anti-ifn-γ Search Results


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  • 93
    BioLegend anti ifn γ
    The proportion and suppressive function of Treg cells are maintained in Smad4 tKO NOD mice SLCs were isolated from Smad4 tKO and WT NOD mice at 12 weeks of age. ( A ) The proportion and ( B ) absolute number of Treg (CD4 + CD25 + Foxp3 + T) cells were determined by flow cytometry. C. The expression of Foxp3 mRNA was analyzed by qRT-PCR. D. Representative histogram plots of suppression assay of Treg cells. CFSE-labelled effector T cells (CD4 + CD25 − T; Teff) from WT NOD mice were stimulated with anti-CD3/CD28-coated beads for 72 h in the presence of Treg (CD4 + CD25 + T) cells from WT or Smad4 tKO NOD mice at various ratios (left margin). Proliferation of Teff cells was assessed by flow cytometry. E. The percentage of cells undergoing the indicated number of divisions when the Treg:Teff cell ratio was 1:1. Values are means ± SD ( n = 3-4/group). ( F ) Total RNA was isolated from Treg cells and the expression of mRNA for <t>IFN-γ,</t> IL-10 and TGF-β was analyzed by qRT-PCR. Values are means ± SD ( n = 8-9/group), * P
    Anti Ifn γ, supplied by BioLegend, used in various techniques. Bioz Stars score: 93/100, based on 1834 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    R&D Systems anti ifn γ
    Interferon <t>(IFN)-λ</t> 2/3 exerts a potent antiviral effect against influenza A virus (IAV) infection in the mouse lung. Non-asthmatic mice ( N = 5) and asthmatic mice ( N = 5) were infected with 213 pfu IAV WS/33 (H1N1), and body weight (A) and survival rate [ (B) , N = 20] were assessed until 14 dpi. The mRNA levels of TNF-α, IL-1β, and Ccl7 (C) and the levels of secreted IFN-λ 2/3 and <t>IFN-γ</t> (D) in the lung and bronchoalveolar lavage fluid were determined. Asthmatic mice were infected with 213 pfu IAV WS/33 (H1N1) and treated with recombinant IFN-λ 2/3 (IFN-λ 2 : 1 μg and IFN-λ 3 : 1 μg/30 μl) ( N = 5). As a control, the diluent phosphate-buffered saline was used ( N = 5). Survival rate [ (E) , N = 20], hematoxylin/eosin-stained lung histologic findings (F) , and viral titer (G) were determined at 14 dpi. Micrographs are representative of lung sections from five mice. Polymerase chain reaction, plaque assay, and enzyme-linked immunosorbent assay results are presented as mean ± SD from five independent experiments (* p
    Anti Ifn γ, supplied by R&D Systems, used in various techniques. Bioz Stars score: 94/100, based on 629 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Chromogenix anti ifn γ
    Detection of Th1 cytokines adjacent to gastric epithelial cells during infection with H. pylori . Biopsy specimens from infected and uninfected subjects were stained with antibodies recognizing human <t>IFN-γ</t> or TNF-α as described in Materials and Methods. Whereas few positive cells were detected in uninfected subjects, cytokine-positive cells were observed within or adjacent to the gastric epithelium (arrows). These images are representative of eight different subjects.
    Anti Ifn γ, supplied by Chromogenix, used in various techniques. Bioz Stars score: 92/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Becton Dickinson anti ifn γ
    Flow cytometry analysis of lymphocyte populations in mesenteric lymph nodes following ileitis induction. Lymphocyte populations were isolated from mesenteric lymph nodes (MLN) derived from C57BL/6 wildtype (WT) and TLR-9 deficient (TLR-9-/-) mice before (naïve) and seven days following ileitis induction (ILE) and analyzed by flow cytometry. Frequencies of CD4+ cells producing <t>IFN-γ</t> ( right panel ), CD4+ CD3+ cells ( left panel ), as well as CD69+ CD4+ cells ( middle panel ), are indicated in representative FACS plots (in %). Data shown are representative for three independent experiments.
    Anti Ifn γ, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 92/100, based on 4124 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Mab Technologies anti ifn γ
    Results of the <t>IFN-γ</t> and IL-4 responses to PMA/ionomycin activation of PBMC from Saimiri sciureus samples. Samples from nine Saimiri (U1, R1, O13, O17 Q3, J3, J7 B11 and N5) and one clinically healthy human donor (HUM) were cultured in absence or presence of PMA/ionomycin (PMA/Ion) for 18 hrs. The results of ELISPOT for (A) IFN-γ and (B) IL-4 are presented as the mean of the number of SFU/10 6 cells ± SD. Mean of OD values obtained ELISA assays for (C) IFN-γ and (D) IL-4 ± SD.
    Anti Ifn γ, supplied by Mab Technologies, used in various techniques. Bioz Stars score: 93/100, based on 344 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ImmunoTools anti ifn γ
    Results of the <t>IFN-γ</t> and IL-4 responses to PMA/ionomycin activation of PBMC from Saimiri sciureus samples. Samples from nine Saimiri (U1, R1, O13, O17 Q3, J3, J7 B11 and N5) and one clinically healthy human donor (HUM) were cultured in absence or presence of PMA/ionomycin (PMA/Ion) for 18 hrs. The results of ELISPOT for (A) IFN-γ and (B) IL-4 are presented as the mean of the number of SFU/10 6 cells ± SD. Mean of OD values obtained ELISA assays for (C) IFN-γ and (D) IL-4 ± SD.
    Anti Ifn γ, supplied by ImmunoTools, used in various techniques. Bioz Stars score: 92/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    2Bio Ltd anti ifn γ
    Il17a −/− Il17f −/− dKO mice exhibit reduced T H 2 cell responses against intranasal allergens. A, Representative FACS plot of CD4 + T cells from the lung. FSC , Forward scatter. B and C, Percentages (Fig 1, B ) and absolute numbers (Fig 1, C ) of CD4 + T-cell subsets. D, Levels of <t>IFN-γ,</t> IL-4, and IL-5 in BAL fluid. The graph shows means ± SEMs. * P
    Anti Ifn γ, supplied by 2Bio Ltd, used in various techniques. Bioz Stars score: 90/100, based on 61 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    Santa Cruz Biotechnology anti interferon γ
    Representative Western blots (A) and bar graphs showing densitometric quantifications of the inflammatory markers interferon gamma <t>(IFN-γ)</t> (B) , inter-cellular adhesion molecule-1 (ICAM-1) (C) , and interleukin 17 (IL-17) (D) at 72 hours after cICH-induction
    Anti Interferon γ, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 89/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Bio-Rad anti ifn γ
    Antigen-specific whole blood assay. ( A) Time-course of IL-17A and <t>IFN-γ</t> production by whole blood cultured with OVA. The blood of the 10 responder cows (taken 45 days after the first immunization) was cultured in the presence of OVA (10 μg/mL) for 1 to 4 days in 96-well microplates. For each cow triplicate wells were used for each incubation time. Results are median values and quartiles (Q1; Q3). (B) The effect of magnetic depletion of CD4+ cells on the production of IL-17A and IFN-γ in the antigen-specific whole blood assay. Results obtained by stimulating blood samples from responder cows 15 days after the booster immunization are expressed as the percentage of the production by CD4+-depleted PBMC relative to cytokine production by un processed PBMC (100%). The viability of cells was not altered by the magnetic cell separation.
    Anti Ifn γ, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 92/100, based on 23 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Pharmingen anti ifn γ
    Production of <t>IFN-γ</t> and IL-4 by AgI/II-specific cells from superficial lymph nodes (A) and spleens (B and C) of BALB/c mice immunized i.n. with AgI/II with or without CT, LT-IIa, or LT-IIb. Cells were stimulated in vitro with AgI/II at the concentrations shown for 4 days. Data shown are the arithmetic mean values ± standard deviations ( n = 4) as determined by cytokine-specific ELISA. In panels A and B, ∗, ∗∗, and ∗∗∗ indicate significant differences at P
    Anti Ifn γ, supplied by Pharmingen, used in various techniques. Bioz Stars score: 92/100, based on 314 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Thermo Fisher anti ifn γ
    ILDR2-mFc treatment inhibits myelin epitope-specific epitope spreading in the R-EAE model. ( A ) On day 76 postdisease induction, five mice from each treatment group were analyzed for recall responses to spread epitopes, via injection of 10 μg of PLP 178–191 in one ear and MBP 84–104 into the opposite ear. The level of ear swelling was assayed 24 h postchallenge. The data are presented as the mean net ear swelling. ( B ) Recall responses were also carried on splenocytes. On day 76 postdisease induction, total splenocytes were collected from five representative mice from each treatment group and activated in the presence of OVA 323–339 , PLP 139–151 , PLP 178–191 , or MBP 84–104 (20 μg/ml). Cells were pulsed with 1 mCi of tritiated thymidine at 24 h and harvested 72 h postculture set up. Cell proliferation in recall responses was also carried out using splenocytes from day-45 mice ( C ) or <t>IFN-γ,</t> IL-17, and IL-4 secretion was analyzed by LiquiChip following 72 h of culture ( D–F ). One representative experiment of two or three independent experiments is presented for each experimental set. * p
    Anti Ifn γ, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 2696 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Tonbo Biosciences anti ifn γ
    ILDR2-mFc treatment inhibits myelin epitope-specific epitope spreading in the R-EAE model. ( A ) On day 76 postdisease induction, five mice from each treatment group were analyzed for recall responses to spread epitopes, via injection of 10 μg of PLP 178–191 in one ear and MBP 84–104 into the opposite ear. The level of ear swelling was assayed 24 h postchallenge. The data are presented as the mean net ear swelling. ( B ) Recall responses were also carried on splenocytes. On day 76 postdisease induction, total splenocytes were collected from five representative mice from each treatment group and activated in the presence of OVA 323–339 , PLP 139–151 , PLP 178–191 , or MBP 84–104 (20 μg/ml). Cells were pulsed with 1 mCi of tritiated thymidine at 24 h and harvested 72 h postculture set up. Cell proliferation in recall responses was also carried out using splenocytes from day-45 mice ( C ) or <t>IFN-γ,</t> IL-17, and IL-4 secretion was analyzed by LiquiChip following 72 h of culture ( D–F ). One representative experiment of two or three independent experiments is presented for each experimental set. * p
    Anti Ifn γ, supplied by Tonbo Biosciences, used in various techniques. Bioz Stars score: 91/100, based on 20 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Miltenyi Biotec anti ifn γ
    ABCC3 expressed by CD56 dim CD16 + NK cells is an indicator of better patient prognosis. ( A – C ). ( A ) Representative dot plots showing that CD56 dim CD16 + NK cells express ( B ) high levels of ABCC3, ( C ) and CD56 dim CD16 + ABCC3 + NK cells express <t>IFN-γ.</t> ( D ) Time course of frequency of NK cells expressing ABCC3 measured by flow cytometry (* p
    Anti Ifn γ, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 90/100, based on 74 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Beckman Coulter anti ifn γ
    Quantitation of soluble factors in LGL and healthy controls by ELISA and FACS. (A and C–F), Determination of expression levels of <t>IFN-γ,</t> IL-18/IGIF, MCP1/CCL2, IL-8/CXCL8, and IP10/CXCL10 in plasma samples from 25 to 29 LGL patients and
    Anti Ifn γ, supplied by Beckman Coulter, used in various techniques. Bioz Stars score: 92/100, based on 24 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    BioExpress anti ifn γ
    Quantitation of soluble factors in LGL and healthy controls by ELISA and FACS. (A and C–F), Determination of expression levels of <t>IFN-γ,</t> IL-18/IGIF, MCP1/CCL2, IL-8/CXCL8, and IP10/CXCL10 in plasma samples from 25 to 29 LGL patients and
    Anti Ifn γ, supplied by BioExpress, used in various techniques. Bioz Stars score: 92/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Harlan Laboratories anti ifn γ
    Quantitation of soluble factors in LGL and healthy controls by ELISA and FACS. (A and C–F), Determination of expression levels of <t>IFN-γ,</t> IL-18/IGIF, MCP1/CCL2, IL-8/CXCL8, and IP10/CXCL10 in plasma samples from 25 to 29 LGL patients and
    Anti Ifn γ, supplied by Harlan Laboratories, used in various techniques. Bioz Stars score: 92/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Diaclone anti ifn γ
    Combined cytokine profiles in response to M. leprae . Production of <t>IFN-γ</t> ( A ), IP-10 ( B ) and IL-10 ( C ) determined by ELISA, in response to medium (-), PHA, M. leprae WCS or the M. leprae -unique protein ML2478 in 24 h WBA for Ethiopian leprosy patients (n = 11: 2 BT (○) and 9 BL (•), and healthy endemic controls (EC; n = 12; □). For comparison between BT and BL, significant differences were found for M. leprae WCS (Mlep) induced IFN-γ responses (p = 0.036) and ML2478 induced IL-10 responses (p = 0.035). ( D ): IP-10/IL-10 ratios are depicted for unstimulated samples after 24 h {LP (•) and EC (□)} or after 1 h WBA {LP (▵) and EC (▾)}. ( E ): Anti-PGL-I antibodies for BL (○) and BT (•) patients were detected by ELISA using natural disaccharide of PGL-I linked to HSA [31] (ND-O-HSA). Optical density (OD 450 ) readings were performed using 1∶800 serum dilutions. Median values per group are indicated by horizontal lines. The cut-off for positivity is indicated by the dashed horizontal line.
    Anti Ifn γ, supplied by Diaclone, used in various techniques. Bioz Stars score: 92/100, based on 23 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Bioworld Antibodies anti ifn γ
    Synergistic effect of macrophage and natural killer (NK) cells on hepatic apoptosis induced by lupus immunoglobulin G (IgG). (A) Western blot showing the levels of TNF-α and <t>IFN-γ</t> in the liver from C57BL/6 mice with intrahepatic injection of the same dose of systemic lupus erythematosus (SLE) and normal serum, SLE, and normal IgG. (B) Flow cytometry analysis of NK cell activation in the liver from B6 lpr (25 w) and age-matched control mice using anti-NK1.1, anti-CD3e and anti CD69 antibodies. The result is representative of three independent experiments, n = 6 mice per group/experiment. (C) Flow cytometry analysis of NK cell activation in the liver from C57BL/6 mice with intrahepatic injection of SLE-serum (treated for 72 h), SLE-IgG (treated for 48 h), or phosphate-buffered saline (treated 48 h) using anti-NK1.1, anti-CD3e and anti CD69 antibodies (the result is representative of three independent experiments, n = 5 mice per group/experiment). (D) Severity of liver inflammation 48 h after intrahepatic injection of SLE-IgG in C57BL/6 mice with or without NK depletion using anti-ASGM1 antibody treatment. * p
    Anti Ifn γ, supplied by Bioworld Antibodies, used in various techniques. Bioz Stars score: 92/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    U-CyTech anti ifn γ ab
    Synergistic effect of macrophage and natural killer (NK) cells on hepatic apoptosis induced by lupus immunoglobulin G (IgG). (A) Western blot showing the levels of TNF-α and <t>IFN-γ</t> in the liver from C57BL/6 mice with intrahepatic injection of the same dose of systemic lupus erythematosus (SLE) and normal serum, SLE, and normal IgG. (B) Flow cytometry analysis of NK cell activation in the liver from B6 lpr (25 w) and age-matched control mice using anti-NK1.1, anti-CD3e and anti CD69 antibodies. The result is representative of three independent experiments, n = 6 mice per group/experiment. (C) Flow cytometry analysis of NK cell activation in the liver from C57BL/6 mice with intrahepatic injection of SLE-serum (treated for 72 h), SLE-IgG (treated for 48 h), or phosphate-buffered saline (treated 48 h) using anti-NK1.1, anti-CD3e and anti CD69 antibodies (the result is representative of three independent experiments, n = 5 mice per group/experiment). (D) Severity of liver inflammation 48 h after intrahepatic injection of SLE-IgG in C57BL/6 mice with or without NK depletion using anti-ASGM1 antibody treatment. * p
    Anti Ifn γ Ab, supplied by U-CyTech, used in various techniques. Bioz Stars score: 91/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Leinco Technologies anti ifn γ
    Immune checkpoint blockade induces expansion of Th1 CD4 + T cells that express T-bet and <t>IFN-γ.</t> ( A ) Tumor-infiltrating lymphocytes (TILs) were analyzed in day 14 tumors by flow cytometry. ICB was initiated on day 9 and repeated on day 12. Data are shown as mean ± SD from n = 9 tumors aggregated from 2 experiments. ( B ) CD4 + T cell subtype analysis in TILs by flow cytometry. ICB was initiated on day 9 and repeated on day 12. Data are shown as mean ± SD from n = 9 tumors aggregated from 2 experiments. ( C ) IFN-γ expression in intratumoral T-bet + CD4 + T cells after 4 hours of PMA/ionomycin stimulation in vitro. Data are plotted as mean ± SD from n = 5–8 tumors. ( D ) CD4 + T cells treated as in C and costained for IFN-γ and T-bet. ( E ) Similar to B , showing the percentage of T-bet + CD4 + TILs that stained for Ki67. ( F ) Percentage of Ki67 + cells in CD4 + or CD8 + T cells in dLNs on day 14 (mean ± SD). n for schematic of gating strategies. All statistical analysis by Student’s t test. NS > 0.05, *** P
    Anti Ifn γ, supplied by Leinco Technologies, used in various techniques. Bioz Stars score: 92/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    The proportion and suppressive function of Treg cells are maintained in Smad4 tKO NOD mice SLCs were isolated from Smad4 tKO and WT NOD mice at 12 weeks of age. ( A ) The proportion and ( B ) absolute number of Treg (CD4 + CD25 + Foxp3 + T) cells were determined by flow cytometry. C. The expression of Foxp3 mRNA was analyzed by qRT-PCR. D. Representative histogram plots of suppression assay of Treg cells. CFSE-labelled effector T cells (CD4 + CD25 − T; Teff) from WT NOD mice were stimulated with anti-CD3/CD28-coated beads for 72 h in the presence of Treg (CD4 + CD25 + T) cells from WT or Smad4 tKO NOD mice at various ratios (left margin). Proliferation of Teff cells was assessed by flow cytometry. E. The percentage of cells undergoing the indicated number of divisions when the Treg:Teff cell ratio was 1:1. Values are means ± SD ( n = 3-4/group). ( F ) Total RNA was isolated from Treg cells and the expression of mRNA for IFN-γ, IL-10 and TGF-β was analyzed by qRT-PCR. Values are means ± SD ( n = 8-9/group), * P

