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    PeproTech il 21
    Super-functional T cells in peripheral organs exceed extrafollicular T cells and Tph cells in terms of frequency. ( A ) Absolute numbers of PD-1 subsets in spleens. ( B ) Frequency of <t>IL-21</t> producers in spleens. ( C, D ) Frequency of IL-21 producers in terms of localization and in terms of PD-1 subset. Data represent two independent experiments with n = 4 mice per organ. Data are presented as the mean ± s.e.m. Figure 5A : Frequencies of PD-1 subpopulation. Data represent two independent experiments with n = 4 mice per organ. Figure 5B : Frequencies of IL-21 producers in spleens. Data represent two independent experiments with n = 4 mice per organ. Figure 5C : Frequencies of IL-21 producers in terms of localization and in terms of PD-1 subset. Data represent two independent experiments with n = 4 mice per organ. Figure 5D : Frequencies of IL-21 producers in terms of localization and in terms of PD-1 subset. Data represent two independent experiments with n = 4 mice per organ.
    Il 21, supplied by PeproTech, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/il 21/product/PeproTech
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    il 21 - by Bioz Stars, 2021-07
    86/100 stars
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    86
    r&d systems il 21
    Niclosamide ameliorates disease aggravation in R848-induced mice. Female 8-week-old C57BL/6 mice were treated with 50 μg of the TLR7 agonist R848 or control (acetone) three times weekly and were orally administered 0.5% methyl cellulose (control, R848, n = 4–6), or 100 mg/kg niclosamide (niclosamide, n = 7) daily until they were 12-week-old. a Representative photographs documenting the enlargement of spleens and cLNs. b Spleen lengths and weights in each group. c cLN lengths and weights in each group. d Urine albumin levels normalized to creatinine. e Serum levels of anti-dsDNA IgG antibody. f Serum levels of antibody subclasses (IgG, IgG2a). g Serum levels of IL-6, <t>IL-21.</t> h Left, representative photomicrographs of PAS-stained sections of kidney. Original magnification ×200 (upper), ×100 (bottom). Right, histological scores. i Left, representative immunofluorescent images of kidney C3 staining. Original magnification ×100 (upper), ×400 (bottom). Right, MFI of C3 deposition. Data shown as mean ± SD. One-way ANOVA was performed. * P
    Il 21, supplied by r&d systems, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/il 21/product/r&d systems
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    il 21 - by Bioz Stars, 2021-07
    86/100 stars
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    94
    R&D Systems rmil 21
    Niclosamide ameliorates disease aggravation in R848-induced mice. Female 8-week-old C57BL/6 mice were treated with 50 μg of the TLR7 agonist R848 or control (acetone) three times weekly and were orally administered 0.5% methyl cellulose (control, R848, n = 4–6), or 100 mg/kg niclosamide (niclosamide, n = 7) daily until they were 12-week-old. a Representative photographs documenting the enlargement of spleens and cLNs. b Spleen lengths and weights in each group. c cLN lengths and weights in each group. d Urine albumin levels normalized to creatinine. e Serum levels of anti-dsDNA IgG antibody. f Serum levels of antibody subclasses (IgG, IgG2a). g Serum levels of IL-6, <t>IL-21.</t> h Left, representative photomicrographs of PAS-stained sections of kidney. Original magnification ×200 (upper), ×100 (bottom). Right, histological scores. i Left, representative immunofluorescent images of kidney C3 staining. Original magnification ×100 (upper), ×400 (bottom). Right, MFI of C3 deposition. Data shown as mean ± SD. One-way ANOVA was performed. * P
    Rmil 21, supplied by R&D Systems, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rmil 21/product/R&D Systems
    Average 94 stars, based on 1 article reviews
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    rmil 21 - by Bioz Stars, 2021-07
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    94
    Thermo Fisher gene exp il21 mm00517640 m1
    Potent Tfh cell responses are elicited by a single immunization with m1Ψ-mRNA-LNPs in mice. Mice were immunized once i.d. with 30 µg of Luc or HA m1Ψ-mRNA-LNPs, a single i.m. injection with 1,000 HAU of inactivated PR8 virus, MF59-adjuvanted recombinant PR8 HA protein, or intranasally infected with 25 TCID 50 of live PR8 influenza virus, and immune responses were examined 10 d after immunization (A and B). (A) Total numbers of splenic Tfh cells were determined by staining for TCR + CD19 − CD4 + CD62L − CXCR5 + PD-1 + T cells. (B) IFN-γ, IL-4, and <t>IL-21</t> transcript levels in sorted Tfh cells from PR8 HA m1Ψ-mRNA-LNP–immunized mice were determined by quantitative real-time RT-PCR. Fold induction of cytokines compared with total universal RNA is shown. (C and D) Mice were immunized with a single i.d. injection of 30 µg of HA or Env m1Ψ-mRNA-LNPs, and immune responses were examined 12 d after immunization. Percentage of IFN-γ producing CD4 + Bcl-6 + Tfh-like cells was measured by flow cytometry after HA (C) or Env (D) peptide stimulation. (E) Mice were immunized with a single i.d. injection of 30 µg of PR8 HA m1Ψ-mRNA-LNPs, and rates of binding to PR8 HA were examined 2, 4, and 8 wk later by biolayer interferometry. The apparent nanomolar affinity of anti–HA antibodies, derived from the mean rates of HA-binding in polyclonal sera, is plotted for each serum sample, with lower values corresponding to higher apparent affinity. n = 5–8 mice, and each symbol represents values for one animal. Experiments were repeated at least two times to achieve sufficient numbers of values for mice in each group. Error bars are SEM. Statistical analysis: one-way ANOVA with Bonferroni correction, *, P
    Gene Exp Il21 Mm00517640 M1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 94 stars, based on 1 article reviews
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    gene exp il21 mm00517640 m1 - by Bioz Stars, 2021-07
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    N/A
    Source Escherichia Coli Interleukin 21 Mouse Recombinant produced in E Coli is a single non glycosilated polypeptide chain containing 130 amino acids and having a total molecular mass of 15kDa
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    N/A
    RayBio Mouse Immunoquantitative PCR Based IL 21 ELISA Kit for cell culture supernatants plasma and serum samples
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    N/A
    The Mouse IL 21 Antibody from R D Systems is a rat monoclonal antibody to IL 21 This antibody reacts with mouse The Mouse IL 21 Antibody has been validated
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    Image Search Results


