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Impact of gonococcal OMVs on dendritic cell activation and T cell differentiation. Mature murine bone marrow-derived dendritic cells (BMDCs) were stimulated with OMV-PorB WT , OMV-PorB K117Q/K171Q or a Mock control and surface expression of CD86 and <t>MHCII</t> was determined by flow cytometry. Stimulated BMDCs were subsequently co-cultured with naïve murine T cells and IFN-γ, TNF-α, and IL-4 cytokine levels in the culture supernatant was determined by ELISA. (A) Representative flow cytometry repeat for detection of CD86 levels at the surface of OMV-stimulated DCs. (B) Quantification of CD86 levels at the surface of OMV-stimulated DCs. (C) Representative flow cytometry repeat for detection of MHCII levels at the surface of OMV-stimulated DCs. (D) Quantification of MHCII levels at the surface of OMV-stimulated DCs. (E) Quantification of IL-2 secretion in the culture supernatant after co-culture of T cells with OMV-stimulated DCs. (F) Quantification of IL-10 secretion in the culture supernatant after co-culture of T cells with OMV-stimulated DCs. (G) Quantification of IL-4 secretion in the culture supernatant after co-culture of T cells with OMV-stimulated DCs. (H) Quantification of IFN-γ secretion in the culture supernatant after co-culture of T cells with OMV-stimulated DCs. (I) Quantification of TNF-α secretion in the culture supernatant after co-culture of T cells with OMV-stimulated DCs. (J) Quantification of IL-17A secretion in the culture supernatant after co-culture of T cells with OMV-stimulated DCs. Graphs represent mean ± SD of three biological repeats. Significant differences were identified by one-way ANOVA (GraphPad Prism).
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Impact of gonococcal OMVs on dendritic cell activation and T cell differentiation. Mature murine bone marrow-derived dendritic cells (BMDCs) were stimulated with OMV-PorB WT , OMV-PorB K117Q/K171Q or a Mock control and surface expression of CD86 and <t>MHCII</t> was determined by flow cytometry. Stimulated BMDCs were subsequently co-cultured with naïve murine T cells and IFN-γ, TNF-α, and IL-4 cytokine levels in the culture supernatant was determined by ELISA. (A) Representative flow cytometry repeat for detection of CD86 levels at the surface of OMV-stimulated DCs. (B) Quantification of CD86 levels at the surface of OMV-stimulated DCs. (C) Representative flow cytometry repeat for detection of MHCII levels at the surface of OMV-stimulated DCs. (D) Quantification of MHCII levels at the surface of OMV-stimulated DCs. (E) Quantification of IL-2 secretion in the culture supernatant after co-culture of T cells with OMV-stimulated DCs. (F) Quantification of IL-10 secretion in the culture supernatant after co-culture of T cells with OMV-stimulated DCs. (G) Quantification of IL-4 secretion in the culture supernatant after co-culture of T cells with OMV-stimulated DCs. (H) Quantification of IFN-γ secretion in the culture supernatant after co-culture of T cells with OMV-stimulated DCs. (I) Quantification of TNF-α secretion in the culture supernatant after co-culture of T cells with OMV-stimulated DCs. (J) Quantification of IL-17A secretion in the culture supernatant after co-culture of T cells with OMV-stimulated DCs. Graphs represent mean ± SD of three biological repeats. Significant differences were identified by one-way ANOVA (GraphPad Prism).
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Impact of gonococcal OMVs on dendritic cell activation and T cell differentiation. Mature murine bone marrow-derived dendritic cells (BMDCs) were stimulated with OMV-PorB WT , OMV-PorB K117Q/K171Q or a Mock control and surface expression of CD86 and MHCII was determined by flow cytometry. Stimulated BMDCs were subsequently co-cultured with naïve murine T cells and IFN-γ, TNF-α, and IL-4 cytokine levels in the culture supernatant was determined by ELISA. (A) Representative flow cytometry repeat for detection of CD86 levels at the surface of OMV-stimulated DCs. (B) Quantification of CD86 levels at the surface of OMV-stimulated DCs. (C) Representative flow cytometry repeat for detection of MHCII levels at the surface of OMV-stimulated DCs. (D) Quantification of MHCII levels at the surface of OMV-stimulated DCs. (E) Quantification of IL-2 secretion in the culture supernatant after co-culture of T cells with OMV-stimulated DCs. (F) Quantification of IL-10 secretion in the culture supernatant after co-culture of T cells with OMV-stimulated DCs. (G) Quantification of IL-4 secretion in the culture supernatant after co-culture of T cells with OMV-stimulated DCs. (H) Quantification of IFN-γ secretion in the culture supernatant after co-culture of T cells with OMV-stimulated DCs. (I) Quantification of TNF-α secretion in the culture supernatant after co-culture of T cells with OMV-stimulated DCs. (J) Quantification of IL-17A secretion in the culture supernatant after co-culture of T cells with OMV-stimulated DCs. Graphs represent mean ± SD of three biological repeats. Significant differences were identified by one-way ANOVA (GraphPad Prism).

