Journal: Nature Cell Biology

Article Title: A single-cell atlas of non-haematopoietic cells in human lymph nodes and lymphoma reveals a landscape of stromal remodelling

doi: 10.1038/s41556-022-00866-3

Figure Lengend Snippet: a , Strategies used to identify malignant B-cell components in FL samples in silico . Shown are representative cases with light chain kappa (FL 4; top) or lambda (FL 8; bottom) restrictions confirmed by flow cytometry analysis (data not shown). After identifying B-cell components by detecting CD79A expression, we assessed expression of IGKC (for light chain kappa) and IGLC2 (for light chain lambda). Clusters with cells expressing IGKC and those expressing IGLC2 were considered non-malignant B cells, while clusters with cells expressing only one of these genes were considered malignant B cells. b , Scatter plot showing clear discrimination of malignant (filled circles) from non-malignant (empty circles) B-cell clusters in each FL sample, based on the ratio of cells expressing IGLC2 (expression level >1; y-axis) to those expressing IGKC (expression level >2; x-axis,). Red-shaded areas indicate regions in which the ratio was >2.0 or <0.25. c , Representative UMAP plots showing B cells from FL 4 according to B-cell types (beige; non-malignant, red; malignant) (left panel) or malignant B-cell signature score (right panel). d , Violin plots showing malignant B-cell signature score in extracted non-malignant and malignant B cells, according to different FL samples (FL 2–10). *** P = 1.1 × 10 −204 (FL 2), *** P = 0 (FL 3), *** P = 3.3 × 10 −176 (FL 4), *** P = 0 (FL 5), *** P = 4.2 × 10 −122 (FL 6), *** P = 0 (FL 7), *** P = 4.2 × 10 −161 (FL 8), *** P = 3.7 × 10 −256 (FL 9), *** P = 2.5 × 10 −81 (FL 10) (two-sided Wilcoxon Rank-Sum test with Bonferroni correction). e , Violin plots showing the expression of CD27 in non-malignant and malignant B cells. *** P = 0 (two-sided Wilcoxon Rank-Sum test with Bonferroni correction). f , Comparison of CD27 mean fluorescence intensity (MFI) between FL CD19 + CD10 − (non-malignant B-cell fraction) and CD19 + CD10 + (malignant B-cell fraction) cells. Circles represent biologically independent samples ( n = 8; FL 11–18). * P = 0.039 (two-sided Wilcoxon matched-pairs signed rank test). g , Flow cytometry analysis of CD27 expression on CD19 + CD10 − and CD19 + CD10 + cells of a representative FL sample (FL 14). The statistical source data are provided.

Article Snippet: B cells were isolated from the FL haematopoietic cell suspension using an EasySep Release Human CD19 Positive Selection kit (StemCell Technologies, ST-17754).

Techniques: In Silico, Flow Cytometry, Expressing, Comparison, Fluorescence

Journal: Nature Cell Biology

Article Title: A single-cell atlas of non-haematopoietic cells in human lymph nodes and lymphoma reveals a landscape of stromal remodelling