    Super-functional T cells in peripheral organs exceed extrafollicular T cells and Tph cells in terms of frequency. ( A ) Absolute numbers of PD-1 subsets in spleens. ( B ) Frequency of IL-21 producers in spleens. ( C, D ) Frequency of IL-21 producers in terms of localization and in terms of PD-1 subset. Data represent two independent experiments with n = 4 mice per organ. Data are presented as the mean ± s.e.m. Figure 5A : Frequencies of PD-1 subpopulation. Data represent two independent experiments with n = 4 mice per organ. Figure 5B : Frequencies of IL-21 producers in spleens. Data represent two independent experiments with n = 4 mice per organ. Figure 5C : Frequencies of IL-21 producers in terms of localization and in terms of PD-1 subset. Data represent two independent experiments with n = 4 mice per organ. Figure 5D : Frequencies of IL-21 producers in terms of localization and in terms of PD-1 subset. Data represent two independent experiments with n = 4 mice per organ.

    Journal: eLife

    Article Title: Identification of a super-functional Tfh-like subpopulation in murine lupus by pattern perception

    doi: 10.7554/eLife.53226

    Figure Lengend Snippet: Super-functional T cells in peripheral organs exceed extrafollicular T cells and Tph cells in terms of frequency. ( A ) Absolute numbers of PD-1 subsets in spleens. ( B ) Frequency of IL-21 producers in spleens. ( C, D ) Frequency of IL-21 producers in terms of localization and in terms of PD-1 subset. Data represent two independent experiments with n = 4 mice per organ. Data are presented as the mean ± s.e.m. Figure 5A : Frequencies of PD-1 subpopulation. Data represent two independent experiments with n = 4 mice per organ. Figure 5B : Frequencies of IL-21 producers in spleens. Data represent two independent experiments with n = 4 mice per organ. Figure 5C : Frequencies of IL-21 producers in terms of localization and in terms of PD-1 subset. Data represent two independent experiments with n = 4 mice per organ. Figure 5D : Frequencies of IL-21 producers in terms of localization and in terms of PD-1 subset. Data represent two independent experiments with n = 4 mice per organ.