    Journal: Oncotarget

    Article Title: Smad4 in T cells plays a protective role in the development of autoimmune Sjögren's syndrome in the nonobese diabetic mouse

    doi: 10.18632/oncotarget.13437

    Figure Lengend Snippet: The proportion and suppressive function of Treg cells are maintained in Smad4 tKO NOD mice SLCs were isolated from Smad4 tKO and WT NOD mice at 12 weeks of age. ( A ) The proportion and ( B ) absolute number of Treg (CD4 + CD25 + Foxp3 + T) cells were determined by flow cytometry. C. The expression of Foxp3 mRNA was analyzed by qRT-PCR. D. Representative histogram plots of suppression assay of Treg cells. CFSE-labelled effector T cells (CD4 + CD25 − T; Teff) from WT NOD mice were stimulated with anti-CD3/CD28-coated beads for 72 h in the presence of Treg (CD4 + CD25 + T) cells from WT or Smad4 tKO NOD mice at various ratios (left margin). Proliferation of Teff cells was assessed by flow cytometry. E. The percentage of cells undergoing the indicated number of divisions when the Treg:Teff cell ratio was 1:1. Values are means ± SD ( n = 3-4/group). ( F ) Total RNA was isolated from Treg cells and the expression of mRNA for IFN-γ, IL-10 and TGF-β was analyzed by qRT-PCR. Values are means ± SD ( n = 8-9/group), * P

    Article Snippet: INC.), mouse-IL-23 (10 ng/ml; Biolegend), human-TGF-β (5 ng/ml), anti-IL4 (10 μg/ml; Biolegend) and anti-IFN-γ (10 μg/ml; Biolegend) for Th17; mouse IL-2 (10 ng/ml), human-TGF-β (2 ng/ml), anti-IL4 (10 μg/ml) and anti-IFN-γ (10 μg/ml) for inducible Treg (iTreg) cell polarization.

    Techniques: Mouse Assay, Isolation, Flow Cytometry, Cytometry, Expressing, Quantitative RT-PCR, Suppression Assay

    Expression of inflammatory cytokines is increased in exocrine glands from Smad4 tKO NOD mice ( A and B ) Total RNA and ( C ) tissue lysates were prepared from lacrimal and salivary glands of Smad4 tKO NOD and WT NOD mice at 12 weeks of age. The expression of mRNA for various cytokines ( A ) and transcription factors ( B ) was analyzed by qRT-PCR. Values are expressed as the relative fold-change as compared with WT. ( C ) IFN-γ and IL-17 production was measured by ELISA. Data are mean ± SD ( n = 6~9/group). * P

    Journal: Oncotarget

    Article Title: Smad4 in T cells plays a protective role in the development of autoimmune Sjögren's syndrome in the nonobese diabetic mouse

    doi: 10.18632/oncotarget.13437

    Figure Lengend Snippet: Expression of inflammatory cytokines is increased in exocrine glands from Smad4 tKO NOD mice ( A and B ) Total RNA and ( C ) tissue lysates were prepared from lacrimal and salivary glands of Smad4 tKO NOD and WT NOD mice at 12 weeks of age. The expression of mRNA for various cytokines ( A ) and transcription factors ( B ) was analyzed by qRT-PCR. Values are expressed as the relative fold-change as compared with WT. ( C ) IFN-γ and IL-17 production was measured by ELISA. Data are mean ± SD ( n = 6~9/group). * P

    Article Snippet: INC.), mouse-IL-23 (10 ng/ml; Biolegend), human-TGF-β (5 ng/ml), anti-IL4 (10 μg/ml; Biolegend) and anti-IFN-γ (10 μg/ml; Biolegend) for Th17; mouse IL-2 (10 ng/ml), human-TGF-β (2 ng/ml), anti-IL4 (10 μg/ml) and anti-IFN-γ (10 μg/ml) for inducible Treg (iTreg) cell polarization.

    Techniques: Expressing, Mouse Assay, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

    Inflammatory cytokine-expressing T cells are increased in SLCs from Smad4 tKO NOD mice A. - B. Total RNA was isolated from SLCs from Smad4 tKO and WT NOD mice at 12 weeks of age. The expression of mRNA for various ( A ) cytokines and ( B ) transcription factors was analyzed by quantitative real-time PCR. Values are expressed as the relative fold-change as compared with WT ( n = 3-4/group). C. SLCs from Smad4 tKO and WT NOD mice at 12 weeks of age were stimulated with PMA (10 ng/ml) and ionomycin (500 ng/ml) for 12 h. The proportions of T cells producing IFN-γ, IL-4 and IL-17 were measured by FACS analysis ( n = 5/group). Data are mean ± SD. * P

    Journal: Oncotarget

    Article Title: Smad4 in T cells plays a protective role in the development of autoimmune Sjögren's syndrome in the nonobese diabetic mouse

    doi: 10.18632/oncotarget.13437

    Figure Lengend Snippet: Inflammatory cytokine-expressing T cells are increased in SLCs from Smad4 tKO NOD mice A. - B. Total RNA was isolated from SLCs from Smad4 tKO and WT NOD mice at 12 weeks of age. The expression of mRNA for various ( A ) cytokines and ( B ) transcription factors was analyzed by quantitative real-time PCR. Values are expressed as the relative fold-change as compared with WT ( n = 3-4/group). C. SLCs from Smad4 tKO and WT NOD mice at 12 weeks of age were stimulated with PMA (10 ng/ml) and ionomycin (500 ng/ml) for 12 h. The proportions of T cells producing IFN-γ, IL-4 and IL-17 were measured by FACS analysis ( n = 5/group). Data are mean ± SD. * P

    Article Snippet: INC.), mouse-IL-23 (10 ng/ml; Biolegend), human-TGF-β (5 ng/ml), anti-IL4 (10 μg/ml; Biolegend) and anti-IFN-γ (10 μg/ml; Biolegend) for Th17; mouse IL-2 (10 ng/ml), human-TGF-β (2 ng/ml), anti-IL4 (10 μg/ml) and anti-IFN-γ (10 μg/ml) for inducible Treg (iTreg) cell polarization.