doi: 10.1038/s41556-022-00866-3

Figure Lengend Snippet: a , Enhanced interactions across FL NHC subclusters and malignant B cells (B malignant ). Circle size indicates the negative log 10 of adjusted P values . Circles are coloured when a stroma-derived factor is upregulated in relevant FL subclusters. b , IF staining for MECA-79 (cyan), DCN (red) and CD70 (green) in MFLN and FL samples. Scale bars, 200 μm. Representative images from one of three independent experiments are shown. c , Proportions of CD70 + area in medullary and adventitia regions of MFLN ( n = 3) and FL ( n = 3) samples. Circles represent biologically independent samples. Bars indicate the median. ** P = 0.0095 (two-sided unpaired t -test). d , Binding of FL CD19 + CD10 + cells to CD70-Fc protein with an anti-CD27 blocking antibody or isotype human IgG. The histograms represent three independent experiments (FL 13) with the count in arbitrary units. e , Blocking of FL CD19 + CD10 + cell binding to CD70-Fc protein after treating cells with an anti-CD27 blocking antibody ( n = 3) or isotype mouse IgG1 ( n = 3) in CD27 + FL samples (FL 11–FL 14). Proportions of cells bound to CD70-Fc protein were adjusted by subtracting nonspecific binding observed with human IgG. CD70-Fc protein binding to cells treated with isotype mouse IgG1 was set to 100% in each experiment. Circles represent independent experiments. Bars indicate the median. ** P = 0.0022, *** P = 7.3 × 10 −4 (FL 11), *** P = 2.2 × 10 −4 (FL 12), *** P = 7.6 × 10 −4 (FL 13) (two-sided paired t -test). f , Representative malignant B-enriched cell (FL 14) adhesion to medullary regions of FL in the presence of an isotype mouse IgG1 or anti-CD27 antibody. Orange dots indicate adherent cells. Yellow dashed lines indicate medullary regions. Scale bars, 200 μm. g , Blocking of malignant B-enriched cell (FL 11, FL 13 and FL 14) adhesion to FL medullary regions (per mm 2 ) after treating cells with an anti-CD27 blocking antibody ( n = 3) or isotype mouse IgG1 ( n = 3). Adhesion of cells treated with isotype mouse IgG1 was set to 100% in each experiment. Circles represent independent experiments. Bars indicate the median. * P = 0.041 (FL 11), * P = 0.027 (FL 14), ** P = 0.0050 (two-sided paired t -test). Statistical source data are provided.

Article Snippet: B cells were isolated from the FL haematopoietic cell suspension using an EasySep Release Human CD19 Positive Selection kit (StemCell Technologies, ST-17754).

Techniques: Derivative Assay, Staining, Binding Assay, Blocking Assay, Protein Binding

Journal: Mucosal immunology

Article Title: Regulatory B Cells from Hilar Lymph Nodes of Tolerant Mice in a Murine Model of Allergic Airway Disease are CD5 + , Express TGF-β and Co-localize with CD4 + Foxp3 + T Cells

doi: 10.1038/mi.2012.42

Figure Lengend Snippet: Panel A: CD4 + CD25 − T cells from spleens of naïve mice were co-cultured for 5 days without or with stimulation by soluble anti-CD3 and anti-CD28. Stimulated cells were also co-cultured with either irradiated CD19 + CD5 + B cells or CD19 + CD5 − B cells from LIT HLNs. T cell Foxp3 expression was increased when cultured with LIT HLN CD5 + B cells versus with LIT CD5 − B cells or anti-CD3/CD28 alone. Data represent mean ± SEM values of 3–4 animals per group; ** indicates p < 0.01 vs. other groups by ANOVA. Panel B: Foxp3 + Treg expression was compared in LIT control and JhD −/− mice. Control mice showed expansion of Foxp3 + Treg cells in hilar nodes (black bars) but not inguinal nodes (striped bars), but this local expansion did not occur in the JhD −/− mice. n = 5 mice per group; * indicates p < 0.005 between CRL and JhD −/− mice. Panel C: Two days before a week of daily OVA aerosol exposures, OVA-sensitized mice received tail vein injections of saline or specific B cell populations. Mice receiving LIT HLN CD5 + B cells demonstrated increased numbers of Foxp3 + T cells in their BAL, as compared to control (saline) mice or mice receiving LIT HLN CD5 − B cells or LIT spleen CD5 + B cells. n = 5–11 mice per group; * indicates p < 0.05 vs. other groups by ANOVA. Panel D: For all groups of mice in panel C, there was a direct correlation between the number of CD4 + Foxp3 + Treg cells and the number of CD19 + CD5 + B cells in BAL (dashed line; r = 0.56; p < 0.005). This relationship also held for the LIT HLN CD5 + recipient animals alone (solid line; r = 0.73; p < 0.05). In contrast, there was no association between CD4 + CD25 + Foxp3 − Teff cells and CD19 + CD5 + B cells in BAL (r = 0.28; p > 0.10; data not shown).

