dose, but Not Low-Dose, Sorafenib Inhibits the Function of GPC3-mCAR T Cells In Vitro (A) Sorafenib inhibits CD107a expression and IFN-γ production in mouse CAR T cells. GPC3-mCAR T cells (1 × 10 5 cells per well) were stimulated with Hepa1-6-chGPC3 at a 1:1 ratio in the presence of 1, 5, or 10 μM sorafenib for 24 h. After cell surface and intracellular staining, the percentages of mCAR T cells expressing the relevant markers was determined via multicolor flow cytometry. (B) Sorafenib inhibits mCAR T cell proliferation. GPC3-mCAR T cells were labeled with Cell Trace Violet, and then, 1 × 10 5 Cell Trace–labeled mCAR T cells were incubated with 1 × 10 5 Hepa1-6-chGPC3 cells in the presence of various concentrations of sorafenib for 24 h. The number of gated cells indicate the dividing cell population. (C) Reduced cytotoxicity was observed after incubation of sorafenib-pretreated mCAR T cells with Hepa1-6-chGPC3 cells at various effector:target (E:T) ratios for 4 h, determined by a standard nonradioactive cytotoxic assay. (D–F) In vitro cytokine production of mCAR T cells. Briefly, 1 × 10 5 mCAR T cells were cultivated in 24-well plates precoated with GPC3 protein. After 24 h, culture supernatants were harvested and assayed for IL-2 (D), IFN-γ (E), and TNF-α (F) via cytometric bead array (CBA). All data are presented as the mean ± SEM of triplicate experiments. Significance of findings was defined as follows: ns, not significant; p > 0.05; *p

    Journal: Molecular Therapy

    Article Title: Combined Antitumor Effects of Sorafenib and GPC3-CAR T Cells in Mouse Models of Hepatocellular Carcinoma

    doi: 10.1016/j.ymthe.2019.04.020

    Figure Lengend Snippet: High-Dose, but Not Low-Dose, Sorafenib Inhibits the Function of GPC3-mCAR T Cells In Vitro (A) Sorafenib inhibits CD107a expression and IFN-γ production in mouse CAR T cells. GPC3-mCAR T cells (1 × 10 5 cells per well) were stimulated with Hepa1-6-chGPC3 at a 1:1 ratio in the presence of 1, 5, or 10 μM sorafenib for 24 h. After cell surface and intracellular staining, the percentages of mCAR T cells expressing the relevant markers was determined via multicolor flow cytometry. (B) Sorafenib inhibits mCAR T cell proliferation. GPC3-mCAR T cells were labeled with Cell Trace Violet, and then, 1 × 10 5 Cell Trace–labeled mCAR T cells were incubated with 1 × 10 5 Hepa1-6-chGPC3 cells in the presence of various concentrations of sorafenib for 24 h. The number of gated cells indicate the dividing cell population. (C) Reduced cytotoxicity was observed after incubation of sorafenib-pretreated mCAR T cells with Hepa1-6-chGPC3 cells at various effector:target (E:T) ratios for 4 h, determined by a standard nonradioactive cytotoxic assay. (D–F) In vitro cytokine production of mCAR T cells. Briefly, 1 × 10 5 mCAR T cells were cultivated in 24-well plates precoated with GPC3 protein. After 24 h, culture supernatants were harvested and assayed for IL-2 (D), IFN-γ (E), and TNF-α (F) via cytometric bead array (CBA). All data are presented as the mean ± SEM of triplicate experiments. Significance of findings was defined as follows: ns, not significant; p > 0.05; *p

    Article Snippet: For detection of CD107a and IFN-γ, T cells were fixed, permeabilized, and stained with antibodies against CD107a and IFN-γ (eBioscience).

    Techniques: In Vitro, Expressing, Staining, Flow Cytometry, Labeling, Incubation, Crocin Bleaching Assay

