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cd21  (Miltenyi Biotec)


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    Structured Review

    Miltenyi Biotec cd21
    a , Counts of peripheral B cells in individuals with CBL -LOH over time compared with the healthy range. Data from eight participants are shown. Healthy ranges are from pediatric clinical recommendations. b , IgG levels in individuals with CBL -LOH (P1–P8) over time compared with the healthy range. Data from eight participants are shown. Healthy ranges are from pediatric clinical recommendations. c , Quantification of the indicated B cell subsets in the peripheral cryopreserved mononuclear cell blood of HDs, heterozygous HDs and individuals with CBL -LOH of the indicated ages as determined by mass cytometry; controls 0–3 years old ( n = 2), controls 4–15 years old ( n = 9), controls 16–100 years old ( n = 28), pediatric participants (LOH) ( n = 5), adult participants (LOH) ( n = 2), heterozygous individuals ( n = 3). Data are shown as mean ± s.d. The statistical significance of differences was assessed in multiple two-sided Mann–Whitney tests, with correction for multiple testing; ** P < 0.005 and *** P < 0.0005. y.o., years old. d , Frequency of B cell subsets in cryopreserved PBMCs from HDs and individuals with CBL -LOH as determined by flow cytometry; HDs ( n = 13), individuals with CBL -LOH ( n = 5). Data are shown as mean ± s.d. The statistical significance of differences was assessed using multiple two-sided Mann–Whitney tests, with correction for multiple testing; * P < 0.05 and ** P < 0.005. e , CD5, CD9, <t>CD21</t> and CD38 expression on transitional B cells of HDs (black) and individuals with CBL -LOH (red) as determined by flow cytometry; HDs ( n = 19), individuals with CBL -LOH ( n = 6). Data are shown as mean ± s.d. The statistical significance of differences was assessed by multiple two-sided Mann–Whitney tests, with correction for multiple testing; * P < 0.05, ** P < 0.005 and **** P < 0.00005. f , Percentage of CD21 hi cells among transitional B cells of HDs (black) and individuals with CBL -LOH (red) as determined by flow cytometry; HDs ( n = 9), individuals with CBL -LOH ( n = 5). Data are shown as mean ± s.d. The statistical significance of differences was assessed by Mann–Whitney test; *** P < 0.0005; gMFI, geometric mean fluorescence intensity.
    Cd21, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 94/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/anti+human+cd21/CD21+Antibody%2C+anti-human%2C+REAfinity/pmc12864045-23-2-8
    Average 94 stars, based on 10 article reviews
    cd21 - by Bioz Stars, 2026-09
    94/100 stars

    Images

    1) Product Images from "Somatic deficiency of the human E3 ubiquitin ligase CBL in leukocytes impairs B cell but not T cell development and function"

    Article Title: Somatic deficiency of the human E3 ubiquitin ligase CBL in leukocytes impairs B cell but not T cell development and function