    Article Snippet: However, PRI assessment of the same samples (Figure 3F) provides a visual representation suggesting that the vast majority of IFNγ+ cells are also producing IL-21, while in the dot plots (Figure 3A) only ~1/7 of IFNγ+ cells produce IL-21.

    Techniques: Functional Assay, Mouse Assay

    Functional comparison of CXCR3 + PD-1 lo Tsh, CXCR3 - PD-1 lo CD4 + T cells and PD-1 hi cells in B and T cell co-cultures. ( A ) Gating strategy used to sort CXCR3 + PD-1 lo Tsh, CXCR3 - PD-1 lo CD4 + T cells and PD-1 hi CXCR5 +/- . Upper row (pseudo-color plots) shows pre-sorted cells prepared from pooled splenocytes of two-years old C57Bl/6 mice. Lower row (black dot plots) shows purity and phenotype of the sorted populations. ( B ) Gating strategy to sort B220 + CD19 + B cells. Pseudo-color plots on the left-hand side show pre-sorted cells prepared from pooled splenocytes of 2-year-old C57Bl/6 mice and enriched for B cells by negative magnetic cell sort on Miltenyi column. Black dot plots on the right-hand side show B220 + CD19 + B cells purity after FACS-sort. ( C ) PMA/ionomycin-stimulated pre-sorted cells from pooled splenocytes of two-years old C57Bl/6 mice were assessed for CD40L, IL-21, and IFN-γ production. Pseudo-color plots show gating strategy for identification of CD4 T cell subsets and contour plots show cytokine production by the assessed populations. ( D ) Analysis of 5 days co-cultures by flow cytometry. Representative plots from co-culture wells with B cells and PD-1 hi cells (upper plots) and B cells and CXCR3 + PD-1 lo Tsh cells (lower plots) are shown. Data are representative of two independent experiments. Legends for figure source data.

    Journal: eLife

    Article Title: Identification of a super-functional Tfh-like subpopulation in murine lupus by pattern perception

    doi: 10.7554/eLife.53226

    Figure Lengend Snippet: Functional comparison of CXCR3 + PD-1 lo Tsh, CXCR3 - PD-1 lo CD4 + T cells and PD-1 hi cells in B and T cell co-cultures. ( A ) Gating strategy used to sort CXCR3 + PD-1 lo Tsh, CXCR3 - PD-1 lo CD4 + T cells and PD-1 hi CXCR5 +/- . Upper row (pseudo-color plots) shows pre-sorted cells prepared from pooled splenocytes of two-years old C57Bl/6 mice. Lower row (black dot plots) shows purity and phenotype of the sorted populations. ( B ) Gating strategy to sort B220 + CD19 + B cells. Pseudo-color plots on the left-hand side show pre-sorted cells prepared from pooled splenocytes of 2-year-old C57Bl/6 mice and enriched for B cells by negative magnetic cell sort on Miltenyi column. Black dot plots on the right-hand side show B220 + CD19 + B cells purity after FACS-sort. ( C ) PMA/ionomycin-stimulated pre-sorted cells from pooled splenocytes of two-years old C57Bl/6 mice were assessed for CD40L, IL-21, and IFN-γ production. Pseudo-color plots show gating strategy for identification of CD4 T cell subsets and contour plots show cytokine production by the assessed populations. ( D ) Analysis of 5 days co-cultures by flow cytometry. Representative plots from co-culture wells with B cells and PD-1 hi cells (upper plots) and B cells and CXCR3 + PD-1 lo Tsh cells (lower plots) are shown. Data are representative of two independent experiments. Legends for figure source data.

    Article Snippet: However, PRI assessment of the same samples (Figure 3F) provides a visual representation suggesting that the vast majority of IFNγ+ cells are also producing IL-21, while in the dot plots (Figure 3A) only ~1/7 of IFNγ+ cells produce IL-21.