Article Snippet: B cells from HLNs and spleens of LIT mice were positively selected using a mouse CD19 + isolation kit (Miltenyi Biotech, Auburn, CA).

Techniques: Cell Culture, Irradiation, Expressing, Control, Aerosol, Saline

Journal: Mucosal immunology

Article Title: Regulatory B Cells from Hilar Lymph Nodes of Tolerant Mice in a Murine Model of Allergic Airway Disease are CD5 + , Express TGF-β and Co-localize with CD4 + Foxp3 + T Cells

doi: 10.1038/mi.2012.42

Figure Lengend Snippet: CD5 + B cells were isolated from hilar nodes (black bars) and inguinal nodes (gray bars) at different stages (Naïve, Sensitized, AAD and LIT) of the OVA model. Panel A depicts representative flow cytometry dot plots of CXCR4 expression on CD19 + CD5 + B cells. Panel B demonstrates increased CXCR4 + CD5 + B cells in hilar compared to inguinal lymph nodes at all stages of the model, and expansion of hilar node CXCR4 + CD5 + B cells during AAD and LIT. Panel C demonstrates similar CXCR5 expression by CD5 + B cells in both tissues and at all stages of the model. Data represent the mean ± SEM; n = 8–12 in each group (A, B); * indicates p < 0.05 as compared to Naïve and Sensitized groups in the HLN and to all groups in the ILN; ! indicates p < 0.05 as compared to all groups in the ILN; † indicates p < 0.05 between HLN and ILN in Naïve and Sensitized groups; †† indicates p < 0.005 between HLN and ILN in AAD and LIT groups.

Article Snippet: B cells from HLNs and spleens of LIT mice were positively selected using a mouse CD19 + isolation kit (Miltenyi Biotech, Auburn, CA).

Techniques: Isolation, Flow Cytometry, Expressing

Journal: Mucosal immunology

Article Title: Regulatory B Cells from Hilar Lymph Nodes of Tolerant Mice in a Murine Model of Allergic Airway Disease are CD5 + , Express TGF-β and Co-localize with CD4 + Foxp3 + T Cells

doi: 10.1038/mi.2012.42

Figure Lengend Snippet: OVA-sensitized mice received tail vein injections of saline or specific B cell populations (0.2 × 10 6 CD19 + CD5 + or 1.0 × 10 6 CD19 + CD5 − cells) 2 days before a week of daily exposure to 1% OVA aerosols. Mice receiving the LIT HLN CD5 + B cells (solid bars) developed attenuated AAD, with less relative (Panel A) and absolute (Panel B) airway eosinophilia relative to control (saline) mice or mice receiving LIT HLN CD5 − B cells or LIT spleen CD5 + B cells. Data represent mean ± SEM values of 5–11 mice per group; * indicates p < 0.05 vs. other groups by ANOVA.

Article Snippet: B cells from HLNs and spleens of LIT mice were positively selected using a mouse CD19 + isolation kit (Miltenyi Biotech, Auburn, CA).

Techniques: Saline, Control

Journal: Pharmacology & therapeutics

Article Title: Current modalities in cancer immunotherapy: immunomodulatory antibodies, CARs and vaccines

doi: 10.1016/j.pharmthera.2017.03.008

Figure Lengend Snippet: CAR clinical trials by target antigen

Article Snippet: Univ.|Takara Phase1,2 18 NCT02134262 CD19 CD28z T cells Retro B-NHL Kite Pharma Phase1,2 124 NCT02348216 CD19 CD28z T cells Retro B-ALL Juno Phase 2 90 NCT02535364 CD19 CD28z T cells Retro B-NHL Kite Pharma Phase 2 70 NCT02601313 CD19 CD28z T cells Retro B-ALL Kite Pharma Phase1,2 75 NCT02614066 CD19 CD28z T cells Retro B-ALL Kite Pharma Phase1,2 75 NCT02625480 CD19 CD28z T cells Retro B-NHL Xuzhou Med.

Techniques: Clinical Proteomics, Virus