    Journal: Nature Immunology

    doi: 10.1038/s41590-025-02381-7

    a , Counts of peripheral B cells in individuals with CBL -LOH over time compared with the healthy range. Data from eight participants are shown. Healthy ranges are from pediatric clinical recommendations. b , IgG levels in individuals with CBL -LOH (P1–P8) over time compared with the healthy range. Data from eight participants are shown. Healthy ranges are from pediatric clinical recommendations. c , Quantification of the indicated B cell subsets in the peripheral cryopreserved mononuclear cell blood of HDs, heterozygous HDs and individuals with CBL -LOH of the indicated ages as determined by mass cytometry; controls 0–3 years old ( n = 2), controls 4–15 years old ( n = 9), controls 16–100 years old ( n = 28), pediatric participants (LOH) ( n = 5), adult participants (LOH) ( n = 2), heterozygous individuals ( n = 3). Data are shown as mean ± s.d. The statistical significance of differences was assessed in multiple two-sided Mann–Whitney tests, with correction for multiple testing; ** P < 0.005 and *** P < 0.0005. y.o., years old. d , Frequency of B cell subsets in cryopreserved PBMCs from HDs and individuals with CBL -LOH as determined by flow cytometry; HDs ( n = 13), individuals with CBL -LOH ( n = 5). Data are shown as mean ± s.d. The statistical significance of differences was assessed using multiple two-sided Mann–Whitney tests, with correction for multiple testing; * P < 0.05 and ** P < 0.005. e , CD5, CD9, CD21 and CD38 expression on transitional B cells of HDs (black) and individuals with CBL -LOH (red) as determined by flow cytometry; HDs ( n = 19), individuals with CBL -LOH ( n = 6). Data are shown as mean ± s.d. The statistical significance of differences was assessed by multiple two-sided Mann–Whitney tests, with correction for multiple testing; * P < 0.05, ** P < 0.005 and **** P < 0.00005. f , Percentage of CD21 hi cells among transitional B cells of HDs (black) and individuals with CBL -LOH (red) as determined by flow cytometry; HDs ( n = 9), individuals with CBL -LOH ( n = 5). Data are shown as mean ± s.d. The statistical significance of differences was assessed by Mann–Whitney test; *** P < 0.0005; gMFI, geometric mean fluorescence intensity.
    Figure Legend Snippet: a , Counts of peripheral B cells in individuals with CBL -LOH over time compared with the healthy range. Data from eight participants are shown. Healthy ranges are from pediatric clinical recommendations. b , IgG levels in individuals with CBL -LOH (P1–P8) over time compared with the healthy range. Data from eight participants are shown. Healthy ranges are from pediatric clinical recommendations. c , Quantification of the indicated B cell subsets in the peripheral cryopreserved mononuclear cell blood of HDs, heterozygous HDs and individuals with CBL -LOH of the indicated ages as determined by mass cytometry; controls 0–3 years old ( n = 2), controls 4–15 years old ( n = 9), controls 16–100 years old ( n = 28), pediatric participants (LOH) ( n = 5), adult participants (LOH) ( n = 2), heterozygous individuals ( n = 3). Data are shown as mean ± s.d. The statistical significance of differences was assessed in multiple two-sided Mann–Whitney tests, with correction for multiple testing; ** P < 0.005 and *** P < 0.0005. y.o., years old. d , Frequency of B cell subsets in cryopreserved PBMCs from HDs and individuals with CBL -LOH as determined by flow cytometry; HDs ( n = 13), individuals with CBL -LOH ( n = 5). Data are shown as mean ± s.d. The statistical significance of differences was assessed using multiple two-sided Mann–Whitney tests, with correction for multiple testing; * P < 0.05 and ** P < 0.005. e , CD5, CD9, CD21 and CD38 expression on transitional B cells of HDs (black) and individuals with CBL -LOH (red) as determined by flow cytometry; HDs ( n = 19), individuals with CBL -LOH ( n = 6). Data are shown as mean ± s.d. The statistical significance of differences was assessed by multiple two-sided Mann–Whitney tests, with correction for multiple testing; * P < 0.05, ** P < 0.005 and **** P < 0.00005. f , Percentage of CD21 hi cells among transitional B cells of HDs (black) and individuals with CBL -LOH (red) as determined by flow cytometry; HDs ( n = 9), individuals with CBL -LOH ( n = 5). Data are shown as mean ± s.d. The statistical significance of differences was assessed by Mann–Whitney test; *** P < 0.0005; gMFI, geometric mean fluorescence intensity.

    Techniques Used: Mass Cytometry, MANN-WHITNEY, Flow Cytometry, Expressing, Fluorescence