    Techniques: Functional Assay, Mouse Assay, FACS, Flow Cytometry, Co-Culture Assay

    PRI results can be confirmed with viSNE and conventional analysis. ( A ) Bar plots of subpopulation frequencies sub-divided into regions as described in Figure 6 ( A, D ). ( B ) viSNE plots displaying cell density and MFI of different markers. Grey circles mark the PD-1hi area and red circles the IFN-γ hi area. ( C ) Bin plots displaying PD-1 (x-axis) and IL-21 (y-axis) with cell density and MFI+ of IFN-γ, Bcl6, CXCR5 and ICOS. Cell frequencies per quadrant are calculated on the number of cells per sample (black), number of Z + cells per sample (green), and number of Z + cells per quadrant to all Z+ cells (blue). Grey bins contain less than 10 Z + cells. ( D ) FlowJo color map with PD-1 (x-axis), IFN-γ (y-axis) and MFI of IL-21 (Z parameter). ( E ) 3D heatmap plots showing MFI+ of Bcl6 (left) and IL-21 (right) as relief on PD-1 (x-axis) and IFN-γ (y-axis). Data are representative for at least two independent experiments with old diseased mice with ( A ) n = 3–11 mice and ( B–E ) n ≥ 3 mice. A, Data are presented as the mean ± s.e.m. Figure 6—figure supplement 1A : Frequencies of protein expressions sub-divided into regions. Data represent three independent experiments with n = 3–11 mice.

    Journal: eLife

    Article Title: Identification of a super-functional Tfh-like subpopulation in murine lupus by pattern perception

    doi: 10.7554/eLife.53226

    Figure Lengend Snippet: PRI results can be confirmed with viSNE and conventional analysis. ( A ) Bar plots of subpopulation frequencies sub-divided into regions as described in Figure 6 ( A, D ). ( B ) viSNE plots displaying cell density and MFI of different markers. Grey circles mark the PD-1hi area and red circles the IFN-γ hi area. ( C ) Bin plots displaying PD-1 (x-axis) and IL-21 (y-axis) with cell density and MFI+ of IFN-γ, Bcl6, CXCR5 and ICOS. Cell frequencies per quadrant are calculated on the number of cells per sample (black), number of Z + cells per sample (green), and number of Z + cells per quadrant to all Z+ cells (blue). Grey bins contain less than 10 Z + cells. ( D ) FlowJo color map with PD-1 (x-axis), IFN-γ (y-axis) and MFI of IL-21 (Z parameter). ( E ) 3D heatmap plots showing MFI+ of Bcl6 (left) and IL-21 (right) as relief on PD-1 (x-axis) and IFN-γ (y-axis). Data are representative for at least two independent experiments with old diseased mice with ( A ) n = 3–11 mice and ( B–E ) n ≥ 3 mice. A, Data are presented as the mean ± s.e.m. Figure 6—figure supplement 1A : Frequencies of protein expressions sub-divided into regions. Data represent three independent experiments with n = 3–11 mice.

    Article Snippet: However, PRI assessment of the same samples (Figure 3F) provides a visual representation suggesting that the vast majority of IFNγ+ cells are also producing IL-21, while in the dot plots (Figure 3A) only ~1/7 of IFNγ+ cells produce IL-21.

    Techniques: Mouse Assay

    The majority of IL-21 is produced by non-Tfh cells. ( A ) Co-production of IFN-γ, IL-2, IL-10, IL-21 and TNF-α was analyzed by a pie chart. ( B, C ) PRI-based statistical analysis of marker co-expression in young and old mice. ( D ) Bin plots of PD-1 (x-axis) vs. CXCR5 (y-axis) with heatmap of frequency (top) and expression level (bottom) per bin of Tfh and B cell interaction proteins. Cell frequencies per quadrant are calculated on the number of cells per sample (black) and number of Z + cells per sample (green). Grey bins contain less than 10 Z + cells. ( D ) Data represent two experiments with n = 6 mice in total. ( B, C ) Samples were compared using the unpaired two-tailed t-test. Data are presented as the mean ± s.e.m. Figure 3—figure supplement 1A : Raw data to determine the frequencies of boolean combinations of coexpression of five cytokines. Figure 3—figure supplement 1B, C : Frequencies from IL-21 + subpopulations extracted from PRI bin plots. Data as in Figure 3—source data 1 .

