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human th2 cell differentiation kit  (R&D Systems)


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    R&D Systems human th2 cell differentiation kit
    Human Th2 Cell Differentiation Kit, supplied by R&D Systems, used in various techniques. Bioz Stars score: 94/100, based on 25 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/cellxvivo+human+th2+differentiation+kit/CellXVivo+Human+Treg+Cell+Differentiation+Kit/pm39067169-42-25-30
    Average 94 stars, based on 25 article reviews
    human th2 cell differentiation kit - by Bioz Stars, 2026-09
    94/100 stars

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    Related Articles

    Enzyme-linked Immunosorbent Assay:

    Article Title: The ammonia oxidizing bacterium Nitrosomonas eutropha blocks T helper 2 cell polarization via the anti-inflammatory cytokine IL-10
    Article Snippet: AOB were added at a ratio of four bacteria per PBMC 1 h prior to Th2 stimulation.AOB were added at a ratio of four bacteria per PBMC 1 h prior to Th2 stimulation.. PBMC were then stimulated using either the CellXvivo human Th2 differentiation kit (R&D systems) following the manufacturer’s recommendations, or 1 µg/mL SEB (Sigma).. Where appropriate, neutralizing antibodies were used at 10 µg/mL and refreshed daily until sample collection.Where appropriate, neutralizing antibodies were used at 10 µg/mL and refreshed daily until sample collection.

    Article Title: The ammonia oxidizing bacterium Nitrosomonas eutropha D23 blocks T helper 2 cell polarization via IL-10 – mediated interference with dendritic cell activation
    Article Snippet: AOB were added at a ratio of four bacteria per PBMC one hour prior to Th2 stimulation.AOB were added at a ratio of four bacteria per PBMC one hour prior to Th2 stimulation.. PBMC were then stimulated using either the CellXvivo human Th2 differentiation kit (R&D systems) following the manufacturer’s recommendations, or 1μg/mL SEB (Sigma).. Where appropriate, neutralizing antibodies were used at 10μg/mL and refreshed daily until sample collection.Where appropriate, neutralizing antibodies were used at 10μg/mL and refreshed daily until sample collection.

    Gene Expression:

    Article Title: The ammonia oxidizing bacterium Nitrosomonas eutropha blocks T helper 2 cell polarization via the anti-inflammatory cytokine IL-10
    Article Snippet: AOB were added at a ratio of four bacteria per PBMC 1 h prior to Th2 stimulation.AOB were added at a ratio of four bacteria per PBMC 1 h prior to Th2 stimulation.. PBMC were then stimulated using either the CellXvivo human Th2 differentiation kit (R&D systems) following the manufacturer’s recommendations, or 1 µg/mL SEB (Sigma).. Where appropriate, neutralizing antibodies were used at 10 µg/mL and refreshed daily until sample collection.Where appropriate, neutralizing antibodies were used at 10 µg/mL and refreshed daily until sample collection.

    Article Title: The ammonia oxidizing bacterium Nitrosomonas eutropha D23 blocks T helper 2 cell polarization via IL-10 – mediated interference with dendritic cell activation
    Article Snippet: AOB were added at a ratio of four bacteria per PBMC one hour prior to Th2 stimulation.AOB were added at a ratio of four bacteria per PBMC one hour prior to Th2 stimulation.. PBMC were then stimulated using either the CellXvivo human Th2 differentiation kit (R&D systems) following the manufacturer’s recommendations, or 1μg/mL SEB (Sigma).. Where appropriate, neutralizing antibodies were used at 10μg/mL and refreshed daily until sample collection.Where appropriate, neutralizing antibodies were used at 10μg/mL and refreshed daily until sample collection.

    Expressing:

    Article Title: The ammonia oxidizing bacterium Nitrosomonas eutropha blocks T helper 2 cell polarization via the anti-inflammatory cytokine IL-10
    Article Snippet: AOB were added at a ratio of four bacteria per PBMC 1 h prior to Th2 stimulation.AOB were added at a ratio of four bacteria per PBMC 1 h prior to Th2 stimulation.. PBMC were then stimulated using either the CellXvivo human Th2 differentiation kit (R&D systems) following the manufacturer’s recommendations, or 1 µg/mL SEB (Sigma).. Where appropriate, neutralizing antibodies were used at 10 µg/mL and refreshed daily until sample collection.Where appropriate, neutralizing antibodies were used at 10 µg/mL and refreshed daily until sample collection.

    Article Title: The ammonia oxidizing bacterium Nitrosomonas eutropha D23 blocks T helper 2 cell polarization via IL-10 – mediated interference with dendritic cell activation
    Article Snippet: AOB were added at a ratio of four bacteria per PBMC one hour prior to Th2 stimulation.AOB were added at a ratio of four bacteria per PBMC one hour prior to Th2 stimulation.. PBMC were then stimulated using either the CellXvivo human Th2 differentiation kit (R&D systems) following the manufacturer’s recommendations, or 1μg/mL SEB (Sigma).. Where appropriate, neutralizing antibodies were used at 10μg/mL and refreshed daily until sample collection.Where appropriate, neutralizing antibodies were used at 10μg/mL and refreshed daily until sample collection.