    ( a ) Quantification of B-cell subsets in cryopreserved PBMCs from healthy donors, heterozygous carriers, and CBL-LOH patients of the indicated ages by mass cytometry. Controls 0–3 y.o. (n = 2); controls 4–15 y.o. (n = 9); controls 16–100 y.o. (n = 28); pediatric CBL-LOH (n = 5); adult CBL-LOH (n = 2); heterozygous individuals (n = 3). Mean ± s.d. Statistical significance was assessed using two-sided Mann–Whitney tests with correction for multiple testing. ( b ) Frequency of B-cell subsets in healthy donors (n = 10) and CBL-LOH patients (n = 4) by flow cytometry. Mean ± s.d. Significance was evaluated with two-sided Mann–Whitney tests adjusted for multiple comparisons. **p < 0.005. ( c ) MFI of B-cell markers on CD21 lo and transitional B cells from healthy donors (n = 6) and patients P1–3 (n = 3). Lines show means. ( d–f ) Modeling the CBL ΔExon8 variant in primary human CD34⁺ HSPCs. ( d ) Agarose gel electrophoresis of PCR products from AAVS1 or CBL loci 72 h after nucleofection; representative of >5 biological replicates. ( e ) Editing efficiency using sgRNA pair 1+2 by NGS, showing ~80% of the ~400 bp exon-8 deletion. ( f ) Western blot of CBL protein levels after CBL or AAVS1 editing. ( g ) Quantification of three biological replicates from (f). Mean ± s.d. ( h ) NGS quantification of exon-8 deletions for all three guide pairs (n = 2 biological replicates). Bars show means. ( i ) Editing efficiencies at days 8 and 21 in differentiation cultures by NGS. Mean ± s.d. from three biological replicates. ( j–m ) Bulk RNA-seq of AAVS1- and CBL-edited CD19⁺CD10⁺CD20 low HSPC-derived B-cell progenitors. ( j ) Gene-set enrichment analysis showing significantly enriched (red) or depleted (blue) pathways (NES: normalized enrichment score). ( k,l ) Differential expression of leading-edge genes in the ( k ) Hallmark G2–M checkpoint and ( l ) Hallmark mTORC1 signaling gene sets (Z-transformed normalized counts). ( m ) Transcriptional overlap between CBL-edited and PI3K GOF progenitors; Venn diagrams show shared significantly up- or downregulated genes, with overlap significance by binomial test.
    Figure Legend Snippet: ( a ) Quantification of B-cell subsets in cryopreserved PBMCs from healthy donors, heterozygous carriers, and CBL-LOH patients of the indicated ages by mass cytometry. Controls 0–3 y.o. (n = 2); controls 4–15 y.o. (n = 9); controls 16–100 y.o. (n = 28); pediatric CBL-LOH (n = 5); adult CBL-LOH (n = 2); heterozygous individuals (n = 3). Mean ± s.d. Statistical significance was assessed using two-sided Mann–Whitney tests with correction for multiple testing. ( b ) Frequency of B-cell subsets in healthy donors (n = 10) and CBL-LOH patients (n = 4) by flow cytometry. Mean ± s.d. Significance was evaluated with two-sided Mann–Whitney tests adjusted for multiple comparisons. **p < 0.005. ( c ) MFI of B-cell markers on CD21 lo and transitional B cells from healthy donors (n = 6) and patients P1–3 (n = 3). Lines show means. ( d–f ) Modeling the CBL ΔExon8 variant in primary human CD34⁺ HSPCs. ( d ) Agarose gel electrophoresis of PCR products from AAVS1 or CBL loci 72 h after nucleofection; representative of >5 biological replicates. ( e ) Editing efficiency using sgRNA pair 1+2 by NGS, showing ~80% of the ~400 bp exon-8 deletion. ( f ) Western blot of CBL protein levels after CBL or AAVS1 editing. ( g ) Quantification of three biological replicates from (f). Mean ± s.d. ( h ) NGS quantification of exon-8 deletions for all three guide pairs (n = 2 biological replicates). Bars show means. ( i ) Editing efficiencies at days 8 and 21 in differentiation cultures by NGS. Mean ± s.d. from three biological replicates. ( j–m ) Bulk RNA-seq of AAVS1- and CBL-edited CD19⁺CD10⁺CD20 low HSPC-derived B-cell progenitors. ( j ) Gene-set enrichment analysis showing significantly enriched (red) or depleted (blue) pathways (NES: normalized enrichment score). ( k,l ) Differential expression of leading-edge genes in the ( k ) Hallmark G2–M checkpoint and ( l ) Hallmark mTORC1 signaling gene sets (Z-transformed normalized counts). ( m ) Transcriptional overlap between CBL-edited and PI3K GOF progenitors; Venn diagrams show shared significantly up- or downregulated genes, with overlap significance by binomial test.

    Techniques Used: Mass Cytometry, MANN-WHITNEY, Flow Cytometry, Variant Assay, Agarose Gel Electrophoresis, Western Blot, RNA Sequencing, Derivative Assay, Quantitative Proteomics, Transformation Assay

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    Labeling:

    Article Title: Activation-Induced FoxP3 Expression Regulates Cytokine Production in Conventional T Cells Stimulated with Autologous Dendritic Cells
    Article Snippet: For negative selection of peripheral blood T cells, monocytes and neutrophils were depleted with anti-canine CD172a (hybridoma clone SWC3, VMRD, Pullman, WA) ( 22 ), and B cells were depleted with anti-human CD21 (hybridoma clone B-ly4; Becton Dickinson) ( 33 ). .. A total of 10 8 PBMC were labeled with anti-canine CD172a and anti-human CD21, washed in PBS, incubated with anti-mouse IgG1 microbeads, and eluted from an LD column according to the manufacturer's protocol (Miltenyi Biotech). .. The purified T cells were washed in PBS, resuspended in 0.25 μM CFSE (Sigma-Aldrich, St. Louis, MO) diluted in PBS, and incubated on a spinning rack at room temperature in the dark for 10 min ( 14 ).