    Journal: eLife

    Article Title: Identification of a super-functional Tfh-like subpopulation in murine lupus by pattern perception

    doi: 10.7554/eLife.53226

    Figure Lengend Snippet: The majority of IL-21 is produced by non-Tfh cells. ( A ) Co-production of IFN-γ, IL-2, IL-10, IL-21 and TNF-α was analyzed by a pie chart. ( B, C ) PRI-based statistical analysis of marker co-expression in young and old mice. ( D ) Bin plots of PD-1 (x-axis) vs. CXCR5 (y-axis) with heatmap of frequency (top) and expression level (bottom) per bin of Tfh and B cell interaction proteins. Cell frequencies per quadrant are calculated on the number of cells per sample (black) and number of Z + cells per sample (green). Grey bins contain less than 10 Z + cells. ( D ) Data represent two experiments with n = 6 mice in total. ( B, C ) Samples were compared using the unpaired two-tailed t-test. Data are presented as the mean ± s.e.m. Figure 3—figure supplement 1A : Raw data to determine the frequencies of boolean combinations of coexpression of five cytokines. Figure 3—figure supplement 1B, C : Frequencies from IL-21 + subpopulations extracted from PRI bin plots. Data as in Figure 3—source data 1 .

    Article Snippet: However, PRI assessment of the same samples (Figure 3F) provides a visual representation suggesting that the vast majority of IFNγ+ cells are also producing IL-21, while in the dot plots (Figure 3A) only ~1/7 of IFNγ+ cells produce IL-21.

    Techniques: Produced, Marker, Expressing, Mouse Assay, Two Tailed Test

    Niclosamide ameliorates disease aggravation in R848-induced mice. Female 8-week-old C57BL/6 mice were treated with 50 μg of the TLR7 agonist R848 or control (acetone) three times weekly and were orally administered 0.5% methyl cellulose (control, R848, n = 4–6), or 100 mg/kg niclosamide (niclosamide, n = 7) daily until they were 12-week-old. a Representative photographs documenting the enlargement of spleens and cLNs. b Spleen lengths and weights in each group. c cLN lengths and weights in each group. d Urine albumin levels normalized to creatinine. e Serum levels of anti-dsDNA IgG antibody. f Serum levels of antibody subclasses (IgG, IgG2a). g Serum levels of IL-6, IL-21. h Left, representative photomicrographs of PAS-stained sections of kidney. Original magnification ×200 (upper), ×100 (bottom). Right, histological scores. i Left, representative immunofluorescent images of kidney C3 staining. Original magnification ×100 (upper), ×400 (bottom). Right, MFI of C3 deposition. Data shown as mean ± SD. One-way ANOVA was performed. * P

    Journal: Journal of Translational Medicine

    Article Title: Niclosamide suppresses the expansion of follicular helper T cells and alleviates disease severity in two murine models of lupus via STAT3

    doi: 10.1186/s12967-021-02760-2

    Figure Lengend Snippet: Niclosamide ameliorates disease aggravation in R848-induced mice. Female 8-week-old C57BL/6 mice were treated with 50 μg of the TLR7 agonist R848 or control (acetone) three times weekly and were orally administered 0.5% methyl cellulose (control, R848, n = 4–6), or 100 mg/kg niclosamide (niclosamide, n = 7) daily until they were 12-week-old. a Representative photographs documenting the enlargement of spleens and cLNs. b Spleen lengths and weights in each group. c cLN lengths and weights in each group. d Urine albumin levels normalized to creatinine. e Serum levels of anti-dsDNA IgG antibody. f Serum levels of antibody subclasses (IgG, IgG2a). g Serum levels of IL-6, IL-21. h Left, representative photomicrographs of PAS-stained sections of kidney. Original magnification ×200 (upper), ×100 (bottom). Right, histological scores. i Left, representative immunofluorescent images of kidney C3 staining. Original magnification ×100 (upper), ×400 (bottom). Right, MFI of C3 deposition. Data shown as mean ± SD. One-way ANOVA was performed. * P

    Article Snippet: For TFH -like cell differentiation, purified CD4+ T cells were seeded at 1 × 106 cells/well and were activated with mouse T-activator CD3/CD28 Dynabeads™ (Invitrogen), and treated with 20 ng/ml IL-6, 20 ng/ml IL-21, 10 μg/ml anti-IL-4, 10 μg/ml anti-IFN-γ, and 20 μg/ml anti-TGF-β (R & D Systems) for 4 days with or without niclosamide.