    A Subpharmacologic Dose of Sorafenib Augments the Antitumor Activity, Infiltration, and Cytokine Production of GPC3-mCAR T Cells In Vivo (A) In vivo experimental design. Hepa1-6-chGPC3 tumor cells (1 × 10 7 ) were subcutaneously injected into C57BL/6 mice and allowed to establish for 8 days. Mice were assigned to six experimental groups as indicated, vehicle or sorafenib at 7.5 (Sora7.5) or 30 (Sora30) mg/kg was administered via gavage for 10 days, and 2 × 10 6 GPC3-mCAR T cells were adoptively transferred i.v. on days 9 and 13. (B and C) Tumor growth (B) and survival (C) curves of treatment groups (n = 6). (D) Representative flow cytometry plots showing the frequencies of Cell Trace Violet prelabeled mCAR T cells in tumor-infiltrating CD45 + cells (E) and IFN-γ + cells in mCAR T cells (F) on day 11 (n = 4). (G) Treatment protocol. Six days after tumor inoculation, mice were divided into five experimental groups, as indicated, and then treated with 7.5 mg/kg (Sora7.5) sorafenib for 5 days. Then, 2 × 10 6 mCAR or untransduced (UTD) T cells were injected i.v. on day 7. (H) Tumor growth curves (n = 6). The results are expressed as the mean ± SEM. Significance of findings was defined as follows: ns, not significant; p > 0.05; *p

    Journal: Molecular Therapy

    Article Title: Combined Antitumor Effects of Sorafenib and GPC3-CAR T Cells in Mouse Models of Hepatocellular Carcinoma

    doi: 10.1016/j.ymthe.2019.04.020

    Figure Lengend Snippet: A Subpharmacologic Dose of Sorafenib Augments the Antitumor Activity, Infiltration, and Cytokine Production of GPC3-mCAR T Cells In Vivo (A) In vivo experimental design. Hepa1-6-chGPC3 tumor cells (1 × 10 7 ) were subcutaneously injected into C57BL/6 mice and allowed to establish for 8 days. Mice were assigned to six experimental groups as indicated, vehicle or sorafenib at 7.5 (Sora7.5) or 30 (Sora30) mg/kg was administered via gavage for 10 days, and 2 × 10 6 GPC3-mCAR T cells were adoptively transferred i.v. on days 9 and 13. (B and C) Tumor growth (B) and survival (C) curves of treatment groups (n = 6). (D) Representative flow cytometry plots showing the frequencies of Cell Trace Violet prelabeled mCAR T cells in tumor-infiltrating CD45 + cells (E) and IFN-γ + cells in mCAR T cells (F) on day 11 (n = 4). (G) Treatment protocol. Six days after tumor inoculation, mice were divided into five experimental groups, as indicated, and then treated with 7.5 mg/kg (Sora7.5) sorafenib for 5 days. Then, 2 × 10 6 mCAR or untransduced (UTD) T cells were injected i.v. on day 7. (H) Tumor growth curves (n = 6). The results are expressed as the mean ± SEM. Significance of findings was defined as follows: ns, not significant; p > 0.05; *p

    Article Snippet: For detection of CD107a and IFN-γ, T cells were fixed, permeabilized, and stained with antibodies against CD107a and IFN-γ (eBioscience).

    Techniques: Activity Assay, In Vivo, Injection, Mouse Assay, Flow Cytometry

    Cytomix induces iNOS mRNA in A549 cells. A549 cells were treated for 2 or 24 h with 10 ng each of IL-1, TNF-α, and IFN-γ/ml. Total RNA was extracted from cell lysates, and multiplex RT-PCR was conducted with two primer sets in a single PCR (iNOS primers plus 18S rRNA primers and competimers as an invariant endogenous control). PCR products were resolved by electrophoresis on a 1.8% agarose-ethidium bromide gel. Lane 1, DNA markers; lane 2, untreated (2 h) A549; lane 3 and 5, Cytomix-treated (2 h) A549; lane 4, same as lane 3 but without reverse transcriptase; lane 6, untreated (24 h) A549; lanes 7 and 9, Cytomix-treated (24 h) A549; lane 8, same as lane 7 but without reverse transcriptase; lane 10, 18S rRNA control (495 bp); lane 11, iNOS RNA control (349 bp).

    Journal: Infection and Immunity

    Article Title: Cytokine Responses to Group B Streptococci Induce Nitric Oxide Production in Respiratory Epithelial Cells

    doi: 10.1128/IAI.70.1.49-54.2002

    Figure Lengend Snippet: Cytomix induces iNOS mRNA in A549 cells. A549 cells were treated for 2 or 24 h with 10 ng each of IL-1, TNF-α, and IFN-γ/ml. Total RNA was extracted from cell lysates, and multiplex RT-PCR was conducted with two primer sets in a single PCR (iNOS primers plus 18S rRNA primers and competimers as an invariant endogenous control). PCR products were resolved by electrophoresis on a 1.8% agarose-ethidium bromide gel. Lane 1, DNA markers; lane 2, untreated (2 h) A549; lane 3 and 5, Cytomix-treated (2 h) A549; lane 4, same as lane 3 but without reverse transcriptase; lane 6, untreated (24 h) A549; lanes 7 and 9, Cytomix-treated (24 h) A549; lane 8, same as lane 7 but without reverse transcriptase; lane 10, 18S rRNA control (495 bp); lane 11, iNOS RNA control (349 bp).

