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leaf purified anti-tgf-β neutralizing antibody (clone 2d11.16.8)  (Bio X Cell)


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    Bio X Cell leaf purified anti-tgf-β neutralizing antibody (clone 2d11.16.8)
    Cell sorting strategy, phenotypic and functional characterization of CpG-induced c-kit + Sca-1 + B220 int PDCA-1 + BM cells. ( a ) Total BM cells incubated with PBS or CpG-B (1 µg/ml) for 18 h and magnetically enriched for c-kit + cells were further labelled for Sca-1, B220, PDCA-1 and electronically sorted into c-kit + Sca-1 + B220 int PDCA-1 + cells. Flow cytometry dot plots representative of 30 experiments. ( b ) Further characterization of cell-sorted CpG- as well as PBS-induced BM population was performed using flow cytometry analysis of the expression of various myeloid and dendritic cell markers as well as cell migration receptors. Specific antibody staining is depicted with open histograms (blue line for PBS- and red line for CpG-induced cells). Positive cells are defined using FMO controls (grey histograms). Two experiments. ( c ) FACS analysis of intranuclear expression of transcription factors. Analysis of IRF8 and Id2 expression with specific antibodies compared to FMO controls and of E2.2 and PU.1 by flowRNA, compared with B220 + PDCA-1 − control B cells (grey histograms), in CpG- versus PBS-stimulated c-kit + Sca-1 + B220 int PDCA-1 + population. Two experiments. ( d ) Frequency of c-kit + Sca-1 + B220 int PDCA-1 + cells emerging among total BM cells after 18 h of incubation with different TLR agonists, measured in BM cell cultures of four individual mice. Statistical significance as indicated, analyzed by Mann–Whitney test. ( e ) Cytokine production of cell-sorted PBS- versus CpG-induced c-kit + Sca-1 + B220 int PDCA-1 + cells, measured in the supernatant after 4 h of activation with PMA/ionomycin in the presence of brefeldin A using multiplex ELISA. Biological duplicates. ( f ) Intracytoplasmic <t>TGF-β</t> expression was analyzed by FACS in PBS- and CpG-pre-pDCs. One experiment out of two ( g ) IFN-α production was measured by ELISA in supernatants of CpG- versus PBS-induced cell-sorted c-kit + Sca-1 + B220 int PDCA-1 + BM cells after 18 h incubation with 1 μg/ml TLR-7 (R848), TLR-9 (CpG-B and CpG-P) and TLR-4 (LPS) agonists (biological duplicates). ( h ) Mature spleen pDCs and CpG-pre-pDCs were compared for their IFN-α response after 18-h incubation with CpG-B (1 μg/ml), measured by ELISA in supernatants (biological duplicates).
    Leaf Purified Anti Tgf β Neutralizing Antibody (Clone 2d11.16.8), supplied by Bio X Cell, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/leaf+purified+anti-tgf-%CE%B2+neutralizing+antibody+(clone+2d11%2E16%2E8)/leaf+purified+anti+tgf+%CE%B2+neutralizing+antibody++clone+2d11+16+8+/pmc07910458-219-15-22
    Average 90 stars, based on 1 article reviews
    leaf purified anti-tgf-β neutralizing antibody (clone 2d11.16.8) - by Bioz Stars, 2026-09
    90/100 stars

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    1) Product Images from "Toll-like receptor-9 stimulated plasmacytoid dendritic cell precursors suppress autoimmune neuroinflammation in a murine model of multiple sclerosis"

    Article Title: Toll-like receptor-9 stimulated plasmacytoid dendritic cell precursors suppress autoimmune neuroinflammation in a murine model of multiple sclerosis