    Techniques: Mouse Assay, Staining

    Niclosamide ameliorates disease aggravation in MRL/ lpr mice. Female 10-week-old MRL/ lpr mice were orally administered 0.5% methyl cellulose (vehicle, n = 7), or 100 mg/kg niclosamide (Niclosamide, n = 7) daily until they were 16-week-old. a Left, Representative photographs documenting the enlargement of kidneys. Right, kidney weights in vehicle, niclosamide groups. b Urine albumin levels normalized to creatinine. c Serum levels of anti-dsDNA IgG antibody. d Serum levels of antibody subclasses (IgG, IgG1, and IgM). e Serum levels of IL-6 and IL-21. f Left, representative photomicrographs of PAS-stained sections of kidney. Original magnification ×200 (upper), ×100 (middle, bottom). Right, histological score. g Left, representative immunofluorescent images of kidney C3 staining. Original magnification ×100 (upper), ×400 (bottom). Right, MFI of C3 deposition. Data shown as mean ± SD. t-test was performed. * P

    Journal: Journal of Translational Medicine

    Article Title: Niclosamide suppresses the expansion of follicular helper T cells and alleviates disease severity in two murine models of lupus via STAT3

    doi: 10.1186/s12967-021-02760-2

    Figure Lengend Snippet: Niclosamide ameliorates disease aggravation in MRL/ lpr mice. Female 10-week-old MRL/ lpr mice were orally administered 0.5% methyl cellulose (vehicle, n = 7), or 100 mg/kg niclosamide (Niclosamide, n = 7) daily until they were 16-week-old. a Left, Representative photographs documenting the enlargement of kidneys. Right, kidney weights in vehicle, niclosamide groups. b Urine albumin levels normalized to creatinine. c Serum levels of anti-dsDNA IgG antibody. d Serum levels of antibody subclasses (IgG, IgG1, and IgM). e Serum levels of IL-6 and IL-21. f Left, representative photomicrographs of PAS-stained sections of kidney. Original magnification ×200 (upper), ×100 (middle, bottom). Right, histological score. g Left, representative immunofluorescent images of kidney C3 staining. Original magnification ×100 (upper), ×400 (bottom). Right, MFI of C3 deposition. Data shown as mean ± SD. t-test was performed. * P

    Article Snippet: For TFH -like cell differentiation, purified CD4+ T cells were seeded at 1 × 106 cells/well and were activated with mouse T-activator CD3/CD28 Dynabeads™ (Invitrogen), and treated with 20 ng/ml IL-6, 20 ng/ml IL-21, 10 μg/ml anti-IL-4, 10 μg/ml anti-IFN-γ, and 20 μg/ml anti-TGF-β (R & D Systems) for 4 days with or without niclosamide.

    Techniques: Mouse Assay, Staining

    Niclosamide inhibits T FH -like cell differentiation and B cell IgG production in vitro. Naive CD4 + T cells were purified from the spleens of MRL/ lpr and C57BL/6 mice. For T FH -like cell differentiation, purified CD4 + T cells were activated with mouse T-activator CD3/CD28 Dynabeads, and treated with 20 ng/ml IL-6, 20 ng/ml IL-21, 10 μg/ml anti-IL-4, 10 μg/ml anti-IFN-γ, and 20 μg/ml anti-TGF-β for 4 days with or without niclosamide. a Left, T FH -like cells (CXCR5 + PD-1 + , gated on CD4 + ) isolated from MRL/ lpr mice were analyzed by flow cytometry. Right, the percentage of T FH -like cells is shown. b p-STAT3 and TCF-1 expressions in T FH -like cells isolated from MRL/ lpr mice were analyzed by western blot. c mRNA expression levels of Bcl-6 , CXCR5 , and Blimp-1 in T FH -like cells isolated from MRL/ lpr mice were measured by real-time PCR. d T FH -like cells and B cells isolated from C57BL/6 mice were co-cultured with or without T FH -like cells for 3 days, and then the concentrations of IgG in the supernatants were detected by ELISA. Data shown as mean ± SD. One-way ANOVA was performed. ** P