    Techniques: Expressing, Mouse Assay, Isolation, Real-time Polymerase Chain Reaction, FACS

    Identification of CD4 + Th1 interferon γ (IFN-γ) + cells specific for RSV G (183–195) peptide induced by vaccine inoculation. Lung mononuclear cells were isolated from mice vaccinated twice and sacrificed 5 days after RSV challenge. Isolated cells were stimulated with G peptide (183–195: WAICKRIPNKKPG) for 5 hours in vitro . Cells were stained with anti-CD3 antibody, anti-CD4 antibody, and anti-CD44 antibody and then fixed. Permeabilized cells were stained with anti-IFN-γ antibody and analyzed by flow cytometry. (A) The dot plots in the gated area represent CD4 + CD44 + T cells (Th1 cells) that can secrete IFN-γ. (B) The percentage of CD4 + CD44 + T cells (Th1 cells) that can secrete IFN-γ for each vaccine-immunized group. Data are expressed as mean±standard deviation (n=5/group). Statistically significant values are marked with an asterisk. *** p

    Journal: Clinical and Experimental Vaccine Research

    Article Title: Vaccine containing G protein fragment and recombinant baculovirus expressing M2 protein induces protective immunity to respiratory syncytial virus

    doi: 10.7774/cevr.2019.8.1.43

    Figure Lengend Snippet: Identification of CD4 + Th1 interferon γ (IFN-γ) + cells specific for RSV G (183–195) peptide induced by vaccine inoculation. Lung mononuclear cells were isolated from mice vaccinated twice and sacrificed 5 days after RSV challenge. Isolated cells were stimulated with G peptide (183–195: WAICKRIPNKKPG) for 5 hours in vitro . Cells were stained with anti-CD3 antibody, anti-CD4 antibody, and anti-CD44 antibody and then fixed. Permeabilized cells were stained with anti-IFN-γ antibody and analyzed by flow cytometry. (A) The dot plots in the gated area represent CD4 + CD44 + T cells (Th1 cells) that can secrete IFN-γ. (B) The percentage of CD4 + CD44 + T cells (Th1 cells) that can secrete IFN-γ for each vaccine-immunized group. Data are expressed as mean±standard deviation (n=5/group). Statistically significant values are marked with an asterisk. *** p

    Article Snippet: The permeabilized cells were stained with anti-IFN-γ antibody (XMG 1.2, BioLegend) at RT for 30 minutes in the dark.

    Techniques: Isolation, Mouse Assay, In Vitro, Staining, Flow Cytometry, Cytometry, Standard Deviation

    Comparison of CD8 + interferon γ (IFN-γ) + T cells specific for RSV M2 (82-90) peptide after vaccination. Mice were challenged with RSV after 2 vaccinations and lung mononuclear cells were harvested at day 5 to identify CD8 + CD44 + T cells capable of secreting IFN-γ. The harvested lung mononuclear cells were stimulated with M2 peptide (82-90: SYIGSINNI) for 5 hours in vitro . Stimulated lung cells were stained with anti-CD3, anti-CD8, and anti-CD44 antibodies and then fixed. Permeabilized cells were stained with anti-IFN-γ antibody and analyzed by flow cytometry. (A) The dot plots in the gated area represent the CD8 + CD44 + T cells that can secrete IFN-γ. (B) The percentage of CD8 + CD44 + T cells that can secrete IFN-γ for each vaccine-immunized group. Data are expressed as mean± standard deviation (n=5/group). Statistically significant values are marked with an asterisk. *** p

    Journal: Clinical and Experimental Vaccine Research

    Article Title: Vaccine containing G protein fragment and recombinant baculovirus expressing M2 protein induces protective immunity to respiratory syncytial virus

    doi: 10.7774/cevr.2019.8.1.43

    Figure Lengend Snippet: Comparison of CD8 + interferon γ (IFN-γ) + T cells specific for RSV M2 (82-90) peptide after vaccination. Mice were challenged with RSV after 2 vaccinations and lung mononuclear cells were harvested at day 5 to identify CD8 + CD44 + T cells capable of secreting IFN-γ. The harvested lung mononuclear cells were stimulated with M2 peptide (82-90: SYIGSINNI) for 5 hours in vitro . Stimulated lung cells were stained with anti-CD3, anti-CD8, and anti-CD44 antibodies and then fixed. Permeabilized cells were stained with anti-IFN-γ antibody and analyzed by flow cytometry. (A) The dot plots in the gated area represent the CD8 + CD44 + T cells that can secrete IFN-γ. (B) The percentage of CD8 + CD44 + T cells that can secrete IFN-γ for each vaccine-immunized group. Data are expressed as mean± standard deviation (n=5/group). Statistically significant values are marked with an asterisk. *** p

    Article Snippet: The permeabilized cells were stained with anti-IFN-γ antibody (XMG 1.2, BioLegend) at RT for 30 minutes in the dark.

    Techniques: Mouse Assay, In Vitro, Staining, Flow Cytometry, Cytometry, Standard Deviation

    Interferon (IFN)-λ 2/3 exerts a potent antiviral effect against influenza A virus (IAV) infection in the mouse lung. Non-asthmatic mice ( N = 5) and asthmatic mice ( N = 5) were infected with 213 pfu IAV WS/33 (H1N1), and body weight (A) and survival rate [ (B) , N = 20] were assessed until 14 dpi. The mRNA levels of TNF-α, IL-1β, and Ccl7 (C) and the levels of secreted IFN-λ 2/3 and IFN-γ (D) in the lung and bronchoalveolar lavage fluid were determined. Asthmatic mice were infected with 213 pfu IAV WS/33 (H1N1) and treated with recombinant IFN-λ 2/3 (IFN-λ 2 : 1 μg and IFN-λ 3 : 1 μg/30 μl) ( N = 5). As a control, the diluent phosphate-buffered saline was used ( N = 5). Survival rate [ (E) , N = 20], hematoxylin/eosin-stained lung histologic findings (F) , and viral titer (G) were determined at 14 dpi. Micrographs are representative of lung sections from five mice. Polymerase chain reaction, plaque assay, and enzyme-linked immunosorbent assay results are presented as mean ± SD from five independent experiments (* p

    Journal: Frontiers in Immunology

    Article Title: Initial Influenza Virus Replication Can Be Limited in Allergic Asthma Through Rapid Induction of Type III Interferons in Respiratory Epithelium

    doi: 10.3389/fimmu.2018.00986

    Figure Lengend Snippet: Interferon (IFN)-λ 2/3 exerts a potent antiviral effect against influenza A virus (IAV) infection in the mouse lung. Non-asthmatic mice ( N = 5) and asthmatic mice ( N = 5) were infected with 213 pfu IAV WS/33 (H1N1), and body weight (A) and survival rate [ (B) , N = 20] were assessed until 14 dpi. The mRNA levels of TNF-α, IL-1β, and Ccl7 (C) and the levels of secreted IFN-λ 2/3 and IFN-γ (D) in the lung and bronchoalveolar lavage fluid were determined. Asthmatic mice were infected with 213 pfu IAV WS/33 (H1N1) and treated with recombinant IFN-λ 2/3 (IFN-λ 2 : 1 μg and IFN-λ 3 : 1 μg/30 μl) ( N = 5). As a control, the diluent phosphate-buffered saline was used ( N = 5). Survival rate [ (E) , N = 20], hematoxylin/eosin-stained lung histologic findings (F) , and viral titer (G) were determined at 14 dpi. Micrographs are representative of lung sections from five mice. Polymerase chain reaction, plaque assay, and enzyme-linked immunosorbent assay results are presented as mean ± SD from five independent experiments (* p

    Article Snippet: Anti-IFN-λ2/3 (cat number: MAB1789) and anti-IFN-γ (cat number: MAB4851) neutralizing antibodies and isotype-control antibodies (rat IgG) were purchased from R & D Systems.

    Techniques: Infection, Mouse Assay, Recombinant, Staining, Polymerase Chain Reaction, Plaque Assay, Enzyme-linked Immunosorbent Assay

    Acute influenza A virus (IAV) infection is aggravated in asthmatic mice administered neutralizing antibodies against interferon (IFN)-λ 2/3 and IFN-γ. Asthmatic mice with isotype-control (rat IgG) antibodies were infected with 213 pfu IAV WS/33 (H1N1) ( N = 5) and treated with neutralizing antibodies against IFN-λ 2/3 ( N = 5) or IFN-γ ( N = 5). Hematoxylin/eosin (H E)-stained micrographs were generated from lung sections obtained from IAV-infected asthmatic mice on day 7 (A) . Viral titer (B) and secreted IFN level (C,D) were determined by plaque assay and enzyme-linked immunosorbent assay. Histological assessment was performed using periodic acid Schiff (PAS)-stained lung sections of asthmatic mice ( N = 5) (E) . Viral titer (F) and Th2 cytokine levels (G) were assessed using a plaque assay and a multiplex assay at 0 ( N = 5), 1 ( N = 5), 3 ( N = 5), 5 ( N = 5), and 7 ( N = 5) dpi. Micrographs shown are representative of lung sections from five mice. Polymerase chain reaction, plaque assay, and multiplex assay results are presented as mean ± SD from five independent experiments (* p

    Journal: Frontiers in Immunology

    Article Title: Initial Influenza Virus Replication Can Be Limited in Allergic Asthma Through Rapid Induction of Type III Interferons in Respiratory Epithelium

    doi: 10.3389/fimmu.2018.00986

    Figure Lengend Snippet: Acute influenza A virus (IAV) infection is aggravated in asthmatic mice administered neutralizing antibodies against interferon (IFN)-λ 2/3 and IFN-γ. Asthmatic mice with isotype-control (rat IgG) antibodies were infected with 213 pfu IAV WS/33 (H1N1) ( N = 5) and treated with neutralizing antibodies against IFN-λ 2/3 ( N = 5) or IFN-γ ( N = 5). Hematoxylin/eosin (H E)-stained micrographs were generated from lung sections obtained from IAV-infected asthmatic mice on day 7 (A) . Viral titer (B) and secreted IFN level (C,D) were determined by plaque assay and enzyme-linked immunosorbent assay. Histological assessment was performed using periodic acid Schiff (PAS)-stained lung sections of asthmatic mice ( N = 5) (E) . Viral titer (F) and Th2 cytokine levels (G) were assessed using a plaque assay and a multiplex assay at 0 ( N = 5), 1 ( N = 5), 3 ( N = 5), 5 ( N = 5), and 7 ( N = 5) dpi. Micrographs shown are representative of lung sections from five mice. Polymerase chain reaction, plaque assay, and multiplex assay results are presented as mean ± SD from five independent experiments (* p

    Article Snippet: Anti-IFN-λ2/3 (cat number: MAB1789) and anti-IFN-γ (cat number: MAB4851) neutralizing antibodies and isotype-control antibodies (rat IgG) were purchased from R & D Systems.

    Techniques: Infection, Mouse Assay, Staining, Generated, Plaque Assay, Enzyme-linked Immunosorbent Assay, Multiplex Assay, Polymerase Chain Reaction

    Detection of Th1 cytokines adjacent to gastric epithelial cells during infection with H. pylori . Biopsy specimens from infected and uninfected subjects were stained with antibodies recognizing human IFN-γ or TNF-α as described in Materials and Methods. Whereas few positive cells were detected in uninfected subjects, cytokine-positive cells were observed within or adjacent to the gastric epithelium (arrows). These images are representative of eight different subjects.

    Journal: Infection and Immunity

    Article Title: Helicobacter pylori Modulates Lymphoepithelial Cell Interactions Leading to Epithelial Cell Damage through Fas/Fas Ligand Interactions

    doi:

    Figure Lengend Snippet: Detection of Th1 cytokines adjacent to gastric epithelial cells during infection with H. pylori . Biopsy specimens from infected and uninfected subjects were stained with antibodies recognizing human IFN-γ or TNF-α as described in Materials and Methods. Whereas few positive cells were detected in uninfected subjects, cytokine-positive cells were observed within or adjacent to the gastric epithelium (arrows). These images are representative of eight different subjects.

    Article Snippet: The slides were then incubated overnight at 4°C with anti-TNF-α (MAB11; PharMingen, San Diego, Calif.) or anti-IFN-γ (DIK-1; Chromogenix, Molndal, Sweden) monoclonal antibodies at a concentration of 5 μg/ml.

    Techniques: Infection, Staining

    Flow cytometry analysis of lymphocyte populations in mesenteric lymph nodes following ileitis induction. Lymphocyte populations were isolated from mesenteric lymph nodes (MLN) derived from C57BL/6 wildtype (WT) and TLR-9 deficient (TLR-9-/-) mice before (naïve) and seven days following ileitis induction (ILE) and analyzed by flow cytometry. Frequencies of CD4+ cells producing IFN-γ ( right panel ), CD4+ CD3+ cells ( left panel ), as well as CD69+ CD4+ cells ( middle panel ), are indicated in representative FACS plots (in %). Data shown are representative for three independent experiments.