    Article Title: Activation-Induced FoxP3 Expression Regulates Cytokine Production in Conventional T Cells Stimulated with Autologous Dendritic Cells
    Article Snippet: For negative selection of peripheral blood 111 T cells, monocytes and neutrophils were depleted with anti-canine CD172a (hybridoma clone 112 SWC3, VMRD, Pullman, WA) (22) and B cells were depleted with anti-human CD21 113 (hybridoma clone B-ly4, Becton Dickinson) (33). .. 1 x 108 PBMCs were labeled with anti-canine 114 CD172a and anti-human CD21, washed in PBS, incubated with anti-mouse IgG1 microbeads, 115 and eluted from an LD column according to the manufacturer’s protocol (Miltenyi Biotech). .. The 116 purified T cells were washed in PBS, resuspended in 0.25 μM CFSE (Sigma-Aldrich, St. Louis, 117 MO) diluted in PBS, and incubated on a spinning rack at room temperature in the dark for 10 118 min (14).

    Incubation:

    Article Title: Activation-Induced FoxP3 Expression Regulates Cytokine Production in Conventional T Cells Stimulated with Autologous Dendritic Cells
    Article Snippet: For negative selection of peripheral blood T cells, monocytes and neutrophils were depleted with anti-canine CD172a (hybridoma clone SWC3, VMRD, Pullman, WA) ( 22 ), and B cells were depleted with anti-human CD21 (hybridoma clone B-ly4; Becton Dickinson) ( 33 ). .. A total of 10 8 PBMC were labeled with anti-canine CD172a and anti-human CD21, washed in PBS, incubated with anti-mouse IgG1 microbeads, and eluted from an LD column according to the manufacturer's protocol (Miltenyi Biotech). .. The purified T cells were washed in PBS, resuspended in 0.25 μM CFSE (Sigma-Aldrich, St. Louis, MO) diluted in PBS, and incubated on a spinning rack at room temperature in the dark for 10 min ( 14 ).

    Article Title: Brucella canis induces canine CD4 + T cells multi-cytokine Th1/Th17 production via dendritic cell activation.
    Article Snippet: The naïve CD4+ T Cell Isolation Kit (MACS, Miltenyi Biotec, Bergich Gladbach, Germany) was used for human samples. .. Canine cells were incubated with anti-canine CD11b (clone CA16.3E10) and anti-human CD21 (clone LT21) previously shown to react with canine CD21 [12] (Invitrogen, Carlsbad, CA, USA), followed by Anti-Mouse IgG MicroBeads (MACS, Miltenyi Biotec, Bergich Gladbach, Germany). ..

    Article Title: Activation-Induced FoxP3 Expression Regulates Cytokine Production in Conventional T Cells Stimulated with Autologous Dendritic Cells
    Article Snippet: For negative selection of peripheral blood 111 T cells, monocytes and neutrophils were depleted with anti-canine CD172a (hybridoma clone 112 SWC3, VMRD, Pullman, WA) (22) and B cells were depleted with anti-human CD21 113 (hybridoma clone B-ly4, Becton Dickinson) (33). .. 1 x 108 PBMCs were labeled with anti-canine 114 CD172a and anti-human CD21, washed in PBS, incubated with anti-mouse IgG1 microbeads, 115 and eluted from an LD column according to the manufacturer’s protocol (Miltenyi Biotech). .. The 116 purified T cells were washed in PBS, resuspended in 0.25 μM CFSE (Sigma-Aldrich, St. Louis, 117 MO) diluted in PBS, and incubated on a spinning rack at room temperature in the dark for 10 118 min (14).

    Magnetic Cell Separation:

    Article Title: Brucella canis induces canine CD4 + T cells multi-cytokine Th1/Th17 production via dendritic cell activation.
    Article Snippet: The naïve CD4+ T Cell Isolation Kit (MACS, Miltenyi Biotec, Bergich Gladbach, Germany) was used for human samples. .. Canine cells were incubated with anti-canine CD11b (clone CA16.3E10) and anti-human CD21 (clone LT21) previously shown to react with canine CD21 [12] (Invitrogen, Carlsbad, CA, USA), followed by Anti-Mouse IgG MicroBeads (MACS, Miltenyi Biotec, Bergich Gladbach, Germany). ..