    Journal: Scientific Reports

    doi: 10.1038/s41598-021-84023-0

    Cell sorting strategy, phenotypic and functional characterization of CpG-induced c-kit + Sca-1 + B220 int PDCA-1 + BM cells. ( a ) Total BM cells incubated with PBS or CpG-B (1 µg/ml) for 18 h and magnetically enriched for c-kit + cells were further labelled for Sca-1, B220, PDCA-1 and electronically sorted into c-kit + Sca-1 + B220 int PDCA-1 + cells. Flow cytometry dot plots representative of 30 experiments. ( b ) Further characterization of cell-sorted CpG- as well as PBS-induced BM population was performed using flow cytometry analysis of the expression of various myeloid and dendritic cell markers as well as cell migration receptors. Specific antibody staining is depicted with open histograms (blue line for PBS- and red line for CpG-induced cells). Positive cells are defined using FMO controls (grey histograms). Two experiments. ( c ) FACS analysis of intranuclear expression of transcription factors. Analysis of IRF8 and Id2 expression with specific antibodies compared to FMO controls and of E2.2 and PU.1 by flowRNA, compared with B220 + PDCA-1 − control B cells (grey histograms), in CpG- versus PBS-stimulated c-kit + Sca-1 + B220 int PDCA-1 + population. Two experiments. ( d ) Frequency of c-kit + Sca-1 + B220 int PDCA-1 + cells emerging among total BM cells after 18 h of incubation with different TLR agonists, measured in BM cell cultures of four individual mice. Statistical significance as indicated, analyzed by Mann–Whitney test. ( e ) Cytokine production of cell-sorted PBS- versus CpG-induced c-kit + Sca-1 + B220 int PDCA-1 + cells, measured in the supernatant after 4 h of activation with PMA/ionomycin in the presence of brefeldin A using multiplex ELISA. Biological duplicates. ( f ) Intracytoplasmic TGF-β expression was analyzed by FACS in PBS- and CpG-pre-pDCs. One experiment out of two ( g ) IFN-α production was measured by ELISA in supernatants of CpG- versus PBS-induced cell-sorted c-kit + Sca-1 + B220 int PDCA-1 + BM cells after 18 h incubation with 1 μg/ml TLR-7 (R848), TLR-9 (CpG-B and CpG-P) and TLR-4 (LPS) agonists (biological duplicates). ( h ) Mature spleen pDCs and CpG-pre-pDCs were compared for their IFN-α response after 18-h incubation with CpG-B (1 μg/ml), measured by ELISA in supernatants (biological duplicates).
    Figure Legend Snippet: Cell sorting strategy, phenotypic and functional characterization of CpG-induced c-kit + Sca-1 + B220 int PDCA-1 + BM cells. ( a ) Total BM cells incubated with PBS or CpG-B (1 µg/ml) for 18 h and magnetically enriched for c-kit + cells were further labelled for Sca-1, B220, PDCA-1 and electronically sorted into c-kit + Sca-1 + B220 int PDCA-1 + cells. Flow cytometry dot plots representative of 30 experiments. ( b ) Further characterization of cell-sorted CpG- as well as PBS-induced BM population was performed using flow cytometry analysis of the expression of various myeloid and dendritic cell markers as well as cell migration receptors. Specific antibody staining is depicted with open histograms (blue line for PBS- and red line for CpG-induced cells). Positive cells are defined using FMO controls (grey histograms). Two experiments. ( c ) FACS analysis of intranuclear expression of transcription factors. Analysis of IRF8 and Id2 expression with specific antibodies compared to FMO controls and of E2.2 and PU.1 by flowRNA, compared with B220 + PDCA-1 − control B cells (grey histograms), in CpG- versus PBS-stimulated c-kit + Sca-1 + B220 int PDCA-1 + population. Two experiments. ( d ) Frequency of c-kit + Sca-1 + B220 int PDCA-1 + cells emerging among total BM cells after 18 h of incubation with different TLR agonists, measured in BM cell cultures of four individual mice. Statistical significance as indicated, analyzed by Mann–Whitney test. ( e ) Cytokine production of cell-sorted PBS- versus CpG-induced c-kit + Sca-1 + B220 int PDCA-1 + cells, measured in the supernatant after 4 h of activation with PMA/ionomycin in the presence of brefeldin A using multiplex ELISA. Biological duplicates. ( f ) Intracytoplasmic TGF-β expression was analyzed by FACS in PBS- and CpG-pre-pDCs. One experiment out of two ( g ) IFN-α production was measured by ELISA in supernatants of CpG- versus PBS-induced cell-sorted c-kit + Sca-1 + B220 int PDCA-1 + BM cells after 18 h incubation with 1 μg/ml TLR-7 (R848), TLR-9 (CpG-B and CpG-P) and TLR-4 (LPS) agonists (biological duplicates). ( h ) Mature spleen pDCs and CpG-pre-pDCs were compared for their IFN-α response after 18-h incubation with CpG-B (1 μg/ml), measured by ELISA in supernatants (biological duplicates).