    Journal: Gut Pathogens

    Article Title: The impact of Toll-like-receptor-9 on intestinal microbiota composition and extra-intestinal sequelae in experimental Toxoplasma gondii induced ileitis

    doi: 10.1186/1757-4749-6-19

    Figure Lengend Snippet: Flow cytometry analysis of lymphocyte populations in mesenteric lymph nodes following ileitis induction. Lymphocyte populations were isolated from mesenteric lymph nodes (MLN) derived from C57BL/6 wildtype (WT) and TLR-9 deficient (TLR-9-/-) mice before (naïve) and seven days following ileitis induction (ILE) and analyzed by flow cytometry. Frequencies of CD4+ cells producing IFN-γ ( right panel ), CD4+ CD3+ cells ( left panel ), as well as CD69+ CD4+ cells ( middle panel ), are indicated in representative FACS plots (in %). Data shown are representative for three independent experiments.

    Article Snippet: Stainings and cell sorting with anti-CD3 (clone 145-2C11, isotype hamster IgG1; BD Pharmingen), anti-CD4 (clone GK1.5 isotype rat IgG2b; BD Pharmingen), anti-CD69 (clone H1.2 F3, isotype Hamster IgG1*, Lamda 3; BD Pharmingen), and anti-IFN-γ (clone 145-2C11, isotype hamster IgG1; BD Pharmingen) were performed.

    Techniques: Flow Cytometry, Cytometry, Isolation, Derivative Assay, Mouse Assay, FACS

    Systemic and extra-intestinal pro-inflammatory cytokine responses following ileitis induction. Serum (A) IL-6 and (B) IFN-γ, splenic (C) TNF-α and (D) IFN-γ as well as hepatic (E) TNF-α and (F) IL-6 protein concentrations were determined in respective ex vivo biopsies derived from C57BL/6 wildtype (WT; black circles) and TLR-9 deficient (TLR-9-/-; white circles) mice seven days following ileitis induction (ILE). Naïve mice served as negative controls (N). Numbers of analyzed mice (in parentheses), means (black bars) and levels of significance ( P -values) as compared to the respective groups (determined by Mann–Whitney- U test) are indicated. Data shown were pooled from three independent experiments.

    Journal: Gut Pathogens

    Article Title: The impact of Toll-like-receptor-9 on intestinal microbiota composition and extra-intestinal sequelae in experimental Toxoplasma gondii induced ileitis

    doi: 10.1186/1757-4749-6-19

    Figure Lengend Snippet: Systemic and extra-intestinal pro-inflammatory cytokine responses following ileitis induction. Serum (A) IL-6 and (B) IFN-γ, splenic (C) TNF-α and (D) IFN-γ as well as hepatic (E) TNF-α and (F) IL-6 protein concentrations were determined in respective ex vivo biopsies derived from C57BL/6 wildtype (WT; black circles) and TLR-9 deficient (TLR-9-/-; white circles) mice seven days following ileitis induction (ILE). Naïve mice served as negative controls (N). Numbers of analyzed mice (in parentheses), means (black bars) and levels of significance ( P -values) as compared to the respective groups (determined by Mann–Whitney- U test) are indicated. Data shown were pooled from three independent experiments.

    Article Snippet: Stainings and cell sorting with anti-CD3 (clone 145-2C11, isotype hamster IgG1; BD Pharmingen), anti-CD4 (clone GK1.5 isotype rat IgG2b; BD Pharmingen), anti-CD69 (clone H1.2 F3, isotype Hamster IgG1*, Lamda 3; BD Pharmingen), and anti-IFN-γ (clone 145-2C11, isotype hamster IgG1; BD Pharmingen) were performed.

    Techniques: Ex Vivo, Derivative Assay, Mouse Assay, MANN-WHITNEY

    Ileal cytokine secretion following ileitis induction. (A) IFN-γ, (B) TNF-α, (C) nitric oxide, (D) IL-6, and (E) IL-10 protein levels were determined in ex vivo ileal biopsies derived from C57BL/6 wildtype (WT; black circles) and TLR-9 deficient (TLR-9-/-; white circles) mice seven days following ileitis induction (ILE) as described in methods. Naïve mice served as negative controls (N). Numbers of analyzed mice (in parentheses), means (black bars) and levels of significance ( P -values) as compared to the respective groups (determined by Mann–Whitney- U test) are indicated. Data shown were pooled from three independent experiments.

    Journal: Gut Pathogens

    Article Title: The impact of Toll-like-receptor-9 on intestinal microbiota composition and extra-intestinal sequelae in experimental Toxoplasma gondii induced ileitis

    doi: 10.1186/1757-4749-6-19

    Figure Lengend Snippet: Ileal cytokine secretion following ileitis induction. (A) IFN-γ, (B) TNF-α, (C) nitric oxide, (D) IL-6, and (E) IL-10 protein levels were determined in ex vivo ileal biopsies derived from C57BL/6 wildtype (WT; black circles) and TLR-9 deficient (TLR-9-/-; white circles) mice seven days following ileitis induction (ILE) as described in methods. Naïve mice served as negative controls (N). Numbers of analyzed mice (in parentheses), means (black bars) and levels of significance ( P -values) as compared to the respective groups (determined by Mann–Whitney- U test) are indicated. Data shown were pooled from three independent experiments.

    Article Snippet: Stainings and cell sorting with anti-CD3 (clone 145-2C11, isotype hamster IgG1; BD Pharmingen), anti-CD4 (clone GK1.5 isotype rat IgG2b; BD Pharmingen), anti-CD69 (clone H1.2 F3, isotype Hamster IgG1*, Lamda 3; BD Pharmingen), and anti-IFN-γ (clone 145-2C11, isotype hamster IgG1; BD Pharmingen) were performed.

    Techniques: Ex Vivo, Derivative Assay, Mouse Assay, MANN-WHITNEY

    Pro-inflammatory cytokine responses in mesenteric lymph nodes following ileitis induction. (A) IFN-γ, B) TNF-α, and (C) nitric oxide protein concentrations were determined in ex vivo biopsies of mesenteric lymph nodes (MLN) derived from C57BL/6 wildtype (WT; black circles) and TLR-9 deficient (TLR-9-/-; white circles) mice seven days following ileitis induction (ILE). Naïve mice served as negative controls (N). Numbers of analyzed mice (in parentheses), means (black bars) and levels of significance ( P -values) as compared to the respective groups (determined by Mann–Whitney- U test) are indicated. Data shown were pooled from three independent experiments.

    Journal: Gut Pathogens

    Article Title: The impact of Toll-like-receptor-9 on intestinal microbiota composition and extra-intestinal sequelae in experimental Toxoplasma gondii induced ileitis

    doi: 10.1186/1757-4749-6-19

    Figure Lengend Snippet: Pro-inflammatory cytokine responses in mesenteric lymph nodes following ileitis induction. (A) IFN-γ, B) TNF-α, and (C) nitric oxide protein concentrations were determined in ex vivo biopsies of mesenteric lymph nodes (MLN) derived from C57BL/6 wildtype (WT; black circles) and TLR-9 deficient (TLR-9-/-; white circles) mice seven days following ileitis induction (ILE). Naïve mice served as negative controls (N). Numbers of analyzed mice (in parentheses), means (black bars) and levels of significance ( P -values) as compared to the respective groups (determined by Mann–Whitney- U test) are indicated. Data shown were pooled from three independent experiments.

    Article Snippet: Stainings and cell sorting with anti-CD3 (clone 145-2C11, isotype hamster IgG1; BD Pharmingen), anti-CD4 (clone GK1.5 isotype rat IgG2b; BD Pharmingen), anti-CD69 (clone H1.2 F3, isotype Hamster IgG1*, Lamda 3; BD Pharmingen), and anti-IFN-γ (clone 145-2C11, isotype hamster IgG1; BD Pharmingen) were performed.

    Techniques: Ex Vivo, Derivative Assay, Mouse Assay, MANN-WHITNEY

    Cerebral histopathological changes following acute ileitis induction. C57BL/6 wildtype (WT; black circles) and TLR-9 deficient (TLR-9-/-; white circles) mice were perorally infected with 100 cysts of T. gondii ME49 strain on day 0 to induce acute ileitis. Naïve mice served as negative controls (N). Intracerebral (A) IFN-γ, (B) TNF-α, and (C) IL-6 mRNA expression levels were measured in ex vivo brain biopsies applying quantitative RT-PCR. Numbers of analyzed mice (in parentheses), means (black bars), and levels of significance ( P -values) determined by Mann–Whitney- U test are indicated. Data shown were pooled from three independent experiments.

    Journal: Gut Pathogens

    Article Title: The impact of Toll-like-receptor-9 on intestinal microbiota composition and extra-intestinal sequelae in experimental Toxoplasma gondii induced ileitis

    doi: 10.1186/1757-4749-6-19

    Figure Lengend Snippet: Cerebral histopathological changes following acute ileitis induction. C57BL/6 wildtype (WT; black circles) and TLR-9 deficient (TLR-9-/-; white circles) mice were perorally infected with 100 cysts of T. gondii ME49 strain on day 0 to induce acute ileitis. Naïve mice served as negative controls (N). Intracerebral (A) IFN-γ, (B) TNF-α, and (C) IL-6 mRNA expression levels were measured in ex vivo brain biopsies applying quantitative RT-PCR. Numbers of analyzed mice (in parentheses), means (black bars), and levels of significance ( P -values) determined by Mann–Whitney- U test are indicated. Data shown were pooled from three independent experiments.

    Article Snippet: Stainings and cell sorting with anti-CD3 (clone 145-2C11, isotype hamster IgG1; BD Pharmingen), anti-CD4 (clone GK1.5 isotype rat IgG2b; BD Pharmingen), anti-CD69 (clone H1.2 F3, isotype Hamster IgG1*, Lamda 3; BD Pharmingen), and anti-IFN-γ (clone 145-2C11, isotype hamster IgG1; BD Pharmingen) were performed.

    Techniques: Mouse Assay, Infection, Expressing, Ex Vivo, Quantitative RT-PCR, MANN-WHITNEY

    Memory response characteristics of total CD8 + T cells and H-2D d /p18 tetramer-positive CD8 + T cells following rVac-gp160 boost immunization of mice previously administered an anti-IFN-γ Ab in association with rAd-gp140 prime immunization. (A) H-2D

    Journal: Clinical and Vaccine Immunology : CVI

    Article Title: Anti-Gamma Interferon Antibodies Enhance the Immunogenicity of Recombinant Adenovirus Vectors ▿

    doi: 10.1128/CVI.05180-11

    Figure Lengend Snippet: Memory response characteristics of total CD8 + T cells and H-2D d /p18 tetramer-positive CD8 + T cells following rVac-gp160 boost immunization of mice previously administered an anti-IFN-γ Ab in association with rAd-gp140 prime immunization. (A) H-2D

    Article Snippet: Threefold serial dilutions of recombinant IFN-γ protein (human; BD Biosciences) and/or the anti-IFN-γ Ab (human, clone B27; BD Biosciences) were performed in 96-well plates in Dulbecco's modified Eagle medium (DMEM) supplemented with 10% FBS (final volume, 50 μl).

    Techniques: Mouse Assay

    Expression of CD62L on the surfaces of total CD8 + T cells and H-2D d /p18 tetramer-positive CD8 + T cells following in vivo anti-IFN-γ Ab administration to rAd-gp140-immunized mice. Shown are the percentages of total CD8 + T cells (left) and H-2D

    Journal: Clinical and Vaccine Immunology : CVI

    Article Title: Anti-Gamma Interferon Antibodies Enhance the Immunogenicity of Recombinant Adenovirus Vectors ▿

    doi: 10.1128/CVI.05180-11

    Figure Lengend Snippet: Expression of CD62L on the surfaces of total CD8 + T cells and H-2D d /p18 tetramer-positive CD8 + T cells following in vivo anti-IFN-γ Ab administration to rAd-gp140-immunized mice. Shown are the percentages of total CD8 + T cells (left) and H-2D

    Article Snippet: Threefold serial dilutions of recombinant IFN-γ protein (human; BD Biosciences) and/or the anti-IFN-γ Ab (human, clone B27; BD Biosciences) were performed in 96-well plates in Dulbecco's modified Eagle medium (DMEM) supplemented with 10% FBS (final volume, 50 μl).

    Techniques: Expressing, In Vivo, Mouse Assay

    Kinetics of rAd-gp140-induced p18-specific CD8 + T cell responses following in vivo anti-IFN-γ Ab administration. H-2D d /p18 tetramer-positive CD8 + T cells, shown as a percentage of total CD8 + T cells, were monitored in the PBMC of BALB/c mice inoculated

    Journal: Clinical and Vaccine Immunology : CVI

    Article Title: Anti-Gamma Interferon Antibodies Enhance the Immunogenicity of Recombinant Adenovirus Vectors ▿

    doi: 10.1128/CVI.05180-11

    Figure Lengend Snippet: Kinetics of rAd-gp140-induced p18-specific CD8 + T cell responses following in vivo anti-IFN-γ Ab administration. H-2D d /p18 tetramer-positive CD8 + T cells, shown as a percentage of total CD8 + T cells, were monitored in the PBMC of BALB/c mice inoculated

    Article Snippet: Threefold serial dilutions of recombinant IFN-γ protein (human; BD Biosciences) and/or the anti-IFN-γ Ab (human, clone B27; BD Biosciences) were performed in 96-well plates in Dulbecco's modified Eagle medium (DMEM) supplemented with 10% FBS (final volume, 50 μl).