    Flow Cytometry:

    Article Title: The ammonia oxidizing bacterium Nitrosomonas eutropha blocks T helper 2 cell polarization via the anti-inflammatory cytokine IL-10
    Article Snippet: AOB were added at a ratio of four bacteria per PBMC 1 h prior to Th2 stimulation.AOB were added at a ratio of four bacteria per PBMC 1 h prior to Th2 stimulation.. PBMC were then stimulated using either the CellXvivo human Th2 differentiation kit (R&D systems) following the manufacturer’s recommendations, or 1 µg/mL SEB (Sigma).. Where appropriate, neutralizing antibodies were used at 10 µg/mL and refreshed daily until sample collection.Where appropriate, neutralizing antibodies were used at 10 µg/mL and refreshed daily until sample collection.

    Article Title: The ammonia oxidizing bacterium Nitrosomonas eutropha D23 blocks T helper 2 cell polarization via IL-10 – mediated interference with dendritic cell activation
    Article Snippet: AOB were added at a ratio of four bacteria per PBMC one hour prior to Th2 stimulation.AOB were added at a ratio of four bacteria per PBMC one hour prior to Th2 stimulation.. PBMC were then stimulated using either the CellXvivo human Th2 differentiation kit (R&D systems) following the manufacturer’s recommendations, or 1μg/mL SEB (Sigma).. Where appropriate, neutralizing antibodies were used at 10μg/mL and refreshed daily until sample collection.Where appropriate, neutralizing antibodies were used at 10μg/mL and refreshed daily until sample collection.

    Inhibition:

    Article Title: The ammonia oxidizing bacterium Nitrosomonas eutropha blocks T helper 2 cell polarization via the anti-inflammatory cytokine IL-10
    Article Snippet: AOB were added at a ratio of four bacteria per PBMC 1 h prior to Th2 stimulation.AOB were added at a ratio of four bacteria per PBMC 1 h prior to Th2 stimulation.. PBMC were then stimulated using either the CellXvivo human Th2 differentiation kit (R&D systems) following the manufacturer’s recommendations, or 1 µg/mL SEB (Sigma).. Where appropriate, neutralizing antibodies were used at 10 µg/mL and refreshed daily until sample collection.Where appropriate, neutralizing antibodies were used at 10 µg/mL and refreshed daily until sample collection.

    Article Title: The ammonia oxidizing bacterium Nitrosomonas eutropha D23 blocks T helper 2 cell polarization via IL-10 – mediated interference with dendritic cell activation
    Article Snippet: AOB were added at a ratio of four bacteria per PBMC one hour prior to Th2 stimulation.AOB were added at a ratio of four bacteria per PBMC one hour prior to Th2 stimulation.. PBMC were then stimulated using either the CellXvivo human Th2 differentiation kit (R&D systems) following the manufacturer’s recommendations, or 1μg/mL SEB (Sigma).. Where appropriate, neutralizing antibodies were used at 10μg/mL and refreshed daily until sample collection.Where appropriate, neutralizing antibodies were used at 10μg/mL and refreshed daily until sample collection.

    Staining:

    Article Title: The ammonia oxidizing bacterium Nitrosomonas eutropha blocks T helper 2 cell polarization via the anti-inflammatory cytokine IL-10
    Article Snippet: AOB were added at a ratio of four bacteria per PBMC 1 h prior to Th2 stimulation.AOB were added at a ratio of four bacteria per PBMC 1 h prior to Th2 stimulation.. PBMC were then stimulated using either the CellXvivo human Th2 differentiation kit (R&D systems) following the manufacturer’s recommendations, or 1 µg/mL SEB (Sigma).. Where appropriate, neutralizing antibodies were used at 10 µg/mL and refreshed daily until sample collection.Where appropriate, neutralizing antibodies were used at 10 µg/mL and refreshed daily until sample collection.

    Article Title: The ammonia oxidizing bacterium Nitrosomonas eutropha D23 blocks T helper 2 cell polarization via IL-10 – mediated interference with dendritic cell activation
    Article Snippet: AOB were added at a ratio of four bacteria per PBMC one hour prior to Th2 stimulation.AOB were added at a ratio of four bacteria per PBMC one hour prior to Th2 stimulation.. PBMC were then stimulated using either the CellXvivo human Th2 differentiation kit (R&D systems) following the manufacturer’s recommendations, or 1μg/mL SEB (Sigma).. Where appropriate, neutralizing antibodies were used at 10μg/mL and refreshed daily until sample collection.Where appropriate, neutralizing antibodies were used at 10μg/mL and refreshed daily until sample collection.

    Activity Assay:

    Article Title: The ammonia oxidizing bacterium Nitrosomonas eutropha blocks T helper 2 cell polarization via the anti-inflammatory cytokine IL-10
    Article Snippet: AOB were added at a ratio of four bacteria per PBMC 1 h prior to Th2 stimulation.AOB were added at a ratio of four bacteria per PBMC 1 h prior to Th2 stimulation.. PBMC were then stimulated using either the CellXvivo human Th2 differentiation kit (R&D systems) following the manufacturer’s recommendations, or 1 µg/mL SEB (Sigma).. Where appropriate, neutralizing antibodies were used at 10 µg/mL and refreshed daily until sample collection.Where appropriate, neutralizing antibodies were used at 10 µg/mL and refreshed daily until sample collection.

    Article Title: The ammonia oxidizing bacterium Nitrosomonas eutropha D23 blocks T helper 2 cell polarization via IL-10 – mediated interference with dendritic cell activation
    Article Snippet: AOB were added at a ratio of four bacteria per PBMC one hour prior to Th2 stimulation.AOB were added at a ratio of four bacteria per PBMC one hour prior to Th2 stimulation.. PBMC were then stimulated using either the CellXvivo human Th2 differentiation kit (R&D systems) following the manufacturer’s recommendations, or 1μg/mL SEB (Sigma).. Where appropriate, neutralizing antibodies were used at 10μg/mL and refreshed daily until sample collection.Where appropriate, neutralizing antibodies were used at 10μg/mL and refreshed daily until sample collection.