    Techniques Used: FACS, Functional Assay, Incubation, Flow Cytometry, Expressing, Migration, Staining, Control, MANN-WHITNEY, Activation Assay, Multiplex Assay, Enzyme-linked Immunosorbent Assay

    Role of TGF-β in CpG-pre-pDC mediated protection against EAE. ( a ) CpG-pre-pDCs were transfected with a neutralizing anti-TGF-β antibody or control isotype antibody using the Chariot protein transfection vector. 80,000 of resulting cells were injected i.v. to MOG 35-55 immunized mice at d-12 after immunization. Clinical score (mean ± s.e.m.) was assessed until d-27, n = 10 mice in control and isotype antibody progenitor-treated groups and n = 9 mice in the anti-TGF-β-progenitor-treated group. Statistical analysis was performed using two-way ANOVA with Bonferroni post-test: controls vs CpG-prepDC + isotype, d-19, * p = 0.0352; controls vs CpG-prep-DC + anti-TGF-β, d-19, *, p = 0.0144, d20–d22, *** p < 0.0025, d27 *, p = 0.0352; CpG-pre-pDCs + isotype vs CpG-pre-pDCs + anti-TGF-β, d17–18, *, p = 0.0119, d19–d27, ****, p < 0.0001. ( b ) CD4 + T-cells isolated at d-27 from the spinal cord of controls or recipients of isotype or neutralizing Ab-transfected CpG-pre-pDCs were analyzed by FACS using FMO controls for their cytokine production after 4 h activation with PMA/ionomycin in presence of brefeldin. Mean ± s.e.m. of percentages of cells expressing a given cytokine, n = 5 mice per group. Statistical analysis was performed using two-way ANOVA with Bonferroni post-tests, for GM-CSF: Controls vs CpG-pre-pDCs + isotype Ab, NS, Controls vs CpG-pre-pDCs + anti-TGF-β, **, p = 0.0032, CpG-pre-pDCs + isotype vs + anti-TGF-β, **, p = 0.008, *; for TNF-α, CpG-pre-pDCs + isotype vs + anti-TGF-β, **, p = 0.0015. ( c , d ) Frequency ( c ) and counts ( d ) of B220 + PDCA-1 − B-cells demonstrating IL-10 production capacity (mean ± s.e.m., n = 5 mice per group, * p = 0.01, **, p = 0.0015, by one-way ANOVA with Bonferroni post-tests). ( e , f ) CD4 + Foxp3 + Tregs frequency ( e ) and cell counts ( f ), (mean ± s.e.m., n = 5 mice per group), N.S. by analysis using Kruskal–Wallis with Dunn’s post-tests.
    Figure Legend Snippet: Role of TGF-β in CpG-pre-pDC mediated protection against EAE. ( a ) CpG-pre-pDCs were transfected with a neutralizing anti-TGF-β antibody or control isotype antibody using the Chariot protein transfection vector. 80,000 of resulting cells were injected i.v. to MOG 35-55 immunized mice at d-12 after immunization. Clinical score (mean ± s.e.m.) was assessed until d-27, n = 10 mice in control and isotype antibody progenitor-treated groups and n = 9 mice in the anti-TGF-β-progenitor-treated group. Statistical analysis was performed using two-way ANOVA with Bonferroni post-test: controls vs CpG-prepDC + isotype, d-19, * p = 0.0352; controls vs CpG-prep-DC + anti-TGF-β, d-19, *, p = 0.0144, d20–d22, *** p < 0.0025, d27 *, p = 0.0352; CpG-pre-pDCs + isotype vs CpG-pre-pDCs + anti-TGF-β, d17–18, *, p = 0.0119, d19–d27, ****, p < 0.0001. ( b ) CD4 + T-cells isolated at d-27 from the spinal cord of controls or recipients of isotype or neutralizing Ab-transfected CpG-pre-pDCs were analyzed by FACS using FMO controls for their cytokine production after 4 h activation with PMA/ionomycin in presence of brefeldin. Mean ± s.e.m. of percentages of cells expressing a given cytokine, n = 5 mice per group. Statistical analysis was performed using two-way ANOVA with Bonferroni post-tests, for GM-CSF: Controls vs CpG-pre-pDCs + isotype Ab, NS, Controls vs CpG-pre-pDCs + anti-TGF-β, **, p = 0.0032, CpG-pre-pDCs + isotype vs + anti-TGF-β, **, p = 0.008, *; for TNF-α, CpG-pre-pDCs + isotype vs + anti-TGF-β, **, p = 0.0015. ( c , d ) Frequency ( c ) and counts ( d ) of B220 + PDCA-1 − B-cells demonstrating IL-10 production capacity (mean ± s.e.m., n = 5 mice per group, * p = 0.01, **, p = 0.0015, by one-way ANOVA with Bonferroni post-tests). ( e , f ) CD4 + Foxp3 + Tregs frequency ( e ) and cell counts ( f ), (mean ± s.e.m., n = 5 mice per group), N.S. by analysis using Kruskal–Wallis with Dunn’s post-tests.