    Techniques: In Vivo, Mouse Assay

    Results of the IFN-γ and IL-4 responses to PMA/ionomycin activation of PBMC from Saimiri sciureus samples. Samples from nine Saimiri (U1, R1, O13, O17 Q3, J3, J7 B11 and N5) and one clinically healthy human donor (HUM) were cultured in absence or presence of PMA/ionomycin (PMA/Ion) for 18 hrs. The results of ELISPOT for (A) IFN-γ and (B) IL-4 are presented as the mean of the number of SFU/10 6 cells ± SD. Mean of OD values obtained ELISA assays for (C) IFN-γ and (D) IL-4 ± SD.

    Journal: Malaria Journal

    Article Title: Molecular and immunological tools for the evaluation of the cellular immune response in the neotropical monkey Saimiri sciureus, a non-human primate model for malaria research

    doi: 10.1186/s12936-015-0688-1

    Figure Lengend Snippet: Results of the IFN-γ and IL-4 responses to PMA/ionomycin activation of PBMC from Saimiri sciureus samples. Samples from nine Saimiri (U1, R1, O13, O17 Q3, J3, J7 B11 and N5) and one clinically healthy human donor (HUM) were cultured in absence or presence of PMA/ionomycin (PMA/Ion) for 18 hrs. The results of ELISPOT for (A) IFN-γ and (B) IL-4 are presented as the mean of the number of SFU/10 6 cells ± SD. Mean of OD values obtained ELISA assays for (C) IFN-γ and (D) IL-4 ± SD.

    Article Snippet: However, only the anti-IFN-γ (clones mAb GZ-4 and 7-B6-1) human antibodies from Mabtech supplier facilitated the detection of this cytokine in both human and NHP cells (Table ).

    Techniques: Activation Assay, Cell Culture, Enzyme-linked Immunospot, Enzyme-linked Immunosorbent Assay

    Relative expression of cytokines in PBMC from Saimiri sciureus using TaqMan® Gene Expression Array Plates. The gene expression analysis of IFN-γ, IL-10, IL-12B, IL-17A, IL-1A, IL-2 and TNF from three Saimiri sciureus (S8, S34 and S36) cultured in the presence of PMA/ionomycin for 12 hours. The relative expression was considered as the expression of the corresponding genes in PBMC obtained from the healthy human donor.

    Journal: Malaria Journal

    Article Title: Molecular and immunological tools for the evaluation of the cellular immune response in the neotropical monkey Saimiri sciureus, a non-human primate model for malaria research

    doi: 10.1186/s12936-015-0688-1

    Figure Lengend Snippet: Relative expression of cytokines in PBMC from Saimiri sciureus using TaqMan® Gene Expression Array Plates. The gene expression analysis of IFN-γ, IL-10, IL-12B, IL-17A, IL-1A, IL-2 and TNF from three Saimiri sciureus (S8, S34 and S36) cultured in the presence of PMA/ionomycin for 12 hours. The relative expression was considered as the expression of the corresponding genes in PBMC obtained from the healthy human donor.

    Article Snippet: However, only the anti-IFN-γ (clones mAb GZ-4 and 7-B6-1) human antibodies from Mabtech supplier facilitated the detection of this cytokine in both human and NHP cells (Table ).

    Techniques: Expressing, Cell Culture

    Plots showing real-time PCR amplification using custom TaqMan® Gene Expression Assays. Samples from three Saimiri sciureus (S8, S36 and S34) and one healthy human donor (HUM) were cultured in absence (BR) or presence of PMA/ionomycin (PMA ION) for 12 h, 24 and 48 h. Delta Rn versus cycle amplification plots were obtained using primers and probes for (A) IL-4, (B) IL-5, (C) IL-6, (D) LTA, and (E) IFN-γ. The lines represent the threshold.

    Journal: Malaria Journal

    Article Title: Molecular and immunological tools for the evaluation of the cellular immune response in the neotropical monkey Saimiri sciureus, a non-human primate model for malaria research

    doi: 10.1186/s12936-015-0688-1

    Figure Lengend Snippet: Plots showing real-time PCR amplification using custom TaqMan® Gene Expression Assays. Samples from three Saimiri sciureus (S8, S36 and S34) and one healthy human donor (HUM) were cultured in absence (BR) or presence of PMA/ionomycin (PMA ION) for 12 h, 24 and 48 h. Delta Rn versus cycle amplification plots were obtained using primers and probes for (A) IL-4, (B) IL-5, (C) IL-6, (D) LTA, and (E) IFN-γ. The lines represent the threshold.

    Article Snippet: However, only the anti-IFN-γ (clones mAb GZ-4 and 7-B6-1) human antibodies from Mabtech supplier facilitated the detection of this cytokine in both human and NHP cells (Table ).

    Techniques: Real-time Polymerase Chain Reaction, Amplification, Expressing, Cell Culture

    Relative expression of cytokines in PBMC from Saimiri sciureus using TaqMan® Gene Expression Array Plates. Gene expression analysis of IFN-γ, IL-10, IL-12B, IL-17A, IL-1A, IL-2 and TNF in PBMC from two Saimiri sciureus (V17 and V31) cultured in the presence of PMA/ionomycin for 12 hours. The relative expression was determined as the expression of the corresponding genes in PBMC obtained from the healthy human donor.

    Journal: Malaria Journal

    Article Title: Molecular and immunological tools for the evaluation of the cellular immune response in the neotropical monkey Saimiri sciureus, a non-human primate model for malaria research

    doi: 10.1186/s12936-015-0688-1

    Figure Lengend Snippet: Relative expression of cytokines in PBMC from Saimiri sciureus using TaqMan® Gene Expression Array Plates. Gene expression analysis of IFN-γ, IL-10, IL-12B, IL-17A, IL-1A, IL-2 and TNF in PBMC from two Saimiri sciureus (V17 and V31) cultured in the presence of PMA/ionomycin for 12 hours. The relative expression was determined as the expression of the corresponding genes in PBMC obtained from the healthy human donor.

    Article Snippet: However, only the anti-IFN-γ (clones mAb GZ-4 and 7-B6-1) human antibodies from Mabtech supplier facilitated the detection of this cytokine in both human and NHP cells (Table ).

    Techniques: Expressing, Cell Culture

    Plots showing real-time PCR amplification using custom TaqMan® Gene Expression Assays. Samples from three Saimiri sciureus (24, P51 and 149) and one human health donor (HUM) were cultured in absence (BR) or presence of PMA/ionomycin (PMA ION) for 12 h. Delta Rn versus cycle amplification plots were obtained using primers and probes for (A) IL-4, (B) IL-5, (C) IL-6, (D) LTA, and (E) IFN-γ. The lines represent the threshold.

    Journal: Malaria Journal

    Article Title: Molecular and immunological tools for the evaluation of the cellular immune response in the neotropical monkey Saimiri sciureus, a non-human primate model for malaria research

    doi: 10.1186/s12936-015-0688-1

    Figure Lengend Snippet: Plots showing real-time PCR amplification using custom TaqMan® Gene Expression Assays. Samples from three Saimiri sciureus (24, P51 and 149) and one human health donor (HUM) were cultured in absence (BR) or presence of PMA/ionomycin (PMA ION) for 12 h. Delta Rn versus cycle amplification plots were obtained using primers and probes for (A) IL-4, (B) IL-5, (C) IL-6, (D) LTA, and (E) IFN-γ. The lines represent the threshold.

    Article Snippet: However, only the anti-IFN-γ (clones mAb GZ-4 and 7-B6-1) human antibodies from Mabtech supplier facilitated the detection of this cytokine in both human and NHP cells (Table ).

    Techniques: Real-time Polymerase Chain Reaction, Amplification, Expressing, Cell Culture

    Il17a −/− Il17f −/− dKO mice exhibit reduced T H 2 cell responses against intranasal allergens. A, Representative FACS plot of CD4 + T cells from the lung. FSC , Forward scatter. B and C, Percentages (Fig 1, B ) and absolute numbers (Fig 1, C ) of CD4 + T-cell subsets. D, Levels of IFN-γ, IL-4, and IL-5 in BAL fluid. The graph shows means ± SEMs. * P

    Journal: The Journal of allergy and clinical immunology

    Article Title: Concomitant suppression of TH2 and TH17 cell responses in allergic asthma by targeting retinoic acid receptor–related orphan receptor γt

    doi: 10.1016/j.jaci.2017.07.050

    Figure Lengend Snippet: Il17a −/− Il17f −/− dKO mice exhibit reduced T H 2 cell responses against intranasal allergens. A, Representative FACS plot of CD4 + T cells from the lung. FSC , Forward scatter. B and C, Percentages (Fig 1, B ) and absolute numbers (Fig 1, C ) of CD4 + T-cell subsets. D, Levels of IFN-γ, IL-4, and IL-5 in BAL fluid. The graph shows means ± SEMs. * P

    Article Snippet: For IFN-γ neutralization experiment, anti–IFN-γ (5 μg/mL, XMG1.2; Bio X Cell) was added also.

    Techniques: Mouse Assay, FACS

    RORγt-deficient mice exhibit reduced T H 2 and T H 17 responses against intranasal allergens. A, Representative FACS plot of lymphocytes from the lung. B and C, Percentages (Fig 2, B ) and absolute numbers (Fig 2, C ) of lung CD4 + T-cell subsets. D, Levels of IFN-γ, IL-4, IL-5, and IL-17A in BAL fluid. E, Absolute numbers of total cells, eosinophils (eo) , macrophages (mac) , lymphocytes (lym) , and neutrophils (neu) in BAL fluid. Data are representative of 3 independent experiments. The graph shows means ± SEMs. * P

    Journal: The Journal of allergy and clinical immunology

    Article Title: Concomitant suppression of TH2 and TH17 cell responses in allergic asthma by targeting retinoic acid receptor–related orphan receptor γt

    doi: 10.1016/j.jaci.2017.07.050

    Figure Lengend Snippet: RORγt-deficient mice exhibit reduced T H 2 and T H 17 responses against intranasal allergens. A, Representative FACS plot of lymphocytes from the lung. B and C, Percentages (Fig 2, B ) and absolute numbers (Fig 2, C ) of lung CD4 + T-cell subsets. D, Levels of IFN-γ, IL-4, IL-5, and IL-17A in BAL fluid. E, Absolute numbers of total cells, eosinophils (eo) , macrophages (mac) , lymphocytes (lym) , and neutrophils (neu) in BAL fluid. Data are representative of 3 independent experiments. The graph shows means ± SEMs. * P

    Article Snippet: For IFN-γ neutralization experiment, anti–IFN-γ (5 μg/mL, XMG1.2; Bio X Cell) was added also.

    Techniques: Mouse Assay, FACS

    Reduction of T H 2 cell responses by UA treatment is IFN-γ independent. A and B, Frequency (Fig 4, A ) and absolute number (Fig 4, B ) of BAL fluid CD4 + T-cell subsets. C, Absolute numbers of total cells, macrophages (mac) , eosinophils (eo) , neutrophils (neu) , and lymphocytes (lym) in BAL fluid. Data are representative of 3 independent experiments. The graph shows means ± SEMs. * P

    Journal: The Journal of allergy and clinical immunology

    Article Title: Concomitant suppression of TH2 and TH17 cell responses in allergic asthma by targeting retinoic acid receptor–related orphan receptor γt

    doi: 10.1016/j.jaci.2017.07.050

    Figure Lengend Snippet: Reduction of T H 2 cell responses by UA treatment is IFN-γ independent. A and B, Frequency (Fig 4, A ) and absolute number (Fig 4, B ) of BAL fluid CD4 + T-cell subsets. C, Absolute numbers of total cells, macrophages (mac) , eosinophils (eo) , neutrophils (neu) , and lymphocytes (lym) in BAL fluid. Data are representative of 3 independent experiments. The graph shows means ± SEMs. * P

    Article Snippet: For IFN-γ neutralization experiment, anti–IFN-γ (5 μg/mL, XMG1.2; Bio X Cell) was added also.