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    a , Counts of peripheral B cells in individuals with CBL -LOH over time compared with the healthy range. Data from eight participants are shown. Healthy ranges are from pediatric clinical recommendations. b , IgG levels in individuals with CBL -LOH (P1–P8) over time compared with the healthy range. Data from eight participants are shown. Healthy ranges are from pediatric clinical recommendations. c , Quantification of the indicated B cell subsets in the peripheral cryopreserved mononuclear cell blood of HDs, heterozygous HDs and individuals with CBL -LOH of the indicated ages as determined by mass cytometry; controls 0–3 years old ( n = 2), controls 4–15 years old ( n = 9), controls 16–100 years old ( n = 28), pediatric participants (LOH) ( n = 5), adult participants (LOH) ( n = 2), heterozygous individuals ( n = 3). Data are shown as mean ± s.d. The statistical significance of differences was assessed in multiple two-sided Mann–Whitney tests, with correction for multiple testing; ** P < 0.005 and *** P < 0.0005. y.o., years old. d , Frequency of B cell subsets in cryopreserved PBMCs from HDs and individuals with CBL -LOH as determined by flow cytometry; HDs ( n = 13), individuals with CBL -LOH ( n = 5). Data are shown as mean ± s.d. The statistical significance of differences was assessed using multiple two-sided Mann–Whitney tests, with correction for multiple testing; * P < 0.05 and ** P < 0.005. e , CD5, CD9, <t>CD21</t> and CD38 expression on transitional B cells of HDs (black) and individuals with CBL -LOH (red) as determined by flow cytometry; HDs ( n = 19), individuals with CBL -LOH ( n = 6). Data are shown as mean ± s.d. The statistical significance of differences was assessed by multiple two-sided Mann–Whitney tests, with correction for multiple testing; * P < 0.05, ** P < 0.005 and **** P < 0.00005. f , Percentage of CD21 hi cells among transitional B cells of HDs (black) and individuals with CBL -LOH (red) as determined by flow cytometry; HDs ( n = 9), individuals with CBL -LOH ( n = 5). Data are shown as mean ± s.d. The statistical significance of differences was assessed by Mann–Whitney test; *** P < 0.0005; gMFI, geometric mean fluorescence intensity.
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    a , Counts of peripheral B cells in individuals with CBL -LOH over time compared with the healthy range. Data from eight participants are shown. Healthy ranges are from pediatric clinical recommendations. b , IgG levels in individuals with CBL -LOH (P1–P8) over time compared with the healthy range. Data from eight participants are shown. Healthy ranges are from pediatric clinical recommendations. c , Quantification of the indicated B cell subsets in the peripheral cryopreserved mononuclear cell blood of HDs, heterozygous HDs and individuals with CBL -LOH of the indicated ages as determined by mass cytometry; controls 0–3 years old ( n = 2), controls 4–15 years old ( n = 9), controls 16–100 years old ( n = 28), pediatric participants (LOH) ( n = 5), adult participants (LOH) ( n = 2), heterozygous individuals ( n = 3). Data are shown as mean ± s.d. The statistical significance of differences was assessed in multiple two-sided Mann–Whitney tests, with correction for multiple testing; ** P < 0.005 and *** P < 0.0005. y.o., years old. d , Frequency of B cell subsets in cryopreserved PBMCs from HDs and individuals with CBL -LOH as determined by flow cytometry; HDs ( n = 13), individuals with CBL -LOH ( n = 5). Data are shown as mean ± s.d. The statistical significance of differences was assessed using multiple two-sided Mann–Whitney tests, with correction for multiple testing; * P < 0.05 and ** P < 0.005. e , CD5, CD9, <t>CD21</t> and CD38 expression on transitional B cells of HDs (black) and individuals with CBL -LOH (red) as determined by flow cytometry; HDs ( n = 19), individuals with CBL -LOH ( n = 6). Data are shown as mean ± s.d. The statistical significance of differences was assessed by multiple two-sided Mann–Whitney tests, with correction for multiple testing; * P < 0.05, ** P < 0.005 and **** P < 0.00005. f , Percentage of CD21 hi cells among transitional B cells of HDs (black) and individuals with CBL -LOH (red) as determined by flow cytometry; HDs ( n = 9), individuals with CBL -LOH ( n = 5). Data are shown as mean ± s.d. The statistical significance of differences was assessed by Mann–Whitney test; *** P < 0.0005; gMFI, geometric mean fluorescence intensity.
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    a , Counts of peripheral B cells in individuals with CBL -LOH over time compared with the healthy range. Data from eight participants are shown. Healthy ranges are from pediatric clinical recommendations. b , IgG levels in individuals with CBL -LOH (P1–P8) over time compared with the healthy range. Data from eight participants are shown. Healthy ranges are from pediatric clinical recommendations. c , Quantification of the indicated B cell subsets in the peripheral cryopreserved mononuclear cell blood of HDs, heterozygous HDs and individuals with CBL -LOH of the indicated ages as determined by mass cytometry; controls 0–3 years old ( n = 2), controls 4–15 years old ( n = 9), controls 16–100 years old ( n = 28), pediatric participants (LOH) ( n = 5), adult participants (LOH) ( n = 2), heterozygous individuals ( n = 3). Data are shown as mean ± s.d. The statistical significance of differences was assessed in multiple two-sided Mann–Whitney tests, with correction for multiple testing; ** P < 0.005 and *** P < 0.0005. y.o., years old. d , Frequency of B cell subsets in cryopreserved PBMCs from HDs and individuals with CBL -LOH as determined by flow cytometry; HDs ( n = 13), individuals with CBL -LOH ( n = 5). Data are shown as mean ± s.d. The statistical significance of differences was assessed using multiple two-sided Mann–Whitney tests, with correction for multiple testing; * P < 0.05 and ** P < 0.005. e , CD5, CD9, <t>CD21</t> and CD38 expression on transitional B cells of HDs (black) and individuals with CBL -LOH (red) as determined by flow cytometry; HDs ( n = 19), individuals with CBL -LOH ( n = 6). Data are shown as mean ± s.d. The statistical significance of differences was assessed by multiple two-sided Mann–Whitney tests, with correction for multiple testing; * P < 0.05, ** P < 0.005 and **** P < 0.00005. f , Percentage of CD21 hi cells among transitional B cells of HDs (black) and individuals with CBL -LOH (red) as determined by flow cytometry; HDs ( n = 9), individuals with CBL -LOH ( n = 5). Data are shown as mean ± s.d. The statistical significance of differences was assessed by Mann–Whitney test; *** P < 0.0005; gMFI, geometric mean fluorescence intensity.
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    a , Counts of peripheral B cells in individuals with CBL -LOH over time compared with the healthy range. Data from eight participants are shown. Healthy ranges are from pediatric clinical recommendations. b , IgG levels in individuals with CBL -LOH (P1–P8) over time compared with the healthy range. Data from eight participants are shown. Healthy ranges are from pediatric clinical recommendations. c , Quantification of the indicated B cell subsets in the peripheral cryopreserved mononuclear cell blood of HDs, heterozygous HDs and individuals with CBL -LOH of the indicated ages as determined by mass cytometry; controls 0–3 years old ( n = 2), controls 4–15 years old ( n = 9), controls 16–100 years old ( n = 28), pediatric participants (LOH) ( n = 5), adult participants (LOH) ( n = 2), heterozygous individuals ( n = 3). Data are shown as mean ± s.d. The statistical significance of differences was assessed in multiple two-sided Mann–Whitney tests, with correction for multiple testing; ** P < 0.005 and *** P < 0.0005. y.o., years old. d , Frequency of B cell subsets in cryopreserved PBMCs from HDs and individuals with CBL -LOH as determined by flow cytometry; HDs ( n = 13), individuals with CBL -LOH ( n = 5). Data are shown as mean ± s.d. The statistical significance of differences was assessed using multiple two-sided Mann–Whitney tests, with correction for multiple testing; * P < 0.05 and ** P < 0.005. e , CD5, CD9, <t>CD21</t> and CD38 expression on transitional B cells of HDs (black) and individuals with CBL -LOH (red) as determined by flow cytometry; HDs ( n = 19), individuals with CBL -LOH ( n = 6). Data are shown as mean ± s.d. The statistical significance of differences was assessed by multiple two-sided Mann–Whitney tests, with correction for multiple testing; * P < 0.05, ** P < 0.005 and **** P < 0.00005. f , Percentage of CD21 hi cells among transitional B cells of HDs (black) and individuals with CBL -LOH (red) as determined by flow cytometry; HDs ( n = 9), individuals with CBL -LOH ( n = 5). Data are shown as mean ± s.d. The statistical significance of differences was assessed by Mann–Whitney test; *** P < 0.0005; gMFI, geometric mean fluorescence intensity.
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    Image Search Results