    Fluorescence:

    Article Title: The ammonia oxidizing bacterium Nitrosomonas eutropha blocks T helper 2 cell polarization via the anti-inflammatory cytokine IL-10
    Article Snippet: AOB were added at a ratio of four bacteria per PBMC 1 h prior to Th2 stimulation.AOB were added at a ratio of four bacteria per PBMC 1 h prior to Th2 stimulation.. PBMC were then stimulated using either the CellXvivo human Th2 differentiation kit (R&D systems) following the manufacturer’s recommendations, or 1 µg/mL SEB (Sigma).. Where appropriate, neutralizing antibodies were used at 10 µg/mL and refreshed daily until sample collection.Where appropriate, neutralizing antibodies were used at 10 µg/mL and refreshed daily until sample collection.

    Article Title: The ammonia oxidizing bacterium Nitrosomonas eutropha D23 blocks T helper 2 cell polarization via IL-10 – mediated interference with dendritic cell activation
    Article Snippet: AOB were added at a ratio of four bacteria per PBMC one hour prior to Th2 stimulation.AOB were added at a ratio of four bacteria per PBMC one hour prior to Th2 stimulation.. PBMC were then stimulated using either the CellXvivo human Th2 differentiation kit (R&D systems) following the manufacturer’s recommendations, or 1μg/mL SEB (Sigma).. Where appropriate, neutralizing antibodies were used at 10μg/mL and refreshed daily until sample collection.Where appropriate, neutralizing antibodies were used at 10μg/mL and refreshed daily until sample collection.

    Control:

    Article Title: The ammonia oxidizing bacterium Nitrosomonas eutropha blocks T helper 2 cell polarization via the anti-inflammatory cytokine IL-10
    Article Snippet: AOB were added at a ratio of four bacteria per PBMC 1 h prior to Th2 stimulation.AOB were added at a ratio of four bacteria per PBMC 1 h prior to Th2 stimulation.. PBMC were then stimulated using either the CellXvivo human Th2 differentiation kit (R&D systems) following the manufacturer’s recommendations, or 1 µg/mL SEB (Sigma).. Where appropriate, neutralizing antibodies were used at 10 µg/mL and refreshed daily until sample collection.Where appropriate, neutralizing antibodies were used at 10 µg/mL and refreshed daily until sample collection.

    Article Title: The ammonia oxidizing bacterium Nitrosomonas eutropha D23 blocks T helper 2 cell polarization via IL-10 – mediated interference with dendritic cell activation
    Article Snippet: AOB were added at a ratio of four bacteria per PBMC one hour prior to Th2 stimulation.AOB were added at a ratio of four bacteria per PBMC one hour prior to Th2 stimulation.. PBMC were then stimulated using either the CellXvivo human Th2 differentiation kit (R&D systems) following the manufacturer’s recommendations, or 1μg/mL SEB (Sigma).. Where appropriate, neutralizing antibodies were used at 10μg/mL and refreshed daily until sample collection.Where appropriate, neutralizing antibodies were used at 10μg/mL and refreshed daily until sample collection.

    Produced:

    Article Title: The ammonia oxidizing bacterium Nitrosomonas eutropha blocks T helper 2 cell polarization via the anti-inflammatory cytokine IL-10
    Article Snippet: AOB were added at a ratio of four bacteria per PBMC 1 h prior to Th2 stimulation.AOB were added at a ratio of four bacteria per PBMC 1 h prior to Th2 stimulation.. PBMC were then stimulated using either the CellXvivo human Th2 differentiation kit (R&D systems) following the manufacturer’s recommendations, or 1 µg/mL SEB (Sigma).. Where appropriate, neutralizing antibodies were used at 10 µg/mL and refreshed daily until sample collection.Where appropriate, neutralizing antibodies were used at 10 µg/mL and refreshed daily until sample collection.

    Article Title: The ammonia oxidizing bacterium Nitrosomonas eutropha D23 blocks T helper 2 cell polarization via IL-10 – mediated interference with dendritic cell activation
    Article Snippet: AOB were added at a ratio of four bacteria per PBMC one hour prior to Th2 stimulation.AOB were added at a ratio of four bacteria per PBMC one hour prior to Th2 stimulation.. PBMC were then stimulated using either the CellXvivo human Th2 differentiation kit (R&D systems) following the manufacturer’s recommendations, or 1μg/mL SEB (Sigma).. Where appropriate, neutralizing antibodies were used at 10μg/mL and refreshed daily until sample collection.Where appropriate, neutralizing antibodies were used at 10μg/mL and refreshed daily until sample collection.

    Incubation:

    Article Title: The ammonia oxidizing bacterium Nitrosomonas eutropha blocks T helper 2 cell polarization via the anti-inflammatory cytokine IL-10
    Article Snippet: AOB were added at a ratio of four bacteria per PBMC 1 h prior to Th2 stimulation.AOB were added at a ratio of four bacteria per PBMC 1 h prior to Th2 stimulation.. PBMC were then stimulated using either the CellXvivo human Th2 differentiation kit (R&D systems) following the manufacturer’s recommendations, or 1 µg/mL SEB (Sigma).. Where appropriate, neutralizing antibodies were used at 10 µg/mL and refreshed daily until sample collection.Where appropriate, neutralizing antibodies were used at 10 µg/mL and refreshed daily until sample collection.