    Techniques Used: Transfection, Control, Plasmid Preparation, Injection, Isolation, Activation Assay, Expressing

    Graphical representation of cellular and molecular mechanisms of the protective effect of CpG-pre-pDCs against EAE. Upon adoptive transfer of pDC progenitors at d-12, the onset of clinical signs, TGF-β released only by CpG-pre-pDCs in the first 3 days in the spinal cord (light blue zone) of mice protects against EAE by prompting host pDCs to release in turn TGF-β and promoting accumulation of CD11c + CD11b + cDCs and IL-10 + B cells. CpG-pre-pDCs, once migrated to the spinal cord, additionally release IL-27 that from day-22 onwards, ensures late protection against EAE. BBB blood brain barrier. The figure was created using Biorender.com.
    Figure Legend Snippet: Graphical representation of cellular and molecular mechanisms of the protective effect of CpG-pre-pDCs against EAE. Upon adoptive transfer of pDC progenitors at d-12, the onset of clinical signs, TGF-β released only by CpG-pre-pDCs in the first 3 days in the spinal cord (light blue zone) of mice protects against EAE by prompting host pDCs to release in turn TGF-β and promoting accumulation of CD11c + CD11b + cDCs and IL-10 + B cells. CpG-pre-pDCs, once migrated to the spinal cord, additionally release IL-27 that from day-22 onwards, ensures late protection against EAE. BBB blood brain barrier. The figure was created using Biorender.com.