    a , Counts of peripheral B cells in individuals with CBL -LOH over time compared with the healthy range. Data from eight participants are shown. Healthy ranges are from pediatric clinical recommendations. b , IgG levels in individuals with CBL -LOH (P1–P8) over time compared with the healthy range. Data from eight participants are shown. Healthy ranges are from pediatric clinical recommendations. c , Quantification of the indicated B cell subsets in the peripheral cryopreserved mononuclear cell blood of HDs, heterozygous HDs and individuals with CBL -LOH of the indicated ages as determined by mass cytometry; controls 0–3 years old ( n = 2), controls 4–15 years old ( n = 9), controls 16–100 years old ( n = 28), pediatric participants (LOH) ( n = 5), adult participants (LOH) ( n = 2), heterozygous individuals ( n = 3). Data are shown as mean ± s.d. The statistical significance of differences was assessed in multiple two-sided Mann–Whitney tests, with correction for multiple testing; ** P < 0.005 and *** P < 0.0005. y.o., years old. d , Frequency of B cell subsets in cryopreserved PBMCs from HDs and individuals with CBL -LOH as determined by flow cytometry; HDs ( n = 13), individuals with CBL -LOH ( n = 5). Data are shown as mean ± s.d. The statistical significance of differences was assessed using multiple two-sided Mann–Whitney tests, with correction for multiple testing; * P < 0.05 and ** P < 0.005. e , CD5, CD9, CD21 and CD38 expression on transitional B cells of HDs (black) and individuals with CBL -LOH (red) as determined by flow cytometry; HDs ( n = 19), individuals with CBL -LOH ( n = 6). Data are shown as mean ± s.d. The statistical significance of differences was assessed by multiple two-sided Mann–Whitney tests, with correction for multiple testing; * P < 0.05, ** P < 0.005 and **** P < 0.00005. f , Percentage of CD21 hi cells among transitional B cells of HDs (black) and individuals with CBL -LOH (red) as determined by flow cytometry; HDs ( n = 9), individuals with CBL -LOH ( n = 5). Data are shown as mean ± s.d. The statistical significance of differences was assessed by Mann–Whitney test; *** P < 0.0005; gMFI, geometric mean fluorescence intensity.