    Article Title: The ammonia oxidizing bacterium Nitrosomonas eutropha D23 blocks T helper 2 cell polarization via IL-10 – mediated interference with dendritic cell activation
    Article Snippet: AOB were added at a ratio of four bacteria per PBMC one hour prior to Th2 stimulation.AOB were added at a ratio of four bacteria per PBMC one hour prior to Th2 stimulation.. PBMC were then stimulated using either the CellXvivo human Th2 differentiation kit (R&D systems) following the manufacturer’s recommendations, or 1μg/mL SEB (Sigma).. Where appropriate, neutralizing antibodies were used at 10μg/mL and refreshed daily until sample collection.Where appropriate, neutralizing antibodies were used at 10μg/mL and refreshed daily until sample collection.

    Cell Culture:

    Article Title: The ammonia oxidizing bacterium Nitrosomonas eutropha blocks T helper 2 cell polarization via the anti-inflammatory cytokine IL-10
    Article Snippet: AOB were added at a ratio of four bacteria per PBMC 1 h prior to Th2 stimulation.AOB were added at a ratio of four bacteria per PBMC 1 h prior to Th2 stimulation.. PBMC were then stimulated using either the CellXvivo human Th2 differentiation kit (R&D systems) following the manufacturer’s recommendations, or 1 µg/mL SEB (Sigma).. Where appropriate, neutralizing antibodies were used at 10 µg/mL and refreshed daily until sample collection.Where appropriate, neutralizing antibodies were used at 10 µg/mL and refreshed daily until sample collection.

    Article Title: The ammonia oxidizing bacterium Nitrosomonas eutropha D23 blocks T helper 2 cell polarization via IL-10 – mediated interference with dendritic cell activation
    Article Snippet: AOB were added at a ratio of four bacteria per PBMC one hour prior to Th2 stimulation.AOB were added at a ratio of four bacteria per PBMC one hour prior to Th2 stimulation.. PBMC were then stimulated using either the CellXvivo human Th2 differentiation kit (R&D systems) following the manufacturer’s recommendations, or 1μg/mL SEB (Sigma).. Where appropriate, neutralizing antibodies were used at 10μg/mL and refreshed daily until sample collection.Where appropriate, neutralizing antibodies were used at 10μg/mL and refreshed daily until sample collection.



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    AOB inhibit <t>Th2</t> immune polarization in human PBMC. ( a – f ) IL-5 and IL-13 supernatant levels are reduced with AOB pretreatment of PBMC prior to Th2 differentiation cocktail stimulation; measured by ELISA of supernatants 72 h post-stimulation. IL-4 gene expression is also reduced with AOB pretreatment; measured by qPCR 72 h post-stimulation. AOB pre-treatment prior to Th2 differentiation cocktail stimulation of PBMCs from donor A reduced the levels of IL-5 ( a ) and IL-13 ( b ) as well as IL-4 ( c ) gene expression (n ≥ 12, one-way ANOVA with multiple comparisons or unpaired t-test). ( d – f ) IL-5 ( d ), IL-13 ( e ), and IL-4 ( f ) levels/expression are reduced in AOB-pretreated PBMC from 3 to 5 different donors (n ≥ 6 replicates per donor, shown on the figure is unpaired t-test p value for individual donors, unpaired t-test p value for aggregated donors: p < 0.0001 for IL-5, p < 0.01 for IL-13, p < 0.001 for IL-4). ( g , h ) The number of CD4+ IL-5+ cells is reduced with AOB pretreatment of PBMC following Th2 differentiation cocktail stimulation; measured by flow cytometry with anti-CD4 PerCP cy5.5 and anti-IL-5 PE antibodies applied 72 h post-stimulation. ( g ) Representative dot plot from donor C cells showing the reduction in CD4+ and IL-5+ cells. ( h ) The percentage of Th2 cells as CD4+ IL-5+ are reduced with AOB pretreatment (n = 3, donor C, unpaired t-test). IL-5 ( i ) and IL-13 ( j ) supernatant levels are reduced in the presence of AOB 7 days post-stimulation with SEB (1 µg/mL), measured by ELISA (n = 3 per donor, 3 donors, unpaired t-test, p < 0.0001 for IL-5 and IL-13 for aggregated donor data sets).
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    Image Search Results


    Fig. 1. The steroid resistance in the OVA/IL-33–induced asthma model was associated with the acquisition of steroid resistance by ILC2s but not TH2 cells. (A) Schematic diagram of the asthma models induced by OVA alone and OVA/IL-33. (B and C) AHR of the OVA-alone (B) and OVA/IL-33 (C) models with or without Dex treatment. (D and E) Immune-cell numbers in the BALF (D) and H&E staining of the lungs (E) of the OVA-alone and OVA/IL-33 models with or without Dex treatment. Scale bars, 100 μm. (F to J) Frequencies of IL-5– and IL-13–producing CD4+ T cells (gated as CD45+Lineage+CD4+ cells) and ILCs (gated as CD45+Lineage−CD90.2+ cells) in the OVA-alone and OVA/IL-33 models with or without Dex treatment. Representative dot plots are shown in (F) and the frequencies of IL-5+ (G) and IL-13+ (H) CD4+ T cells and IL-5+ (I) and IL-13+ (J) ILC2s are plotted. *P ≤0.05, **P ≤0.01, ***P ≤0.001, and ****P ≤0.0001, by one-way ANOVA followed by Bonferroni’s posttest. The data are representative of two to three independent experiments and are presented as mean ± SEM.