    Techniques Used: Adoptive Transfer Assay



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    leaf purified anti-tgf-β neutralizing antibody (clone 2d11.16.8)  (Bio X Cell)
    90
    Bio X Cell leaf purified anti-tgf-β neutralizing antibody (clone 2d11.16.8)
    Cell sorting strategy, phenotypic and functional characterization of CpG-induced c-kit + Sca-1 + B220 int PDCA-1 + BM cells. ( a ) Total BM cells incubated with PBS or CpG-B (1 µg/ml) for 18 h and magnetically enriched for c-kit + cells were further labelled for Sca-1, B220, PDCA-1 and electronically sorted into c-kit + Sca-1 + B220 int PDCA-1 + cells. Flow cytometry dot plots representative of 30 experiments. ( b ) Further characterization of cell-sorted CpG- as well as PBS-induced BM population was performed using flow cytometry analysis of the expression of various myeloid and dendritic cell markers as well as cell migration receptors. Specific antibody staining is depicted with open histograms (blue line for PBS- and red line for CpG-induced cells). Positive cells are defined using FMO controls (grey histograms). Two experiments. ( c ) FACS analysis of intranuclear expression of transcription factors. Analysis of IRF8 and Id2 expression with specific antibodies compared to FMO controls and of E2.2 and PU.1 by flowRNA, compared with B220 + PDCA-1 − control B cells (grey histograms), in CpG- versus PBS-stimulated c-kit + Sca-1 + B220 int PDCA-1 + population. Two experiments. ( d ) Frequency of c-kit + Sca-1 + B220 int PDCA-1 + cells emerging among total BM cells after 18 h of incubation with different TLR agonists, measured in BM cell cultures of four individual mice. Statistical significance as indicated, analyzed by Mann–Whitney test. ( e ) Cytokine production of cell-sorted PBS- versus CpG-induced c-kit + Sca-1 + B220 int PDCA-1 + cells, measured in the supernatant after 4 h of activation with PMA/ionomycin in the presence of brefeldin A using multiplex ELISA. Biological duplicates. ( f ) Intracytoplasmic <t>TGF-β</t> expression was analyzed by FACS in PBS- and CpG-pre-pDCs. One experiment out of two ( g ) IFN-α production was measured by ELISA in supernatants of CpG- versus PBS-induced cell-sorted c-kit + Sca-1 + B220 int PDCA-1 + BM cells after 18 h incubation with 1 μg/ml TLR-7 (R848), TLR-9 (CpG-B and CpG-P) and TLR-4 (LPS) agonists (biological duplicates). ( h ) Mature spleen pDCs and CpG-pre-pDCs were compared for their IFN-α response after 18-h incubation with CpG-B (1 μg/ml), measured by ELISA in supernatants (biological duplicates).
    Leaf Purified Anti Tgf β Neutralizing Antibody (Clone 2d11.16.8), supplied by Bio X Cell, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/leaf+purified+anti-tgf-%CE%B2+neutralizing+antibody+(clone+2d11%2E16%2E8)/leaf+purified+anti+tgf+%CE%B2+neutralizing+antibody++clone+2d11+16+8+/pmc07910458-219-15-22
    Average 90 stars, based on 1 article reviews
    leaf purified anti-tgf-β neutralizing antibody (clone 2d11.16.8) - by Bioz Stars, 2026-09
    90/100 stars
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    Cell sorting strategy, phenotypic and functional characterization of CpG-induced c-kit + Sca-1 + B220 int PDCA-1 + BM cells. ( a ) Total BM cells incubated with PBS or CpG-B (1 µg/ml) for 18 h and magnetically enriched for c-kit + cells were further labelled for Sca-1, B220, PDCA-1 and electronically sorted into c-kit + Sca-1 + B220 int PDCA-1 + cells. Flow cytometry dot plots representative of 30 experiments. ( b ) Further characterization of cell-sorted CpG- as well as PBS-induced BM population was performed using flow cytometry analysis of the expression of various myeloid and dendritic cell markers as well as cell migration receptors. Specific antibody staining is depicted with open histograms (blue line for PBS- and red line for CpG-induced cells). Positive cells are defined using FMO controls (grey histograms). Two experiments. ( c ) FACS analysis of intranuclear expression of transcription factors. Analysis of IRF8 and Id2 expression with specific antibodies compared to FMO controls and of E2.2 and PU.1 by flowRNA, compared with B220 + PDCA-1 − control B cells (grey histograms), in CpG- versus PBS-stimulated c-kit + Sca-1 + B220 int PDCA-1 + population. Two experiments. ( d ) Frequency of c-kit + Sca-1 + B220 int PDCA-1 + cells emerging among total BM cells after 18 h of incubation with different TLR agonists, measured in BM cell cultures of four individual mice. Statistical significance as indicated, analyzed by Mann–Whitney test. ( e ) Cytokine production of cell-sorted PBS- versus CpG-induced c-kit + Sca-1 + B220 int PDCA-1 + cells, measured in the supernatant after 4 h of activation with PMA/ionomycin in the presence of brefeldin A using multiplex ELISA. Biological duplicates. ( f ) Intracytoplasmic TGF-β expression was analyzed by FACS in PBS- and CpG-pre-pDCs. One experiment out of two ( g ) IFN-α production was measured by ELISA in supernatants of CpG- versus PBS-induced cell-sorted c-kit + Sca-1 + B220 int PDCA-1 + BM cells after 18 h incubation with 1 μg/ml TLR-7 (R848), TLR-9 (CpG-B and CpG-P) and TLR-4 (LPS) agonists (biological duplicates). ( h ) Mature spleen pDCs and CpG-pre-pDCs were compared for their IFN-α response after 18-h incubation with CpG-B (1 μg/ml), measured by ELISA in supernatants (biological duplicates).