    Journal: Nature Immunology

    Article Title: Somatic deficiency of the human E3 ubiquitin ligase CBL in leukocytes impairs B cell but not T cell development and function

    doi: 10.1038/s41590-025-02381-7

    Figure Lengend Snippet: a , Counts of peripheral B cells in individuals with CBL -LOH over time compared with the healthy range. Data from eight participants are shown. Healthy ranges are from pediatric clinical recommendations. b , IgG levels in individuals with CBL -LOH (P1–P8) over time compared with the healthy range. Data from eight participants are shown. Healthy ranges are from pediatric clinical recommendations. c , Quantification of the indicated B cell subsets in the peripheral cryopreserved mononuclear cell blood of HDs, heterozygous HDs and individuals with CBL -LOH of the indicated ages as determined by mass cytometry; controls 0–3 years old ( n = 2), controls 4–15 years old ( n = 9), controls 16–100 years old ( n = 28), pediatric participants (LOH) ( n = 5), adult participants (LOH) ( n = 2), heterozygous individuals ( n = 3). Data are shown as mean ± s.d. The statistical significance of differences was assessed in multiple two-sided Mann–Whitney tests, with correction for multiple testing; ** P < 0.005 and *** P < 0.0005. y.o., years old. d , Frequency of B cell subsets in cryopreserved PBMCs from HDs and individuals with CBL -LOH as determined by flow cytometry; HDs ( n = 13), individuals with CBL -LOH ( n = 5). Data are shown as mean ± s.d. The statistical significance of differences was assessed using multiple two-sided Mann–Whitney tests, with correction for multiple testing; * P < 0.05 and ** P < 0.005. e , CD5, CD9, CD21 and CD38 expression on transitional B cells of HDs (black) and individuals with CBL -LOH (red) as determined by flow cytometry; HDs ( n = 19), individuals with CBL -LOH ( n = 6). Data are shown as mean ± s.d. The statistical significance of differences was assessed by multiple two-sided Mann–Whitney tests, with correction for multiple testing; * P < 0.05, ** P < 0.005 and **** P < 0.00005. f , Percentage of CD21 hi cells among transitional B cells of HDs (black) and individuals with CBL -LOH (red) as determined by flow cytometry; HDs ( n = 9), individuals with CBL -LOH ( n = 5). Data are shown as mean ± s.d. The statistical significance of differences was assessed by Mann–Whitney test; *** P < 0.0005; gMFI, geometric mean fluorescence intensity.

    Article Snippet: 162Dy , CD21 , REA940 , 130-124-315 , Miltenyi Biotec.

    Techniques: Mass Cytometry, MANN-WHITNEY, Flow Cytometry, Expressing, Fluorescence