    Journal: Science advances

    Article Title: JAK3 inhibitor suppresses multipotent ILC2s and attenuates steroid-resistant asthma.

    doi: 10.1126/sciadv.adi3770

    Figure Lengend Snippet: Fig. 1. The steroid resistance in the OVA/IL-33–induced asthma model was associated with the acquisition of steroid resistance by ILC2s but not TH2 cells. (A) Schematic diagram of the asthma models induced by OVA alone and OVA/IL-33. (B and C) AHR of the OVA-alone (B) and OVA/IL-33 (C) models with or without Dex treatment. (D and E) Immune-cell numbers in the BALF (D) and H&E staining of the lungs (E) of the OVA-alone and OVA/IL-33 models with or without Dex treatment. Scale bars, 100 μm. (F to J) Frequencies of IL-5– and IL-13–producing CD4+ T cells (gated as CD45+Lineage+CD4+ cells) and ILCs (gated as CD45+Lineage−CD90.2+ cells) in the OVA-alone and OVA/IL-33 models with or without Dex treatment. Representative dot plots are shown in (F) and the frequencies of IL-5+ (G) and IL-13+ (H) CD4+ T cells and IL-5+ (I) and IL-13+ (J) ILC2s are plotted. *P ≤0.05, **P ≤0.01, ***P ≤0.001, and ****P ≤0.0001, by one-way ANOVA followed by Bonferroni’s posttest. The data are representative of two to three independent experiments and are presented as mean ± SEM.

    Article Snippet: Human naïve CD4+ T cells were isolated with the Mojosort Human CD4 Naive T cell Isolation Kit (BioLegend, CA, USA) and differentiated to TH2 cells using CellXVivo Human TH2 Cell Differentiation Kit (R&D systems, MN, USA).

    Techniques: Staining

    Fig. 3. IL-2/7/33 treatment induces steroid-resistant murine and human ILC2s in vitro but TH2 cells remain steroid sensitive. (A) Schematic diagram showing the isolation of murine lung ILC2s and differentiation of TH2 cells from naïve murine splenocytes in vitro, and their treatment with IL-2/7/33 with and without Dex. (B and C) Annexin V expression by the murine ILC2s (B) and TH2 cells (C). (D to G) Frequencies of IL-5– and IL-13–producing murine ILC2s [(D) and (E)] and TH2 cells [(F) and (G)]. Representative dot plots are shown in (D) and (F). The frequencies of IL-5+ and IL-13+ cells are plotted in (E) and (G). (H) Schematic diagram of the isolation of ILCs and CD4+ T cells from human PBMCs (n = 10 subjects) and their in vitro treatment with IL-2/7/33 with and without Dex. (I to J) Frequencies of IL-5+ and IL-13+ ILCs (I) and TH2 cells (J) are plotted. n.s., not significant; *P ≤0.05, **P ≤0.01, and ****P ≤0.0001, by unpaired t test [(B) and (C)], one-way ANOVA followed by Bonferroni’s posttest [(E) and (G)], and paired t test [(I) and (J)]. The data are representative of two to three independent experiments and are presented as mean ± SEM.

    Journal: Science advances

    Article Title: JAK3 inhibitor suppresses multipotent ILC2s and attenuates steroid-resistant asthma.

    doi: 10.1126/sciadv.adi3770

    Figure Lengend Snippet: Fig. 3. IL-2/7/33 treatment induces steroid-resistant murine and human ILC2s in vitro but TH2 cells remain steroid sensitive. (A) Schematic diagram showing the isolation of murine lung ILC2s and differentiation of TH2 cells from naïve murine splenocytes in vitro, and their treatment with IL-2/7/33 with and without Dex. (B and C) Annexin V expression by the murine ILC2s (B) and TH2 cells (C). (D to G) Frequencies of IL-5– and IL-13–producing murine ILC2s [(D) and (E)] and TH2 cells [(F) and (G)]. Representative dot plots are shown in (D) and (F). The frequencies of IL-5+ and IL-13+ cells are plotted in (E) and (G). (H) Schematic diagram of the isolation of ILCs and CD4+ T cells from human PBMCs (n = 10 subjects) and their in vitro treatment with IL-2/7/33 with and without Dex. (I to J) Frequencies of IL-5+ and IL-13+ ILCs (I) and TH2 cells (J) are plotted. n.s., not significant; *P ≤0.05, **P ≤0.01, and ****P ≤0.0001, by unpaired t test [(B) and (C)], one-way ANOVA followed by Bonferroni’s posttest [(E) and (G)], and paired t test [(I) and (J)]. The data are representative of two to three independent experiments and are presented as mean ± SEM.

    Article Snippet: Human naïve CD4+ T cells were isolated with the Mojosort Human CD4 Naive T cell Isolation Kit (BioLegend, CA, USA) and differentiated to TH2 cells using CellXVivo Human TH2 Cell Differentiation Kit (R&D systems, MN, USA).