    Journal: Scientific Reports

    Article Title: Toll-like receptor-9 stimulated plasmacytoid dendritic cell precursors suppress autoimmune neuroinflammation in a murine model of multiple sclerosis

    doi: 10.1038/s41598-021-84023-0

    Figure Lengend Snippet: Cell sorting strategy, phenotypic and functional characterization of CpG-induced c-kit + Sca-1 + B220 int PDCA-1 + BM cells. ( a ) Total BM cells incubated with PBS or CpG-B (1 µg/ml) for 18 h and magnetically enriched for c-kit + cells were further labelled for Sca-1, B220, PDCA-1 and electronically sorted into c-kit + Sca-1 + B220 int PDCA-1 + cells. Flow cytometry dot plots representative of 30 experiments. ( b ) Further characterization of cell-sorted CpG- as well as PBS-induced BM population was performed using flow cytometry analysis of the expression of various myeloid and dendritic cell markers as well as cell migration receptors. Specific antibody staining is depicted with open histograms (blue line for PBS- and red line for CpG-induced cells). Positive cells are defined using FMO controls (grey histograms). Two experiments. ( c ) FACS analysis of intranuclear expression of transcription factors. Analysis of IRF8 and Id2 expression with specific antibodies compared to FMO controls and of E2.2 and PU.1 by flowRNA, compared with B220 + PDCA-1 − control B cells (grey histograms), in CpG- versus PBS-stimulated c-kit + Sca-1 + B220 int PDCA-1 + population. Two experiments. ( d ) Frequency of c-kit + Sca-1 + B220 int PDCA-1 + cells emerging among total BM cells after 18 h of incubation with different TLR agonists, measured in BM cell cultures of four individual mice. Statistical significance as indicated, analyzed by Mann–Whitney test. ( e ) Cytokine production of cell-sorted PBS- versus CpG-induced c-kit + Sca-1 + B220 int PDCA-1 + cells, measured in the supernatant after 4 h of activation with PMA/ionomycin in the presence of brefeldin A using multiplex ELISA. Biological duplicates. ( f ) Intracytoplasmic TGF-β expression was analyzed by FACS in PBS- and CpG-pre-pDCs. One experiment out of two ( g ) IFN-α production was measured by ELISA in supernatants of CpG- versus PBS-induced cell-sorted c-kit + Sca-1 + B220 int PDCA-1 + BM cells after 18 h incubation with 1 μg/ml TLR-7 (R848), TLR-9 (CpG-B and CpG-P) and TLR-4 (LPS) agonists (biological duplicates). ( h ) Mature spleen pDCs and CpG-pre-pDCs were compared for their IFN-α response after 18-h incubation with CpG-B (1 μg/ml), measured by ELISA in supernatants (biological duplicates).