    Article Snippet: Recombinant human interleukin-1-β (IL-1), tumor necrosis factor alpha (TNF-α), and gamma interferon (IFN-γ) were purchased from Biosource International (Camarillo, Calif.).

    Techniques: Multiplex Assay, Reverse Transcription Polymerase Chain Reaction, Polymerase Chain Reaction, Electrophoresis

    Evaluation of the anti-tumor and anti-metastatic potential of ESC+VPA treatment with respect to tumor stem cells (A) hESC+VPA treatment reduced tumor volume (mm 3 ) compared to controls at day 27 after challenge with mammospheres (n=5 per group). (B) mESC+VPA treatment reduced tumor volume (mm 3 ) compared to controls at day 27 after challenge with mammospheres (n=5 per group). (C) Bioluminescence imaging of hESC+VPA and mESC+VPA treatment groups compared to control group. (D) Lung metastases in treated (hESCs+VPA, mESCs+VPA) and control mice, as shown by total flux (photons/second) (E) Bioluminescence images of the surgically removed lungs of control and hESC+VPA-treated mice. (F) Quantification by flow cytometry of CD4 + and CD8 + tumor-infiltrating lymphocytes from treated and untreated mice. (G) Dot plot images of flow cytometry results, showing an increase in CD4 + and CD8 + T cells in mice treated with hESC+VPA compared to the control group. (H) Quantification of Gr1 + CD11b + cell populations in tumors of mice treated with ESCs+VPA compared to the control group. (I) Dot plot images of flow cytometry results, showing the double Gr1 + CD11b + cell populations in tumors in control mice and in mice treated with hESC+VPA. (J) A negative correlation was found between the secretion of IFN-γ (pg/mL) by tumor-infiltrating lymphocytes and tumor burden. (K) Quantification of IFN-gamma (pg/mL) secretion by tumor-infiltrating lymphocytes in treated and control mice.

    Journal: bioRxiv

    Article Title: Pharmacologically modified pluripotent stem cell-based cancer vaccines with anti-metastatic potential

    doi: 10.1101/2020.05.27.118471

    Figure Lengend Snippet: Evaluation of the anti-tumor and anti-metastatic potential of ESC+VPA treatment with respect to tumor stem cells (A) hESC+VPA treatment reduced tumor volume (mm 3 ) compared to controls at day 27 after challenge with mammospheres (n=5 per group). (B) mESC+VPA treatment reduced tumor volume (mm 3 ) compared to controls at day 27 after challenge with mammospheres (n=5 per group). (C) Bioluminescence imaging of hESC+VPA and mESC+VPA treatment groups compared to control group. (D) Lung metastases in treated (hESCs+VPA, mESCs+VPA) and control mice, as shown by total flux (photons/second) (E) Bioluminescence images of the surgically removed lungs of control and hESC+VPA-treated mice. (F) Quantification by flow cytometry of CD4 + and CD8 + tumor-infiltrating lymphocytes from treated and untreated mice. (G) Dot plot images of flow cytometry results, showing an increase in CD4 + and CD8 + T cells in mice treated with hESC+VPA compared to the control group. (H) Quantification of Gr1 + CD11b + cell populations in tumors of mice treated with ESCs+VPA compared to the control group. (I) Dot plot images of flow cytometry results, showing the double Gr1 + CD11b + cell populations in tumors in control mice and in mice treated with hESC+VPA. (J) A negative correlation was found between the secretion of IFN-γ (pg/mL) by tumor-infiltrating lymphocytes and tumor burden. (K) Quantification of IFN-gamma (pg/mL) secretion by tumor-infiltrating lymphocytes in treated and control mice.

    Article Snippet: IFN-gamma was quantified using the IFN-gamma kit (eBioscience) following the manufacturer’s protocol.

    Techniques: Imaging, Mouse Assay, Flow Cytometry