    Techniques: In Vitro, Isolation, Expressing

    AOB inhibit Th2 immune polarization in human PBMC. ( a – f ) IL-5 and IL-13 supernatant levels are reduced with AOB pretreatment of PBMC prior to Th2 differentiation cocktail stimulation; measured by ELISA of supernatants 72 h post-stimulation. IL-4 gene expression is also reduced with AOB pretreatment; measured by qPCR 72 h post-stimulation. AOB pre-treatment prior to Th2 differentiation cocktail stimulation of PBMCs from donor A reduced the levels of IL-5 ( a ) and IL-13 ( b ) as well as IL-4 ( c ) gene expression (n ≥ 12, one-way ANOVA with multiple comparisons or unpaired t-test). ( d – f ) IL-5 ( d ), IL-13 ( e ), and IL-4 ( f ) levels/expression are reduced in AOB-pretreated PBMC from 3 to 5 different donors (n ≥ 6 replicates per donor, shown on the figure is unpaired t-test p value for individual donors, unpaired t-test p value for aggregated donors: p < 0.0001 for IL-5, p < 0.01 for IL-13, p < 0.001 for IL-4). ( g , h ) The number of CD4+ IL-5+ cells is reduced with AOB pretreatment of PBMC following Th2 differentiation cocktail stimulation; measured by flow cytometry with anti-CD4 PerCP cy5.5 and anti-IL-5 PE antibodies applied 72 h post-stimulation. ( g ) Representative dot plot from donor C cells showing the reduction in CD4+ and IL-5+ cells. ( h ) The percentage of Th2 cells as CD4+ IL-5+ are reduced with AOB pretreatment (n = 3, donor C, unpaired t-test). IL-5 ( i ) and IL-13 ( j ) supernatant levels are reduced in the presence of AOB 7 days post-stimulation with SEB (1 µg/mL), measured by ELISA (n = 3 per donor, 3 donors, unpaired t-test, p < 0.0001 for IL-5 and IL-13 for aggregated donor data sets).

    Journal: Scientific Reports

    Article Title: The ammonia oxidizing bacterium Nitrosomonas eutropha blocks T helper 2 cell polarization via the anti-inflammatory cytokine IL-10

    doi: 10.1038/s41598-021-93299-1

    Figure Lengend Snippet: AOB inhibit Th2 immune polarization in human PBMC. ( a – f ) IL-5 and IL-13 supernatant levels are reduced with AOB pretreatment of PBMC prior to Th2 differentiation cocktail stimulation; measured by ELISA of supernatants 72 h post-stimulation. IL-4 gene expression is also reduced with AOB pretreatment; measured by qPCR 72 h post-stimulation. AOB pre-treatment prior to Th2 differentiation cocktail stimulation of PBMCs from donor A reduced the levels of IL-5 ( a ) and IL-13 ( b ) as well as IL-4 ( c ) gene expression (n ≥ 12, one-way ANOVA with multiple comparisons or unpaired t-test). ( d – f ) IL-5 ( d ), IL-13 ( e ), and IL-4 ( f ) levels/expression are reduced in AOB-pretreated PBMC from 3 to 5 different donors (n ≥ 6 replicates per donor, shown on the figure is unpaired t-test p value for individual donors, unpaired t-test p value for aggregated donors: p < 0.0001 for IL-5, p < 0.01 for IL-13, p < 0.001 for IL-4). ( g , h ) The number of CD4+ IL-5+ cells is reduced with AOB pretreatment of PBMC following Th2 differentiation cocktail stimulation; measured by flow cytometry with anti-CD4 PerCP cy5.5 and anti-IL-5 PE antibodies applied 72 h post-stimulation. ( g ) Representative dot plot from donor C cells showing the reduction in CD4+ and IL-5+ cells. ( h ) The percentage of Th2 cells as CD4+ IL-5+ are reduced with AOB pretreatment (n = 3, donor C, unpaired t-test). IL-5 ( i ) and IL-13 ( j ) supernatant levels are reduced in the presence of AOB 7 days post-stimulation with SEB (1 µg/mL), measured by ELISA (n = 3 per donor, 3 donors, unpaired t-test, p < 0.0001 for IL-5 and IL-13 for aggregated donor data sets).

    Article Snippet: PBMC were then stimulated using either the CellXvivo human Th2 differentiation kit (R&D systems) following the manufacturer’s recommendations, or 1 μg/mL SEB (Sigma).

    Techniques: Enzyme-linked Immunosorbent Assay, Gene Expression, Expressing, Flow Cytometry

    AOB induce Th1 polarization but Th1 are not necessary for Th2 inhibition. ( a – d ) IFNγ ( a , b ) and IL-12p70 ( c , d ) supernatant levels are increased with AOB pretreatment of PBMC prior to Th2 kit stimulation; measured by ELISA. IFNγ ( a ) and IL-12p70 ( c ) levels are increased in PBMC from donor A (n = 9, one-way ANOVA with multiple comparisons). IFNγ ( b ) is induced in AOB pre-treated PBMC from 5 donors and IL-12p70 ( d ) is induced in PBMC from 2 of 3 donors tested (n ≥ 6 per donor, unpaired t-test, p < 0.0001 for IFNγ and p < 0.0015 for IL-12 aggregated donor datasets). ( e , f ) The number of CD4+ IFNγ+ cells is increased with AOB pretreatment of PBMC prior to Th2 differentiation cocktail stimulation as measured by staining with anti-CD4 PerCP cy5.5 and anti-IFNγ FITC antibodies 72 h post-stimulation. ( e ) Representative dot plot from donor C PBMC showing an increase in the percentage of CD4+ and IFNγ+ cells. ( f ) Percentage of Th1 cells as CD4+ IFNγ+ are increased with AOB pretreatment (n = 6, unpaired t-test). ( g , h ) Addition of IFNγ or IL-12 neutralizing antibodies to PBMC prior to AOB pretreatment does not prevent AOB inhibition of Th2. IFNγ ( g ) production is reduced in presence of 10 µg/mL IFNγ or IL-12 neutralizing antibodies in PBMC from 3 donors (n = 3 per donor, one point per donor, one-way ANOVA with multiple comparisons). AOB-mediated reduction in IL-5 production ( h ) is unaffected in presence of 10 µg/mL IFNγ or IL-12 neutralizing antibodies in PBMC from 3 donors (n = 3 per donor, one point per donor, one-way ANOVA with multiple comparisons).