    Techniques:

    Immune status change in the tumour microenvironment post-virus treatments. B6 mice were inoculated i.p. with 5 × 10 5 MC38-luc cells and treated with PBS, vvDD, vvDD-IL-2-FG, or vvDD-IL-2-RG at 2 × 10 8 PFU per mouse 9 days post-tumour inoculation. Tumour-bearing mice were sacrificed 5 days post-treatment and primary tumours were collected and analysed by flow cytometry to determine CD4 + Foxp3 - ( a ) and CD8 + IFN-γ + T cells ( b ), exhausted CD8 + T cell ( c–f ), memory-phenotype T cells (CD8 + CD44 hi ) ( g ), regulatory T cells (CD4 + Foxp3 + ) ( h ), CD8/Treg ( i ), or by RT-qPCR to determine IFN-γ, granzyme B, perforin, CXCL9, TGF-β, CD105, and VEGF ( j–p ). Four to five mice were used for each treatment group and data are representative of two independent experiments ( a – i ) or combined from three independent experiments ( j–m ) or two independent experiments ( n–p ). In a separate experiment, B6 mice were i.p. inoculated with 5 × 10 5 MC38-luc cells and treated with vvDD-IL-2-RG or PBS 9 days post-tumour inoculation. Anti-CD8 Ab (250 µg per injection), anti-CD4 Ab (150 µg per injection), anti-IFN-γ Ab (200 µg per injection) (nine mice per group) ( q ), or PK136 (300 µg per injection) ( s ), were i.p. injected into mice to deplete CD8 + T cells, CD4 + T cells or neutralise circulating IFN-γ, or NK1.1 + cells, and a log-rank (Mantel-Cox) test was used to compare survival rates ( r , t ), respectively. * P

    Journal: Nature Communications

    Article Title: Modifying the cancer-immune set point using vaccinia virus expressing re-designed interleukin-2

    doi: 10.1038/s41467-018-06954-z

    Figure Lengend Snippet: Immune status change in the tumour microenvironment post-virus treatments. B6 mice were inoculated i.p. with 5 × 10 5 MC38-luc cells and treated with PBS, vvDD, vvDD-IL-2-FG, or vvDD-IL-2-RG at 2 × 10 8 PFU per mouse 9 days post-tumour inoculation. Tumour-bearing mice were sacrificed 5 days post-treatment and primary tumours were collected and analysed by flow cytometry to determine CD4 + Foxp3 - ( a ) and CD8 + IFN-γ + T cells ( b ), exhausted CD8 + T cell ( c–f ), memory-phenotype T cells (CD8 + CD44 hi ) ( g ), regulatory T cells (CD4 + Foxp3 + ) ( h ), CD8/Treg ( i ), or by RT-qPCR to determine IFN-γ, granzyme B, perforin, CXCL9, TGF-β, CD105, and VEGF ( j–p ). Four to five mice were used for each treatment group and data are representative of two independent experiments ( a – i ) or combined from three independent experiments ( j–m ) or two independent experiments ( n–p ). In a separate experiment, B6 mice were i.p. inoculated with 5 × 10 5 MC38-luc cells and treated with vvDD-IL-2-RG or PBS 9 days post-tumour inoculation. Anti-CD8 Ab (250 µg per injection), anti-CD4 Ab (150 µg per injection), anti-IFN-γ Ab (200 µg per injection) (nine mice per group) ( q ), or PK136 (300 µg per injection) ( s ), were i.p. injected into mice to deplete CD8 + T cells, CD4 + T cells or neutralise circulating IFN-γ, or NK1.1 + cells, and a log-rank (Mantel-Cox) test was used to compare survival rates ( r , t ), respectively. * P

    Article Snippet: In some experiments, anti-CD8 Ab (clone 53-6.7; Bio X Cell; 250 µg per injection), anti-CD4 Ab (clone GK1.5, Bio X Cell; 150 µg per injection), anti-NK1.1 Ab (clone PK136, Bio X Cell; 300 µg per injection), or anti-IFN-γ Ab (clone XMG1.2, Bio X Cell; 200 µg per injection) were i.p. injected into mice to deplete CD8+ T cells, CD4+ T cells, or NK1.1+ cells or neutralise circulating IFN-γ, respectively.

    Techniques: Mouse Assay, Flow Cytometry, Cytometry, Quantitative RT-PCR, Injection

    Representative Western blots (A) and bar graphs showing densitometric quantifications of the inflammatory markers interferon gamma (IFN-γ) (B) , inter-cellular adhesion molecule-1 (ICAM-1) (C) , and interleukin 17 (IL-17) (D) at 72 hours after cICH-induction

    Journal: Experimental neurology

    Article Title: Fingolimod reduces cerebral lymphocyte infiltration in experimental models of rodent intracerebral hemorrhage

    doi: 10.1016/j.expneurol.2012.12.009

    Figure Lengend Snippet: Representative Western blots (A) and bar graphs showing densitometric quantifications of the inflammatory markers interferon gamma (IFN-γ) (B) , inter-cellular adhesion molecule-1 (ICAM-1) (C) , and interleukin 17 (IL-17) (D) at 72 hours after cICH-induction

    Article Snippet: Equal amounts of protein (50 μg per sample) were separated by SDS-PAGE and transferred onto nitrocellulose membranes, which were incubated with the following primary antibodies: anti-ICAM-1, anti interferon-γ (anti-IFN-γ) (Santa Cruz Biotechnology, Santa Cruz, CA), anti interleukin-17 (anti-IL-17), and anti-β-actin (Abcam, Cambridge, MA).

    Techniques: Western Blot

    Antigen-specific whole blood assay. ( A) Time-course of IL-17A and IFN-γ production by whole blood cultured with OVA. The blood of the 10 responder cows (taken 45 days after the first immunization) was cultured in the presence of OVA (10 μg/mL) for 1 to 4 days in 96-well microplates. For each cow triplicate wells were used for each incubation time. Results are median values and quartiles (Q1; Q3). (B) The effect of magnetic depletion of CD4+ cells on the production of IL-17A and IFN-γ in the antigen-specific whole blood assay. Results obtained by stimulating blood samples from responder cows 15 days after the booster immunization are expressed as the percentage of the production by CD4+-depleted PBMC relative to cytokine production by un processed PBMC (100%). The viability of cells was not altered by the magnetic cell separation.

    Journal: PLoS ONE

    Article Title: Antigen-Specific Mammary Inflammation Depends on the Production of IL-17A and IFN-γ by Bovine CD4+ T Lymphocytes

    doi: 10.1371/journal.pone.0137755

    Figure Lengend Snippet: Antigen-specific whole blood assay. ( A) Time-course of IL-17A and IFN-γ production by whole blood cultured with OVA. The blood of the 10 responder cows (taken 45 days after the first immunization) was cultured in the presence of OVA (10 μg/mL) for 1 to 4 days in 96-well microplates. For each cow triplicate wells were used for each incubation time. Results are median values and quartiles (Q1; Q3). (B) The effect of magnetic depletion of CD4+ cells on the production of IL-17A and IFN-γ in the antigen-specific whole blood assay. Results obtained by stimulating blood samples from responder cows 15 days after the booster immunization are expressed as the percentage of the production by CD4+-depleted PBMC relative to cytokine production by un processed PBMC (100%). The viability of cells was not altered by the magnetic cell separation.

    Article Snippet: Cells were surface stained with anti-CD4 Ab, fixed, permeabilized, and stained with anti-IL-17A and anti-IFN-γ (Bio-Rad, AbD Serotec) Abs using the Cytofix/Cytoperm kit (BD Pharmingen).

    Techniques: Whole Blood Assay, Cell Culture, Incubation, Magnetic Cell Separation

    Persistence of reactivity to ovalbumin. Concentrations of IL-17A and IFN-γ yielded by the whole blood assay performed at different times after immunization with ovalbumin. Results are the median values (and interquartiles) from the 8 responder cows still available 10 months post-immunization.

    Journal: PLoS ONE

    Article Title: Antigen-Specific Mammary Inflammation Depends on the Production of IL-17A and IFN-γ by Bovine CD4+ T Lymphocytes

    doi: 10.1371/journal.pone.0137755

    Figure Lengend Snippet: Persistence of reactivity to ovalbumin. Concentrations of IL-17A and IFN-γ yielded by the whole blood assay performed at different times after immunization with ovalbumin. Results are the median values (and interquartiles) from the 8 responder cows still available 10 months post-immunization.

    Article Snippet: Cells were surface stained with anti-CD4 Ab, fixed, permeabilized, and stained with anti-IL-17A and anti-IFN-γ (Bio-Rad, AbD Serotec) Abs using the Cytofix/Cytoperm kit (BD Pharmingen).

    Techniques: Whole Blood Assay

    Intracellular expression of IL-17A and IFN-γ by CD4+ T lymphocytes. PBMC were isolated one month after ovalbumin booster injection, stimulated in vitro with ovalbumin for 3 days, rested for 2 days and finally stimulated with PMA/ionomycin for 5 h with Brefeldin A for the last 3 hours. Cells were labeled for surface CD4 and intracellular IL-17A and IFN-γ. The numbers in the plots indicate the percentages of labeled cells in comparison to the isotype control. (A) Production of viable CD4+ T lymphoblasts after culture of PBMC with (OVA) or without (NS) ovalbumin. Left panels depict PBMC from a responder cow, right panels PBMC from a low-responder. (B) PBMC from two responder cows (R1 R2) were labeled for surface CD4 and intracellular IL-17A or IFN-γ, showing CD4+ and CD4- IL-17A- and IFN-γ-producing cells. (C) labeling of CD4+ cells with anti-IL-17A and anti-IFN-γ antibodies, showing single-producing and double-producing cells. D) Double labeling of CD4+ cells from two low-responders (R3 and R4). Percentages of labeled cells are indicated in the quadrants. Results are from a representative experiment.

    Journal: PLoS ONE

    Article Title: Antigen-Specific Mammary Inflammation Depends on the Production of IL-17A and IFN-γ by Bovine CD4+ T Lymphocytes

    doi: 10.1371/journal.pone.0137755

    Figure Lengend Snippet: Intracellular expression of IL-17A and IFN-γ by CD4+ T lymphocytes. PBMC were isolated one month after ovalbumin booster injection, stimulated in vitro with ovalbumin for 3 days, rested for 2 days and finally stimulated with PMA/ionomycin for 5 h with Brefeldin A for the last 3 hours. Cells were labeled for surface CD4 and intracellular IL-17A and IFN-γ. The numbers in the plots indicate the percentages of labeled cells in comparison to the isotype control. (A) Production of viable CD4+ T lymphoblasts after culture of PBMC with (OVA) or without (NS) ovalbumin. Left panels depict PBMC from a responder cow, right panels PBMC from a low-responder. (B) PBMC from two responder cows (R1 R2) were labeled for surface CD4 and intracellular IL-17A or IFN-γ, showing CD4+ and CD4- IL-17A- and IFN-γ-producing cells. (C) labeling of CD4+ cells with anti-IL-17A and anti-IFN-γ antibodies, showing single-producing and double-producing cells. D) Double labeling of CD4+ cells from two low-responders (R3 and R4). Percentages of labeled cells are indicated in the quadrants. Results are from a representative experiment.

    Article Snippet: Cells were surface stained with anti-CD4 Ab, fixed, permeabilized, and stained with anti-IL-17A and anti-IFN-γ (Bio-Rad, AbD Serotec) Abs using the Cytofix/Cytoperm kit (BD Pharmingen).

    Techniques: Expressing, Isolation, Injection, In Vitro, Labeling

    Concentrations of cytokines in milk samples of the 10 responder cows. Time-course of concentration variation (median and interquartiles) of CXCL8 (A), IL-17A (B) and IFN- γ (C) in the milk of quarters infused with ovalbumin at 0 hpi.

    Journal: PLoS ONE

    Article Title: Antigen-Specific Mammary Inflammation Depends on the Production of IL-17A and IFN-γ by Bovine CD4+ T Lymphocytes

    doi: 10.1371/journal.pone.0137755

    Figure Lengend Snippet: Concentrations of cytokines in milk samples of the 10 responder cows. Time-course of concentration variation (median and interquartiles) of CXCL8 (A), IL-17A (B) and IFN- γ (C) in the milk of quarters infused with ovalbumin at 0 hpi.

    Article Snippet: Cells were surface stained with anti-CD4 Ab, fixed, permeabilized, and stained with anti-IL-17A and anti-IFN-γ (Bio-Rad, AbD Serotec) Abs using the Cytofix/Cytoperm kit (BD Pharmingen).

    Techniques: Concentration Assay

    Time-course of the IL-17A and IFN-γ production in the antigen-specific whole blood assay following immunization and correlation with Peak SCC. Concentrations of IL-17A (A) or IFN-γ (B) after 3 days of culture with ovalbumin of blood samples taken before and after immunization at days 0 and 30 (median values and interquartiles) distinguishing the antigen-specific responses of responders and low-responder cows to the intramammary antigenic challenge. (B and C) Correlations (Spearman’s rank test) between peak SCC and IL-17A concentrations or IFN-γ concentrations yielded by the whole blood assay performed 45 days after the first immunization.