    ( a ) Quantification of B-cell subsets in cryopreserved PBMCs from healthy donors, heterozygous carriers, and CBL-LOH patients of the indicated ages by mass cytometry. Controls 0–3 y.o. (n = 2); controls 4–15 y.o. (n = 9); controls 16–100 y.o. (n = 28); pediatric CBL-LOH (n = 5); adult CBL-LOH (n = 2); heterozygous individuals (n = 3). Mean ± s.d. Statistical significance was assessed using two-sided Mann–Whitney tests with correction for multiple testing. ( b ) Frequency of B-cell subsets in healthy donors (n = 10) and CBL-LOH patients (n = 4) by flow cytometry. Mean ± s.d. Significance was evaluated with two-sided Mann–Whitney tests adjusted for multiple comparisons. **p < 0.005. ( c ) MFI of B-cell markers on CD21 lo and transitional B cells from healthy donors (n = 6) and patients P1–3 (n = 3). Lines show means. ( d–f ) Modeling the CBL ΔExon8 variant in primary human CD34⁺ HSPCs. ( d ) Agarose gel electrophoresis of PCR products from AAVS1 or CBL loci 72 h after nucleofection; representative of >5 biological replicates. ( e ) Editing efficiency using sgRNA pair 1+2 by NGS, showing ~80% of the ~400 bp exon-8 deletion. ( f ) Western blot of CBL protein levels after CBL or AAVS1 editing. ( g ) Quantification of three biological replicates from (f). Mean ± s.d. ( h ) NGS quantification of exon-8 deletions for all three guide pairs (n = 2 biological replicates). Bars show means. ( i ) Editing efficiencies at days 8 and 21 in differentiation cultures by NGS. Mean ± s.d. from three biological replicates. ( j–m ) Bulk RNA-seq of AAVS1- and CBL-edited CD19⁺CD10⁺CD20 low HSPC-derived B-cell progenitors. ( j ) Gene-set enrichment analysis showing significantly enriched (red) or depleted (blue) pathways (NES: normalized enrichment score). ( k,l ) Differential expression of leading-edge genes in the ( k ) Hallmark G2–M checkpoint and ( l ) Hallmark mTORC1 signaling gene sets (Z-transformed normalized counts). ( m ) Transcriptional overlap between CBL-edited and PI3K GOF progenitors; Venn diagrams show shared significantly up- or downregulated genes, with overlap significance by binomial test.

    Journal: Nature Immunology

    Article Title: Somatic deficiency of the human E3 ubiquitin ligase CBL in leukocytes impairs B cell but not T cell development and function

    doi: 10.1038/s41590-025-02381-7

    Figure Lengend Snippet: ( a ) Quantification of B-cell subsets in cryopreserved PBMCs from healthy donors, heterozygous carriers, and CBL-LOH patients of the indicated ages by mass cytometry. Controls 0–3 y.o. (n = 2); controls 4–15 y.o. (n = 9); controls 16–100 y.o. (n = 28); pediatric CBL-LOH (n = 5); adult CBL-LOH (n = 2); heterozygous individuals (n = 3). Mean ± s.d. Statistical significance was assessed using two-sided Mann–Whitney tests with correction for multiple testing. ( b ) Frequency of B-cell subsets in healthy donors (n = 10) and CBL-LOH patients (n = 4) by flow cytometry. Mean ± s.d. Significance was evaluated with two-sided Mann–Whitney tests adjusted for multiple comparisons. **p < 0.005. ( c ) MFI of B-cell markers on CD21 lo and transitional B cells from healthy donors (n = 6) and patients P1–3 (n = 3). Lines show means. ( d–f ) Modeling the CBL ΔExon8 variant in primary human CD34⁺ HSPCs. ( d ) Agarose gel electrophoresis of PCR products from AAVS1 or CBL loci 72 h after nucleofection; representative of >5 biological replicates. ( e ) Editing efficiency using sgRNA pair 1+2 by NGS, showing ~80% of the ~400 bp exon-8 deletion. ( f ) Western blot of CBL protein levels after CBL or AAVS1 editing. ( g ) Quantification of three biological replicates from (f). Mean ± s.d. ( h ) NGS quantification of exon-8 deletions for all three guide pairs (n = 2 biological replicates). Bars show means. ( i ) Editing efficiencies at days 8 and 21 in differentiation cultures by NGS. Mean ± s.d. from three biological replicates. ( j–m ) Bulk RNA-seq of AAVS1- and CBL-edited CD19⁺CD10⁺CD20 low HSPC-derived B-cell progenitors. ( j ) Gene-set enrichment analysis showing significantly enriched (red) or depleted (blue) pathways (NES: normalized enrichment score). ( k,l ) Differential expression of leading-edge genes in the ( k ) Hallmark G2–M checkpoint and ( l ) Hallmark mTORC1 signaling gene sets (Z-transformed normalized counts). ( m ) Transcriptional overlap between CBL-edited and PI3K GOF progenitors; Venn diagrams show shared significantly up- or downregulated genes, with overlap significance by binomial test.

    Article Snippet: 162Dy , CD21 , REA940 , 130-124-315 , Miltenyi Biotec.

    Techniques: Mass Cytometry, MANN-WHITNEY, Flow Cytometry, Variant Assay, Agarose Gel Electrophoresis, Western Blot, RNA Sequencing, Derivative Assay, Quantitative Proteomics, Transformation Assay