    Journal: Scientific Reports

    Article Title: The ammonia oxidizing bacterium Nitrosomonas eutropha blocks T helper 2 cell polarization via the anti-inflammatory cytokine IL-10

    doi: 10.1038/s41598-021-93299-1

    Figure Lengend Snippet: AOB induce Th1 polarization but Th1 are not necessary for Th2 inhibition. ( a – d ) IFNγ ( a , b ) and IL-12p70 ( c , d ) supernatant levels are increased with AOB pretreatment of PBMC prior to Th2 kit stimulation; measured by ELISA. IFNγ ( a ) and IL-12p70 ( c ) levels are increased in PBMC from donor A (n = 9, one-way ANOVA with multiple comparisons). IFNγ ( b ) is induced in AOB pre-treated PBMC from 5 donors and IL-12p70 ( d ) is induced in PBMC from 2 of 3 donors tested (n ≥ 6 per donor, unpaired t-test, p < 0.0001 for IFNγ and p < 0.0015 for IL-12 aggregated donor datasets). ( e , f ) The number of CD4+ IFNγ+ cells is increased with AOB pretreatment of PBMC prior to Th2 differentiation cocktail stimulation as measured by staining with anti-CD4 PerCP cy5.5 and anti-IFNγ FITC antibodies 72 h post-stimulation. ( e ) Representative dot plot from donor C PBMC showing an increase in the percentage of CD4+ and IFNγ+ cells. ( f ) Percentage of Th1 cells as CD4+ IFNγ+ are increased with AOB pretreatment (n = 6, unpaired t-test). ( g , h ) Addition of IFNγ or IL-12 neutralizing antibodies to PBMC prior to AOB pretreatment does not prevent AOB inhibition of Th2. IFNγ ( g ) production is reduced in presence of 10 µg/mL IFNγ or IL-12 neutralizing antibodies in PBMC from 3 donors (n = 3 per donor, one point per donor, one-way ANOVA with multiple comparisons). AOB-mediated reduction in IL-5 production ( h ) is unaffected in presence of 10 µg/mL IFNγ or IL-12 neutralizing antibodies in PBMC from 3 donors (n = 3 per donor, one point per donor, one-way ANOVA with multiple comparisons).

    Article Snippet: PBMC were then stimulated using either the CellXvivo human Th2 differentiation kit (R&D systems) following the manufacturer’s recommendations, or 1 μg/mL SEB (Sigma).

    Techniques: Inhibition, Enzyme-linked Immunosorbent Assay, Staining

    AOB-mediated Th2 inhibition requires IL-10 and is associated with a suppression of dendritic cell activity. ( a – d ) Expression of MHC II ( a , b ) or CD86 ( c , d ) in CD11c+ cells is reduced in PBMC treated with AOB prior to Th2 differentiation cocktail stimulation; measured by flow cytometry in Th2-stimulated PBMC in the presence or absence of AOB. ( a , c ) Representative fluorescence intensity plots of anti-MHC II FITC ( a ) or anti-CD86 PE ( c ) stained PBMC from donor C in the presence (red) or absence (grey) of AOB. ( b , d ) Relative MHC II ( b ) or CD86 ( d ) expression in CD11c+ positive cells is reduced with AOB treatment in 3 donors (n = 3 per donor, unpaired t-test, p < 0.0001 for MHC II and CD86 aggregated donor datasets). ( e ) IL-10 production is induced by AOB in PBMC from 3 donors 24 h post-stimulation; measured by ELISA of culture supernatants (n = 3 per donor, p = 0.1095 for aggregated donor data set). ( f – h ) Addition of IL-10 neutralizing antibodies to PBMC prior to AOB pretreatment interfered with AOB’s inhibition of Th2. IL-10 ( f ) production by Th2-stimulated PBMC was reduced with the addition of IL-10 neutralizing antibody but not with isotype control in 3 donors (n = 3 per donor, one point per donor, one-way ANOVA with multiple comparisons). AOB-mediated fold reduction in IL-5 ( g ) was obstructed by the addition of IL-10 neutralizing antibody compared to isotype control in 2 of 3 donors tested (n = 3 per donor, unpaired t test). AOB-mediated reduction in CD86 expression ( h ) in CD11c+ cells is hindered with the addition of IL-10 neutralizing antibody but not with isotype control (n = 3, donor D, one-way ANOVA with multiple comparisons).