    Journal: PLoS ONE

    Article Title: Antigen-Specific Mammary Inflammation Depends on the Production of IL-17A and IFN-γ by Bovine CD4+ T Lymphocytes

    doi: 10.1371/journal.pone.0137755

    Figure Lengend Snippet: Time-course of the IL-17A and IFN-γ production in the antigen-specific whole blood assay following immunization and correlation with Peak SCC. Concentrations of IL-17A (A) or IFN-γ (B) after 3 days of culture with ovalbumin of blood samples taken before and after immunization at days 0 and 30 (median values and interquartiles) distinguishing the antigen-specific responses of responders and low-responder cows to the intramammary antigenic challenge. (B and C) Correlations (Spearman’s rank test) between peak SCC and IL-17A concentrations or IFN-γ concentrations yielded by the whole blood assay performed 45 days after the first immunization.

    Article Snippet: Cells were surface stained with anti-CD4 Ab, fixed, permeabilized, and stained with anti-IL-17A and anti-IFN-γ (Bio-Rad, AbD Serotec) Abs using the Cytofix/Cytoperm kit (BD Pharmingen).

    Techniques: Whole Blood Assay

    Production of IFN-γ and IL-4 by AgI/II-specific cells from superficial lymph nodes (A) and spleens (B and C) of BALB/c mice immunized i.n. with AgI/II with or without CT, LT-IIa, or LT-IIb. Cells were stimulated in vitro with AgI/II at the concentrations shown for 4 days. Data shown are the arithmetic mean values ± standard deviations ( n = 4) as determined by cytokine-specific ELISA. In panels A and B, ∗, ∗∗, and ∗∗∗ indicate significant differences at P

    Journal: Infection and Immunity

    Article Title: Comparative Analysis of the Mucosal Adjuvanticity of the Type II Heat-Labile Enterotoxins LT-IIa and LT-IIb

    doi:

    Figure Lengend Snippet: Production of IFN-γ and IL-4 by AgI/II-specific cells from superficial lymph nodes (A) and spleens (B and C) of BALB/c mice immunized i.n. with AgI/II with or without CT, LT-IIa, or LT-IIb. Cells were stimulated in vitro with AgI/II at the concentrations shown for 4 days. Data shown are the arithmetic mean values ± standard deviations ( n = 4) as determined by cytokine-specific ELISA. In panels A and B, ∗, ∗∗, and ∗∗∗ indicate significant differences at P

    Article Snippet: Briefly, flat-bottomed 96-well microtiter plates (Nunc) were coated with monoclonal anti-IL-4, anti-IL-10, or anti-IFN-γ (Pharmingen) at 2 μg/ml in PBS and incubated overnight at 4°C.

    Techniques: Mouse Assay, In Vitro, Enzyme-linked Immunosorbent Assay

    ILDR2-mFc treatment inhibits myelin epitope-specific epitope spreading in the R-EAE model. ( A ) On day 76 postdisease induction, five mice from each treatment group were analyzed for recall responses to spread epitopes, via injection of 10 μg of PLP 178–191 in one ear and MBP 84–104 into the opposite ear. The level of ear swelling was assayed 24 h postchallenge. The data are presented as the mean net ear swelling. ( B ) Recall responses were also carried on splenocytes. On day 76 postdisease induction, total splenocytes were collected from five representative mice from each treatment group and activated in the presence of OVA 323–339 , PLP 139–151 , PLP 178–191 , or MBP 84–104 (20 μg/ml). Cells were pulsed with 1 mCi of tritiated thymidine at 24 h and harvested 72 h postculture set up. Cell proliferation in recall responses was also carried out using splenocytes from day-45 mice ( C ) or IFN-γ, IL-17, and IL-4 secretion was analyzed by LiquiChip following 72 h of culture ( D–F ). One representative experiment of two or three independent experiments is presented for each experimental set. * p

    Journal: Journal of immunology (Baltimore, Md. : 1950)

    Article Title: ILDR2-Fc Is a Novel Regulator of Immune Homeostasis and Inducer of Antigen-Specific Immune Tolerance

    doi: 10.4049/jimmunol.1700326

    Figure Lengend Snippet: ILDR2-mFc treatment inhibits myelin epitope-specific epitope spreading in the R-EAE model. ( A ) On day 76 postdisease induction, five mice from each treatment group were analyzed for recall responses to spread epitopes, via injection of 10 μg of PLP 178–191 in one ear and MBP 84–104 into the opposite ear. The level of ear swelling was assayed 24 h postchallenge. The data are presented as the mean net ear swelling. ( B ) Recall responses were also carried on splenocytes. On day 76 postdisease induction, total splenocytes were collected from five representative mice from each treatment group and activated in the presence of OVA 323–339 , PLP 139–151 , PLP 178–191 , or MBP 84–104 (20 μg/ml). Cells were pulsed with 1 mCi of tritiated thymidine at 24 h and harvested 72 h postculture set up. Cell proliferation in recall responses was also carried out using splenocytes from day-45 mice ( C ) or IFN-γ, IL-17, and IL-4 secretion was analyzed by LiquiChip following 72 h of culture ( D–F ). One representative experiment of two or three independent experiments is presented for each experimental set. * p

    Article Snippet: To promote differentiation, the following cytokine and Ab mixtures were added to the cultures: Th1 cells: 128 U IL-2 (National Cancer Institute), 10 ng/ml IL-12, and 10 μg/ml anti–IL-4; Th2 cells: 128 U IL-2, 10 ng/ml IL-4, 10 μg/ml IL-12, and 10 μg/ml anti–IFN-γ; and Th17 cells: 10 ng/ml TGF-β, 50 ng/ml IL-6, 4 ng/ml IL-23, 10 μg/ml anti–IL-4, and 10 μg/ml anti-IFN-γ (all from eBioscience).

    Techniques: Mouse Assay, Injection, Plasmid Purification

    ABCC3 expressed by CD56 dim CD16 + NK cells is an indicator of better patient prognosis. ( A – C ). ( A ) Representative dot plots showing that CD56 dim CD16 + NK cells express ( B ) high levels of ABCC3, ( C ) and CD56 dim CD16 + ABCC3 + NK cells express IFN-γ. ( D ) Time course of frequency of NK cells expressing ABCC3 measured by flow cytometry (* p

    Journal: International Journal of Molecular Sciences

    Article Title: ABCC3 Expressed by CD56dim CD16+ NK Cells Predicts Response in Glioblastoma Patients Treated with Combined Chemotherapy and Dendritic Cell Immunotherapy

    doi: 10.3390/ijms20235886

    Figure Lengend Snippet: ABCC3 expressed by CD56 dim CD16 + NK cells is an indicator of better patient prognosis. ( A – C ). ( A ) Representative dot plots showing that CD56 dim CD16 + NK cells express ( B ) high levels of ABCC3, ( C ) and CD56 dim CD16 + ABCC3 + NK cells express IFN-γ. ( D ) Time course of frequency of NK cells expressing ABCC3 measured by flow cytometry (* p

    Article Snippet: PBLs were then fixed and permeabilized using the Cytofix/Cytoperm solution (BD Biosciences, Franlin Lakes, NJ, USA) and intracellularly stained with an anti-IFN-γ (Miltenyi Biotec) antibody.

    Techniques: Expressing, Flow Cytometry, Cytometry

    Quantitation of soluble factors in LGL and healthy controls by ELISA and FACS. (A and C–F), Determination of expression levels of IFN-γ, IL-18/IGIF, MCP1/CCL2, IL-8/CXCL8, and IP10/CXCL10 in plasma samples from 25 to 29 LGL patients and

    Journal:

    Article Title: Phenotypic differences between healthy effector CTL and leukemic LGL cells support the notion of antigen-triggered clonal transformation in T-LGL leukemia

    doi: 10.1189/jlb.0107073

    Figure Lengend Snippet: Quantitation of soluble factors in LGL and healthy controls by ELISA and FACS. (A and C–F), Determination of expression levels of IFN-γ, IL-18/IGIF, MCP1/CCL2, IL-8/CXCL8, and IP10/CXCL10 in plasma samples from 25 to 29 LGL patients and

    Article Snippet: Finally, cells were incubated with 10 ul anti-IFN-γ (Beckman Coulter) at 4°C in the dark for 30 min. Intracellular IFN-γ production was determined within the CD3+ gate on CD8+ and CD4+ cells using a Coulter Epics XL MCL flow cytometer (Beckman Coulter).

    Techniques: Quantitation Assay, Enzyme-linked Immunosorbent Assay, FACS, Expressing

    Combined cytokine profiles in response to M. leprae . Production of IFN-γ ( A ), IP-10 ( B ) and IL-10 ( C ) determined by ELISA, in response to medium (-), PHA, M. leprae WCS or the M. leprae -unique protein ML2478 in 24 h WBA for Ethiopian leprosy patients (n = 11: 2 BT (○) and 9 BL (•), and healthy endemic controls (EC; n = 12; □). For comparison between BT and BL, significant differences were found for M. leprae WCS (Mlep) induced IFN-γ responses (p = 0.036) and ML2478 induced IL-10 responses (p = 0.035). ( D ): IP-10/IL-10 ratios are depicted for unstimulated samples after 24 h {LP (•) and EC (□)} or after 1 h WBA {LP (▵) and EC (▾)}. ( E ): Anti-PGL-I antibodies for BL (○) and BT (•) patients were detected by ELISA using natural disaccharide of PGL-I linked to HSA [31] (ND-O-HSA). Optical density (OD 450 ) readings were performed using 1∶800 serum dilutions. Median values per group are indicated by horizontal lines. The cut-off for positivity is indicated by the dashed horizontal line.

    Journal: PLoS Neglected Tropical Diseases

    Article Title: Field-Evaluation of a New Lateral Flow Assay for Detection of Cellular and Humoral Immunity against Mycobacterium leprae

    doi: 10.1371/journal.pntd.0002845

    Figure Lengend Snippet: Combined cytokine profiles in response to M. leprae . Production of IFN-γ ( A ), IP-10 ( B ) and IL-10 ( C ) determined by ELISA, in response to medium (-), PHA, M. leprae WCS or the M. leprae -unique protein ML2478 in 24 h WBA for Ethiopian leprosy patients (n = 11: 2 BT (○) and 9 BL (•), and healthy endemic controls (EC; n = 12; □). For comparison between BT and BL, significant differences were found for M. leprae WCS (Mlep) induced IFN-γ responses (p = 0.036) and ML2478 induced IL-10 responses (p = 0.035). ( D ): IP-10/IL-10 ratios are depicted for unstimulated samples after 24 h {LP (•) and EC (□)} or after 1 h WBA {LP (▵) and EC (▾)}. ( E ): Anti-PGL-I antibodies for BL (○) and BT (•) patients were detected by ELISA using natural disaccharide of PGL-I linked to HSA [31] (ND-O-HSA). Optical density (OD 450 ) readings were performed using 1∶800 serum dilutions. Median values per group are indicated by horizontal lines. The cut-off for positivity is indicated by the dashed horizontal line.

    Article Snippet: Upconverting phosphor (UCP) conjugates and LF strips UCP conjugates specific for cytokines IP-10, IL-10, IFN-γ were prepared following earlier described protocols , by conjugating 5 µg anti-IP-10 (BC-50; Diaclone), 20 µg anti-IL-10 mAb (coating mAb in ELISA, mO-13-10-12; U-CyTech) or 25 µg anti-IFN-γ (BB-1; Diaclone) per 1 mg carboxylated UCP particles, respectively.