    Journal: Journal of Translational Medicine

    Article Title: Niclosamide suppresses the expansion of follicular helper T cells and alleviates disease severity in two murine models of lupus via STAT3

    doi: 10.1186/s12967-021-02760-2

    Figure Lengend Snippet: Niclosamide inhibits T FH -like cell differentiation and B cell IgG production in vitro. Naive CD4 + T cells were purified from the spleens of MRL/ lpr and C57BL/6 mice. For T FH -like cell differentiation, purified CD4 + T cells were activated with mouse T-activator CD3/CD28 Dynabeads, and treated with 20 ng/ml IL-6, 20 ng/ml IL-21, 10 μg/ml anti-IL-4, 10 μg/ml anti-IFN-γ, and 20 μg/ml anti-TGF-β for 4 days with or without niclosamide. a Left, T FH -like cells (CXCR5 + PD-1 + , gated on CD4 + ) isolated from MRL/ lpr mice were analyzed by flow cytometry. Right, the percentage of T FH -like cells is shown. b p-STAT3 and TCF-1 expressions in T FH -like cells isolated from MRL/ lpr mice were analyzed by western blot. c mRNA expression levels of Bcl-6 , CXCR5 , and Blimp-1 in T FH -like cells isolated from MRL/ lpr mice were measured by real-time PCR. d T FH -like cells and B cells isolated from C57BL/6 mice were co-cultured with or without T FH -like cells for 3 days, and then the concentrations of IgG in the supernatants were detected by ELISA. Data shown as mean ± SD. One-way ANOVA was performed. ** P

    Article Snippet: For TFH -like cell differentiation, purified CD4+ T cells were seeded at 1 × 106 cells/well and were activated with mouse T-activator CD3/CD28 Dynabeads™ (Invitrogen), and treated with 20 ng/ml IL-6, 20 ng/ml IL-21, 10 μg/ml anti-IL-4, 10 μg/ml anti-IFN-γ, and 20 μg/ml anti-TGF-β (R & D Systems) for 4 days with or without niclosamide.

    Techniques: Cell Differentiation, In Vitro, Purification, Mouse Assay, Isolation, Flow Cytometry, Western Blot, Expressing, Real-time Polymerase Chain Reaction, Cell Culture, Enzyme-linked Immunosorbent Assay

    Niclosamide treatment reduces T FH -associated gene and cytokine levels in the spleens of MRL/ lpr mice. a p-STAT3, Bcl-6, and TCF-1 expression in the spleen were analyzed by western blot. b mRNA expression levels of Bcl-6 , CXCR5 , and Blimp-1 in CD4 + T cells purified from spleens for each group of mice were measured by real-time PCR. c The levels of IL-6 and IL-21 in the spleen homogenates were detected by ELISA. Data shown as mean ± SD. t-test was performed. ** P

    Journal: Journal of Translational Medicine

    Article Title: Niclosamide suppresses the expansion of follicular helper T cells and alleviates disease severity in two murine models of lupus via STAT3

    doi: 10.1186/s12967-021-02760-2

    Figure Lengend Snippet: Niclosamide treatment reduces T FH -associated gene and cytokine levels in the spleens of MRL/ lpr mice. a p-STAT3, Bcl-6, and TCF-1 expression in the spleen were analyzed by western blot. b mRNA expression levels of Bcl-6 , CXCR5 , and Blimp-1 in CD4 + T cells purified from spleens for each group of mice were measured by real-time PCR. c The levels of IL-6 and IL-21 in the spleen homogenates were detected by ELISA. Data shown as mean ± SD. t-test was performed. ** P

    Article Snippet: For TFH -like cell differentiation, purified CD4+ T cells were seeded at 1 × 106 cells/well and were activated with mouse T-activator CD3/CD28 Dynabeads™ (Invitrogen), and treated with 20 ng/ml IL-6, 20 ng/ml IL-21, 10 μg/ml anti-IL-4, 10 μg/ml anti-IFN-γ, and 20 μg/ml anti-TGF-β (R & D Systems) for 4 days with or without niclosamide.