    Article Snippet: 0.1 mg Chariot (Active Motif, La Hupe, Belgium) was combined with 50 μg of either LEAF purified anti-TGF-β neutralizing antibody (clone 2D11.16.8) (BioXCell, West Lebanon, NH) or IgG1 isotype control (Biolegend) according to the manufacturer’s instructions.

    Techniques: FACS, Functional Assay, Incubation, Flow Cytometry, Expressing, Migration, Staining, Control, MANN-WHITNEY, Activation Assay, Multiplex Assay, Enzyme-linked Immunosorbent Assay

    Role of TGF-β in CpG-pre-pDC mediated protection against EAE. ( a ) CpG-pre-pDCs were transfected with a neutralizing anti-TGF-β antibody or control isotype antibody using the Chariot protein transfection vector. 80,000 of resulting cells were injected i.v. to MOG 35-55 immunized mice at d-12 after immunization. Clinical score (mean ± s.e.m.) was assessed until d-27, n = 10 mice in control and isotype antibody progenitor-treated groups and n = 9 mice in the anti-TGF-β-progenitor-treated group. Statistical analysis was performed using two-way ANOVA with Bonferroni post-test: controls vs CpG-prepDC + isotype, d-19, * p = 0.0352; controls vs CpG-prep-DC + anti-TGF-β, d-19, *, p = 0.0144, d20–d22, *** p < 0.0025, d27 *, p = 0.0352; CpG-pre-pDCs + isotype vs CpG-pre-pDCs + anti-TGF-β, d17–18, *, p = 0.0119, d19–d27, ****, p < 0.0001. ( b ) CD4 + T-cells isolated at d-27 from the spinal cord of controls or recipients of isotype or neutralizing Ab-transfected CpG-pre-pDCs were analyzed by FACS using FMO controls for their cytokine production after 4 h activation with PMA/ionomycin in presence of brefeldin. Mean ± s.e.m. of percentages of cells expressing a given cytokine, n = 5 mice per group. Statistical analysis was performed using two-way ANOVA with Bonferroni post-tests, for GM-CSF: Controls vs CpG-pre-pDCs + isotype Ab, NS, Controls vs CpG-pre-pDCs + anti-TGF-β, **, p = 0.0032, CpG-pre-pDCs + isotype vs + anti-TGF-β, **, p = 0.008, *; for TNF-α, CpG-pre-pDCs + isotype vs + anti-TGF-β, **, p = 0.0015. ( c , d ) Frequency ( c ) and counts ( d ) of B220 + PDCA-1 − B-cells demonstrating IL-10 production capacity (mean ± s.e.m., n = 5 mice per group, * p = 0.01, **, p = 0.0015, by one-way ANOVA with Bonferroni post-tests). ( e , f ) CD4 + Foxp3 + Tregs frequency ( e ) and cell counts ( f ), (mean ± s.e.m., n = 5 mice per group), N.S. by analysis using Kruskal–Wallis with Dunn’s post-tests.

    Journal: Scientific Reports

    Article Title: Toll-like receptor-9 stimulated plasmacytoid dendritic cell precursors suppress autoimmune neuroinflammation in a murine model of multiple sclerosis