Journal: Emerging Microbes & Infections

Article Title: Abolishing PorB-induced mitophagy enhances gonococcal outer membrane vesicle vaccine efficacy

doi: 10.1080/22221751.2026.2651468

Figure Lengend Snippet: Impact of gonococcal OMVs on dendritic cell activation and T cell differentiation. Mature murine bone marrow-derived dendritic cells (BMDCs) were stimulated with OMV-PorB WT , OMV-PorB K117Q/K171Q or a Mock control and surface expression of CD86 and MHCII was determined by flow cytometry. Stimulated BMDCs were subsequently co-cultured with naïve murine T cells and IFN-γ, TNF-α, and IL-4 cytokine levels in the culture supernatant was determined by ELISA. (A) Representative flow cytometry repeat for detection of CD86 levels at the surface of OMV-stimulated DCs. (B) Quantification of CD86 levels at the surface of OMV-stimulated DCs. (C) Representative flow cytometry repeat for detection of MHCII levels at the surface of OMV-stimulated DCs. (D) Quantification of MHCII levels at the surface of OMV-stimulated DCs. (E) Quantification of IL-2 secretion in the culture supernatant after co-culture of T cells with OMV-stimulated DCs. (F) Quantification of IL-10 secretion in the culture supernatant after co-culture of T cells with OMV-stimulated DCs. (G) Quantification of IL-4 secretion in the culture supernatant after co-culture of T cells with OMV-stimulated DCs. (H) Quantification of IFN-γ secretion in the culture supernatant after co-culture of T cells with OMV-stimulated DCs. (I) Quantification of TNF-α secretion in the culture supernatant after co-culture of T cells with OMV-stimulated DCs. (J) Quantification of IL-17A secretion in the culture supernatant after co-culture of T cells with OMV-stimulated DCs. Graphs represent mean ± SD of three biological repeats. Significant differences were identified by one-way ANOVA (GraphPad Prism).

Article Snippet: Mature DCs (1 × 106 cells/well) were stimulated for 20 h with 20 μg of OMV-PorB WT or OMV-PorB K117Q/K171Q or with a PBS-only Mock control. for 20 h. Expression of CD86 and MHCII was analyzed by flow cytometry on CD11c-positive cells (FITC-labeled anti-mouse CD11c; Elabscience, #E-AB-F0991C) using PE-labeled anti-mouse CD86 (Elabscience, #E-AB-F0994D) and APC-labeled anti-mouse MHCII (Elabscience, #E-AB-F0990E).

Techniques: Activation Assay, Cell Differentiation, Derivative Assay, Control, Expressing, Flow Cytometry, Cell Culture, Enzyme-linked Immunosorbent Assay, Co-Culture Assay