    Techniques: Enzyme-linked Immunosorbent Assay

    Synergistic effect of macrophage and natural killer (NK) cells on hepatic apoptosis induced by lupus immunoglobulin G (IgG). (A) Western blot showing the levels of TNF-α and IFN-γ in the liver from C57BL/6 mice with intrahepatic injection of the same dose of systemic lupus erythematosus (SLE) and normal serum, SLE, and normal IgG. (B) Flow cytometry analysis of NK cell activation in the liver from B6 lpr (25 w) and age-matched control mice using anti-NK1.1, anti-CD3e and anti CD69 antibodies. The result is representative of three independent experiments, n = 6 mice per group/experiment. (C) Flow cytometry analysis of NK cell activation in the liver from C57BL/6 mice with intrahepatic injection of SLE-serum (treated for 72 h), SLE-IgG (treated for 48 h), or phosphate-buffered saline (treated 48 h) using anti-NK1.1, anti-CD3e and anti CD69 antibodies (the result is representative of three independent experiments, n = 5 mice per group/experiment). (D) Severity of liver inflammation 48 h after intrahepatic injection of SLE-IgG in C57BL/6 mice with or without NK depletion using anti-ASGM1 antibody treatment. * p

    Journal: Frontiers in Immunology

    Article Title: Role of Hepatic Deposited Immunoglobulin G in the Pathogenesis of Liver Damage in Systemic Lupus Erythematosus

    doi: 10.3389/fimmu.2018.01457

    Figure Lengend Snippet: Synergistic effect of macrophage and natural killer (NK) cells on hepatic apoptosis induced by lupus immunoglobulin G (IgG). (A) Western blot showing the levels of TNF-α and IFN-γ in the liver from C57BL/6 mice with intrahepatic injection of the same dose of systemic lupus erythematosus (SLE) and normal serum, SLE, and normal IgG. (B) Flow cytometry analysis of NK cell activation in the liver from B6 lpr (25 w) and age-matched control mice using anti-NK1.1, anti-CD3e and anti CD69 antibodies. The result is representative of three independent experiments, n = 6 mice per group/experiment. (C) Flow cytometry analysis of NK cell activation in the liver from C57BL/6 mice with intrahepatic injection of SLE-serum (treated for 72 h), SLE-IgG (treated for 48 h), or phosphate-buffered saline (treated 48 h) using anti-NK1.1, anti-CD3e and anti CD69 antibodies (the result is representative of three independent experiments, n = 5 mice per group/experiment). (D) Severity of liver inflammation 48 h after intrahepatic injection of SLE-IgG in C57BL/6 mice with or without NK depletion using anti-ASGM1 antibody treatment. * p

    Article Snippet: TNF-α and IFN-γ were detected using primary antibodies (Abs): anti-TNF-α (Abcam, ab6671) and anti-IFN-γ (Bioworld technology, BS3486) at 1/1,000 dilution.

    Techniques: Western Blot, Mouse Assay, Injection, Flow Cytometry, Cytometry, Activation Assay

    Immune checkpoint blockade induces expansion of Th1 CD4 + T cells that express T-bet and IFN-γ. ( A ) Tumor-infiltrating lymphocytes (TILs) were analyzed in day 14 tumors by flow cytometry. ICB was initiated on day 9 and repeated on day 12. Data are shown as mean ± SD from n = 9 tumors aggregated from 2 experiments. ( B ) CD4 + T cell subtype analysis in TILs by flow cytometry. ICB was initiated on day 9 and repeated on day 12. Data are shown as mean ± SD from n = 9 tumors aggregated from 2 experiments. ( C ) IFN-γ expression in intratumoral T-bet + CD4 + T cells after 4 hours of PMA/ionomycin stimulation in vitro. Data are plotted as mean ± SD from n = 5–8 tumors. ( D ) CD4 + T cells treated as in C and costained for IFN-γ and T-bet. ( E ) Similar to B , showing the percentage of T-bet + CD4 + TILs that stained for Ki67. ( F ) Percentage of Ki67 + cells in CD4 + or CD8 + T cells in dLNs on day 14 (mean ± SD). n for schematic of gating strategies. All statistical analysis by Student’s t test. NS > 0.05, *** P

    Journal: JCI Insight

    Article Title: CD4+ T cells induce rejection of urothelial tumors after immune checkpoint blockade

    doi: 10.1172/jci.insight.121062

    Figure Lengend Snippet: Immune checkpoint blockade induces expansion of Th1 CD4 + T cells that express T-bet and IFN-γ. ( A ) Tumor-infiltrating lymphocytes (TILs) were analyzed in day 14 tumors by flow cytometry. ICB was initiated on day 9 and repeated on day 12. Data are shown as mean ± SD from n = 9 tumors aggregated from 2 experiments. ( B ) CD4 + T cell subtype analysis in TILs by flow cytometry. ICB was initiated on day 9 and repeated on day 12. Data are shown as mean ± SD from n = 9 tumors aggregated from 2 experiments. ( C ) IFN-γ expression in intratumoral T-bet + CD4 + T cells after 4 hours of PMA/ionomycin stimulation in vitro. Data are plotted as mean ± SD from n = 5–8 tumors. ( D ) CD4 + T cells treated as in C and costained for IFN-γ and T-bet. ( E ) Similar to B , showing the percentage of T-bet + CD4 + TILs that stained for Ki67. ( F ) Percentage of Ki67 + cells in CD4 + or CD8 + T cells in dLNs on day 14 (mean ± SD). n for schematic of gating strategies. All statistical analysis by Student’s t test. NS > 0.05, *** P

    Article Snippet: For IFN-γ–neutralizing experiments, 250 μg/mouse isotype (PIP, Leinco Technologies) or anti–IFN-γ (H22, Leinco Technologies) antibodies were i.p. injected, as described in figure legends.

    Techniques: Flow Cytometry, Cytometry, Expressing, In Vitro, Staining

    IFN-γ mediates ICB activity and is sufficient to inhibit tumor growth. ( A ) αPD-1 and αCTLA-4 combination treatment from day 9 to 24 coadministered with IFN-γ–neutralizing antibodies administered i.p. every 3 days from day 8 to 23. Tumor sizes were compared for an additional 9 days after the last IFN-γ neutralization, a time frame within the reported half-life of the neutralizing antibody. Data represent mean tumor diameter ± SEM. n = 5 per group. ( B ) Quantification of Ck5 staining of MCB6C tumor sections obtained 5 days after initiation of combination ICB with and without IFN-γ neutralization. IFN-γ neutralization antibody was administered on days 8 and 11 after MCB6C injection. Quantification was performed using images at an original magnification of ×20. For each tumor, percentage Ck5 positivity was averaged from 4 independent fields and quantified using ImageJ software. The graph shows mean ± SD of 9 individual tumors from each treatment group. ( C ) Representative images used for B at low and high magnification. Scale bars: 1 mm (top); 200 μM (bottom). ( D ) MCB6C Infgr1-KO organoids constitutively expressing recombinant IFN-γ (rIFN-γ) were injected to mice. For all groups, mice were subjected to CD4 + ). Data are plotted as mean ± SEM of n = 6–7 mice per group. ( E ) Mass of tumors described in D at day 34. ( F ) Ck5 staining and quantification as described in B ). Comparisons for growth curves are by 2-way ANOVA for repeated measures and for column data are by Student’s t test. NS > 0.05, * P

    Journal: JCI Insight

    Article Title: CD4+ T cells induce rejection of urothelial tumors after immune checkpoint blockade

    doi: 10.1172/jci.insight.121062

    Figure Lengend Snippet: IFN-γ mediates ICB activity and is sufficient to inhibit tumor growth. ( A ) αPD-1 and αCTLA-4 combination treatment from day 9 to 24 coadministered with IFN-γ–neutralizing antibodies administered i.p. every 3 days from day 8 to 23. Tumor sizes were compared for an additional 9 days after the last IFN-γ neutralization, a time frame within the reported half-life of the neutralizing antibody. Data represent mean tumor diameter ± SEM. n = 5 per group. ( B ) Quantification of Ck5 staining of MCB6C tumor sections obtained 5 days after initiation of combination ICB with and without IFN-γ neutralization. IFN-γ neutralization antibody was administered on days 8 and 11 after MCB6C injection. Quantification was performed using images at an original magnification of ×20. For each tumor, percentage Ck5 positivity was averaged from 4 independent fields and quantified using ImageJ software. The graph shows mean ± SD of 9 individual tumors from each treatment group. ( C ) Representative images used for B at low and high magnification. Scale bars: 1 mm (top); 200 μM (bottom). ( D ) MCB6C Infgr1-KO organoids constitutively expressing recombinant IFN-γ (rIFN-γ) were injected to mice. For all groups, mice were subjected to CD4 + ). Data are plotted as mean ± SEM of n = 6–7 mice per group. ( E ) Mass of tumors described in D at day 34. ( F ) Ck5 staining and quantification as described in B ). Comparisons for growth curves are by 2-way ANOVA for repeated measures and for column data are by Student’s t test. NS > 0.05, * P

    Article Snippet: For IFN-γ–neutralizing experiments, 250 μg/mouse isotype (PIP, Leinco Technologies) or anti–IFN-γ (H22, Leinco Technologies) antibodies were i.p. injected, as described in figure legends.

    Techniques: Activity Assay, Neutralization, Staining, Injection, Software, Expressing, Recombinant, Mouse Assay

    Immune checkpoint blockade–induced rejection of MCB6C tumors is not dependent on expression of MHC I/II or Infgr1 on tumor cells. ( A ) In vivo MHC class I, MHC class II, and PD-L1 expression on MCB6C tumor cells 14 days after injection. ( B ) B2m expression, as shown by Western blot, of MCB6C using B2m-KO clones grown in vitro. ( C ) Flow cytometric evaluation of Ifngr1 expression on MCB6C Ifngr1-KO clones in vitro. ( D ) Flow cytometric evaluation of MHC II on MCB6C MHC II–KO clones in vitro, with and without IFN-γ stimulation. ( E ) Flow cytometric evaluation of MHC I on MCB6C B2m-KO clones in vitro, with and without IFN-γ stimulation. ( F ) Flow cytometric evaluation of MHC I, MHC II, and PD-L1 on MCB6C Ifngr1-KO clones in vitro, with and without IFN-γ stimulation. ( G ) In vivo tumor growth of B2m-, MHC II–, or Ifngr1-KO MCB6C lines with and without combination ICB starting 9 days after tumor injection. Data are shown as mean ± SEM. n = 5 per organoid line per treatment. ( H ) Flow cytometric evaluation of MHC I, MHC II, and PD-L1 on MCB6C Ifngr1-KO clones from MCB6C tumor cells grown in vivo and harvested on day 11.

    Journal: JCI Insight

    Article Title: CD4+ T cells induce rejection of urothelial tumors after immune checkpoint blockade

    doi: 10.1172/jci.insight.121062

    Figure Lengend Snippet: Immune checkpoint blockade–induced rejection of MCB6C tumors is not dependent on expression of MHC I/II or Infgr1 on tumor cells. ( A ) In vivo MHC class I, MHC class II, and PD-L1 expression on MCB6C tumor cells 14 days after injection. ( B ) B2m expression, as shown by Western blot, of MCB6C using B2m-KO clones grown in vitro. ( C ) Flow cytometric evaluation of Ifngr1 expression on MCB6C Ifngr1-KO clones in vitro. ( D ) Flow cytometric evaluation of MHC II on MCB6C MHC II–KO clones in vitro, with and without IFN-γ stimulation. ( E ) Flow cytometric evaluation of MHC I on MCB6C B2m-KO clones in vitro, with and without IFN-γ stimulation. ( F ) Flow cytometric evaluation of MHC I, MHC II, and PD-L1 on MCB6C Ifngr1-KO clones in vitro, with and without IFN-γ stimulation. ( G ) In vivo tumor growth of B2m-, MHC II–, or Ifngr1-KO MCB6C lines with and without combination ICB starting 9 days after tumor injection. Data are shown as mean ± SEM. n = 5 per organoid line per treatment. ( H ) Flow cytometric evaluation of MHC I, MHC II, and PD-L1 on MCB6C Ifngr1-KO clones from MCB6C tumor cells grown in vivo and harvested on day 11.

    Article Snippet: For IFN-γ–neutralizing experiments, 250 μg/mouse isotype (PIP, Leinco Technologies) or anti–IFN-γ (H22, Leinco Technologies) antibodies were i.p. injected, as described in figure legends.

    Techniques: Expressing, In Vivo, Injection, Western Blot, Clone Assay, In Vitro, Flow Cytometry

    Schematic of ICB activity in MCB6C model. Th1 CD4 + T cells increase after combination ICB and release IFN-γ, which perturbs the tumor microenvironment. ICB activity does not require MHC II expression on tumor cells, so it is presumed in this model that Th1 CD4 + T cells are stimulated within the tumor microenvironment by antigen-presenting cells that have taken up tumor antigens.

    Journal: JCI Insight

    Article Title: CD4+ T cells induce rejection of urothelial tumors after immune checkpoint blockade

    doi: 10.1172/jci.insight.121062

    Figure Lengend Snippet: Schematic of ICB activity in MCB6C model. Th1 CD4 + T cells increase after combination ICB and release IFN-γ, which perturbs the tumor microenvironment. ICB activity does not require MHC II expression on tumor cells, so it is presumed in this model that Th1 CD4 + T cells are stimulated within the tumor microenvironment by antigen-presenting cells that have taken up tumor antigens.

    Article Snippet: For IFN-γ–neutralizing experiments, 250 μg/mouse isotype (PIP, Leinco Technologies) or anti–IFN-γ (H22, Leinco Technologies) antibodies were i.p. injected, as described in figure legends.

    Techniques: Activity Assay, Expressing