    Techniques: Mouse Assay, Expressing, Western Blot, Purification, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

    Potent Tfh cell responses are elicited by a single immunization with m1Ψ-mRNA-LNPs in mice. Mice were immunized once i.d. with 30 µg of Luc or HA m1Ψ-mRNA-LNPs, a single i.m. injection with 1,000 HAU of inactivated PR8 virus, MF59-adjuvanted recombinant PR8 HA protein, or intranasally infected with 25 TCID 50 of live PR8 influenza virus, and immune responses were examined 10 d after immunization (A and B). (A) Total numbers of splenic Tfh cells were determined by staining for TCR + CD19 − CD4 + CD62L − CXCR5 + PD-1 + T cells. (B) IFN-γ, IL-4, and IL-21 transcript levels in sorted Tfh cells from PR8 HA m1Ψ-mRNA-LNP–immunized mice were determined by quantitative real-time RT-PCR. Fold induction of cytokines compared with total universal RNA is shown. (C and D) Mice were immunized with a single i.d. injection of 30 µg of HA or Env m1Ψ-mRNA-LNPs, and immune responses were examined 12 d after immunization. Percentage of IFN-γ producing CD4 + Bcl-6 + Tfh-like cells was measured by flow cytometry after HA (C) or Env (D) peptide stimulation. (E) Mice were immunized with a single i.d. injection of 30 µg of PR8 HA m1Ψ-mRNA-LNPs, and rates of binding to PR8 HA were examined 2, 4, and 8 wk later by biolayer interferometry. The apparent nanomolar affinity of anti–HA antibodies, derived from the mean rates of HA-binding in polyclonal sera, is plotted for each serum sample, with lower values corresponding to higher apparent affinity. n = 5–8 mice, and each symbol represents values for one animal. Experiments were repeated at least two times to achieve sufficient numbers of values for mice in each group. Error bars are SEM. Statistical analysis: one-way ANOVA with Bonferroni correction, *, P

    Journal: The Journal of Experimental Medicine

    Article Title: Nucleoside-modified mRNA vaccines induce potent T follicular helper and germinal center B cell responses

    doi: 10.1084/jem.20171450

    Figure Lengend Snippet: Potent Tfh cell responses are elicited by a single immunization with m1Ψ-mRNA-LNPs in mice. Mice were immunized once i.d. with 30 µg of Luc or HA m1Ψ-mRNA-LNPs, a single i.m. injection with 1,000 HAU of inactivated PR8 virus, MF59-adjuvanted recombinant PR8 HA protein, or intranasally infected with 25 TCID 50 of live PR8 influenza virus, and immune responses were examined 10 d after immunization (A and B). (A) Total numbers of splenic Tfh cells were determined by staining for TCR + CD19 − CD4 + CD62L − CXCR5 + PD-1 + T cells. (B) IFN-γ, IL-4, and IL-21 transcript levels in sorted Tfh cells from PR8 HA m1Ψ-mRNA-LNP–immunized mice were determined by quantitative real-time RT-PCR. Fold induction of cytokines compared with total universal RNA is shown. (C and D) Mice were immunized with a single i.d. injection of 30 µg of HA or Env m1Ψ-mRNA-LNPs, and immune responses were examined 12 d after immunization. Percentage of IFN-γ producing CD4 + Bcl-6 + Tfh-like cells was measured by flow cytometry after HA (C) or Env (D) peptide stimulation. (E) Mice were immunized with a single i.d. injection of 30 µg of PR8 HA m1Ψ-mRNA-LNPs, and rates of binding to PR8 HA were examined 2, 4, and 8 wk later by biolayer interferometry. The apparent nanomolar affinity of anti–HA antibodies, derived from the mean rates of HA-binding in polyclonal sera, is plotted for each serum sample, with lower values corresponding to higher apparent affinity. n = 5–8 mice, and each symbol represents values for one animal. Experiments were repeated at least two times to achieve sufficient numbers of values for mice in each group. Error bars are SEM. Statistical analysis: one-way ANOVA with Bonferroni correction, *, P

    Article Snippet: Probes used were Il4 (Mm00445260_m1), Ifnγ (Mm00801778_m1), Il21 (Mm00517640_m1), and Gapdh (Mm99999915_g1).

    Techniques: Mouse Assay, Injection, Recombinant, Infection, Staining, Quantitative RT-PCR, Flow Cytometry, Cytometry, Binding Assay, Derivative Assay