    doi: 10.1038/s41598-021-84023-0

    Figure Lengend Snippet: Role of TGF-β in CpG-pre-pDC mediated protection against EAE. ( a ) CpG-pre-pDCs were transfected with a neutralizing anti-TGF-β antibody or control isotype antibody using the Chariot protein transfection vector. 80,000 of resulting cells were injected i.v. to MOG 35-55 immunized mice at d-12 after immunization. Clinical score (mean ± s.e.m.) was assessed until d-27, n = 10 mice in control and isotype antibody progenitor-treated groups and n = 9 mice in the anti-TGF-β-progenitor-treated group. Statistical analysis was performed using two-way ANOVA with Bonferroni post-test: controls vs CpG-prepDC + isotype, d-19, * p = 0.0352; controls vs CpG-prep-DC + anti-TGF-β, d-19, *, p = 0.0144, d20–d22, *** p < 0.0025, d27 *, p = 0.0352; CpG-pre-pDCs + isotype vs CpG-pre-pDCs + anti-TGF-β, d17–18, *, p = 0.0119, d19–d27, ****, p < 0.0001. ( b ) CD4 + T-cells isolated at d-27 from the spinal cord of controls or recipients of isotype or neutralizing Ab-transfected CpG-pre-pDCs were analyzed by FACS using FMO controls for their cytokine production after 4 h activation with PMA/ionomycin in presence of brefeldin. Mean ± s.e.m. of percentages of cells expressing a given cytokine, n = 5 mice per group. Statistical analysis was performed using two-way ANOVA with Bonferroni post-tests, for GM-CSF: Controls vs CpG-pre-pDCs + isotype Ab, NS, Controls vs CpG-pre-pDCs + anti-TGF-β, **, p = 0.0032, CpG-pre-pDCs + isotype vs + anti-TGF-β, **, p = 0.008, *; for TNF-α, CpG-pre-pDCs + isotype vs + anti-TGF-β, **, p = 0.0015. ( c , d ) Frequency ( c ) and counts ( d ) of B220 + PDCA-1 − B-cells demonstrating IL-10 production capacity (mean ± s.e.m., n = 5 mice per group, * p = 0.01, **, p = 0.0015, by one-way ANOVA with Bonferroni post-tests). ( e , f ) CD4 + Foxp3 + Tregs frequency ( e ) and cell counts ( f ), (mean ± s.e.m., n = 5 mice per group), N.S. by analysis using Kruskal–Wallis with Dunn’s post-tests.

    Article Snippet: 0.1 mg Chariot (Active Motif, La Hupe, Belgium) was combined with 50 μg of either LEAF purified anti-TGF-β neutralizing antibody (clone 2D11.16.8) (BioXCell, West Lebanon, NH) or IgG1 isotype control (Biolegend) according to the manufacturer’s instructions.

    Techniques: Transfection, Control, Plasmid Preparation, Injection, Isolation, Activation Assay, Expressing

    Graphical representation of cellular and molecular mechanisms of the protective effect of CpG-pre-pDCs against EAE. Upon adoptive transfer of pDC progenitors at d-12, the onset of clinical signs, TGF-β released only by CpG-pre-pDCs in the first 3 days in the spinal cord (light blue zone) of mice protects against EAE by prompting host pDCs to release in turn TGF-β and promoting accumulation of CD11c + CD11b + cDCs and IL-10 + B cells. CpG-pre-pDCs, once migrated to the spinal cord, additionally release IL-27 that from day-22 onwards, ensures late protection against EAE. BBB blood brain barrier. The figure was created using Biorender.com.

    Journal: Scientific Reports

    Article Title: Toll-like receptor-9 stimulated plasmacytoid dendritic cell precursors suppress autoimmune neuroinflammation in a murine model of multiple sclerosis

    doi: 10.1038/s41598-021-84023-0

    Figure Lengend Snippet: Graphical representation of cellular and molecular mechanisms of the protective effect of CpG-pre-pDCs against EAE. Upon adoptive transfer of pDC progenitors at d-12, the onset of clinical signs, TGF-β released only by CpG-pre-pDCs in the first 3 days in the spinal cord (light blue zone) of mice protects against EAE by prompting host pDCs to release in turn TGF-β and promoting accumulation of CD11c + CD11b + cDCs and IL-10 + B cells. CpG-pre-pDCs, once migrated to the spinal cord, additionally release IL-27 that from day-22 onwards, ensures late protection against EAE. BBB blood brain barrier. The figure was created using Biorender.com.

    Article Snippet: 0.1 mg Chariot (Active Motif, La Hupe, Belgium) was combined with 50 μg of either LEAF purified anti-TGF-β neutralizing antibody (clone 2D11.16.8) (BioXCell, West Lebanon, NH) or IgG1 isotype control (Biolegend) according to the manufacturer’s instructions.

    Techniques: Adoptive Transfer Assay