    Journal: Scientific Reports

    Article Title: The ammonia oxidizing bacterium Nitrosomonas eutropha blocks T helper 2 cell polarization via the anti-inflammatory cytokine IL-10

    doi: 10.1038/s41598-021-93299-1

    Figure Lengend Snippet: AOB-mediated Th2 inhibition requires IL-10 and is associated with a suppression of dendritic cell activity. ( a – d ) Expression of MHC II ( a , b ) or CD86 ( c , d ) in CD11c+ cells is reduced in PBMC treated with AOB prior to Th2 differentiation cocktail stimulation; measured by flow cytometry in Th2-stimulated PBMC in the presence or absence of AOB. ( a , c ) Representative fluorescence intensity plots of anti-MHC II FITC ( a ) or anti-CD86 PE ( c ) stained PBMC from donor C in the presence (red) or absence (grey) of AOB. ( b , d ) Relative MHC II ( b ) or CD86 ( d ) expression in CD11c+ positive cells is reduced with AOB treatment in 3 donors (n = 3 per donor, unpaired t-test, p < 0.0001 for MHC II and CD86 aggregated donor datasets). ( e ) IL-10 production is induced by AOB in PBMC from 3 donors 24 h post-stimulation; measured by ELISA of culture supernatants (n = 3 per donor, p = 0.1095 for aggregated donor data set). ( f – h ) Addition of IL-10 neutralizing antibodies to PBMC prior to AOB pretreatment interfered with AOB’s inhibition of Th2. IL-10 ( f ) production by Th2-stimulated PBMC was reduced with the addition of IL-10 neutralizing antibody but not with isotype control in 3 donors (n = 3 per donor, one point per donor, one-way ANOVA with multiple comparisons). AOB-mediated fold reduction in IL-5 ( g ) was obstructed by the addition of IL-10 neutralizing antibody compared to isotype control in 2 of 3 donors tested (n = 3 per donor, unpaired t test). AOB-mediated reduction in CD86 expression ( h ) in CD11c+ cells is hindered with the addition of IL-10 neutralizing antibody but not with isotype control (n = 3, donor D, one-way ANOVA with multiple comparisons).

    Article Snippet: PBMC were then stimulated using either the CellXvivo human Th2 differentiation kit (R&D systems) following the manufacturer’s recommendations, or 1 μg/mL SEB (Sigma).

    Techniques: Inhibition, Activity Assay, Expressing, Flow Cytometry, Fluorescence, Staining, Enzyme-linked Immunosorbent Assay, Control

    Structural components could be responsible for AOB-mediated Th2 inhibition. ( a , b ) AOB metabolites nitrite ( a ) and nitric oxide ( b ) are produced by live but not heat killed AOB after 1 h incubation in AOB culture media (n = 3, unpaired t-test). ( c , d ) Fold-reduction in IL-5 ( c ) and IL-13 ( d ) mediated by AOB (red) is not significantly different from fold-reduction mediated by heat killed AOB (blue); measured by ELISA in cell culture supernatant collected 72 h post-stimulation with the Th2 stimulation cocktail (n ≥ 6 per donor, 5 donors, one datapoint per donor, unpaired T test). ( e , f ) Fold reduction in MHC II ( e ) and CD86 ( f ) expression in CD11c+ cells mediated by live AOB (red) is not significantly different from fold-reduction mediated by heat killed AOB (blue); measured by flow cytometry 72 h after Th2 stimulation in donor C (n = 6, unpaired T test). ( g , h ) IFNγ ( g ) and IL-12p70 ( h ) production by PBMC are not as strongly induced by heat killed AOB (blue) as by live AOB (red); measured by ELISA from supernatant of PBMC culture collected 72 h post-stimulation (n ≥ 3 per donor, 3 or 5 donors, one datapoint per donor, one-way ANOVA with multiple comparisons).

    Journal: Scientific Reports

    Article Title: The ammonia oxidizing bacterium Nitrosomonas eutropha blocks T helper 2 cell polarization via the anti-inflammatory cytokine IL-10

    doi: 10.1038/s41598-021-93299-1

    Figure Lengend Snippet: Structural components could be responsible for AOB-mediated Th2 inhibition. ( a , b ) AOB metabolites nitrite ( a ) and nitric oxide ( b ) are produced by live but not heat killed AOB after 1 h incubation in AOB culture media (n = 3, unpaired t-test). ( c , d ) Fold-reduction in IL-5 ( c ) and IL-13 ( d ) mediated by AOB (red) is not significantly different from fold-reduction mediated by heat killed AOB (blue); measured by ELISA in cell culture supernatant collected 72 h post-stimulation with the Th2 stimulation cocktail (n ≥ 6 per donor, 5 donors, one datapoint per donor, unpaired T test). ( e , f ) Fold reduction in MHC II ( e ) and CD86 ( f ) expression in CD11c+ cells mediated by live AOB (red) is not significantly different from fold-reduction mediated by heat killed AOB (blue); measured by flow cytometry 72 h after Th2 stimulation in donor C (n = 6, unpaired T test). ( g , h ) IFNγ ( g ) and IL-12p70 ( h ) production by PBMC are not as strongly induced by heat killed AOB (blue) as by live AOB (red); measured by ELISA from supernatant of PBMC culture collected 72 h post-stimulation (n ≥ 3 per donor, 3 or 5 donors, one datapoint per donor, one-way ANOVA with multiple comparisons).

    Article Snippet: PBMC were then stimulated using either the CellXvivo human Th2 differentiation kit (R&D systems) following the manufacturer’s recommendations, or 1 μg/mL SEB (Sigma).

    Techniques: Inhibition, Produced, Incubation, Enzyme-linked Immunosorbent Assay, Cell Culture, Expressing, Flow Cytometry