Journal: Cell reports

Article Title: Integration of T helper and BCR signals governs enhanced plasma cell differentiation of memory B cells by regulation of CD45 phosphatase activity

doi: 10.1016/j.celrep.2021.109525

Figure Lengend Snippet: (A) Flow cytometry dot plot showing CD45 phosphatase activity versus CD27 expression on gated CD19 + human peripheral B cells. (B and C) CD45 phosphatase activity (B) and CD45 surface expression (C) of CD27 − (blue) and CD27 + B cells (red). Numbers in histograms represent CD45 activity (pCAP-SP1) (B) or CD45 surface expression (C) as the mean fluorescence intensity (MFI) ratio of CD27 + /CD27 − B cells. Bottom graphs: pCAP-SP1 or CD45 surface expression (MFI) in CD27 + relative to CD27 − B cells. (D) CD45 expression versus CD45 phosphatase activity in gated CD27 + MBCs; CD45 hi and CD45 lo expression gates are shown. (E) CD45 phosphatase activity and (F) CD45 expression in CD27 + MBCs expressing low (blue open histogram) or high (red open histogram) levels of surface CD45 compared to CD27 − B cells (filled blue histogram). Graphs show pCAP-SP1 or CD45 MFI relative to CD27 − B cells. n = 12. Related to . ****p < 0.0001.

Article Snippet: FITC Mouse monoclonal anti human CD27 (clone MT271) , Miltenyi Biotec , Cat# 130–093-184; RRID:AB_1036205.

Techniques: Flow Cytometry, Activity Assay, Expressing, Fluorescence

Journal: Cell reports

Article Title: Integration of T helper and BCR signals governs enhanced plasma cell differentiation of memory B cells by regulation of CD45 phosphatase activity

doi: 10.1016/j.celrep.2021.109525

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: FITC Mouse monoclonal anti human CD27 (clone MT271) , Miltenyi Biotec , Cat# 130–093-184; RRID:AB_1036205.

Techniques: Negative Control, Recombinant, Purification, Staining, Gene Expression, Lysis, Immunoprecipitation, Plasmid Preparation, Enzyme-linked Immunosorbent Assay, Blocking Assay, Cell Isolation, Software, Microscopy

Journal: Cell Reports Methods

Article Title: Development of an HIV reporter virus that identifies latently infected CD4 + T cells

doi: 10.1016/j.crmeth.2022.100238

Figure Lengend Snippet:

Article Snippet: Anti-human CD27 (REA499) 155Gd, coupled in house , Miltenyi Biotec , Cat#: 130122295, RRID: AB_2801876.

Techniques: Virus, Plasmid Preparation, Recombinant, Cell Isolation, Staining, Software

Journal: bioRxiv

Article Title: Disruptors of sestrin-MAPK interactions rejuvenate T cells and expand TCR specificity

doi: 10.1101/2024.05.17.594698

Figure Lengend Snippet: (A) Senescence-associated β-galactosidase expression in DOS-treated (DOS-juvenated) or untreated T sen . Cells were purified and cultured for one week in the presence of anti-CD3 (0.5 μg/mL) and rh-IL-2 (5 ng/mL), then stained to detect β-galactosidase activity. Representative image on the inverted phase-contrast microscope (left) and relative quantification (right, n = 8 donors). (B) Population doublings of human T sen (transduced with irrelevant scramble) and sestrin null CD4 + T sen (transduced by triple lentiviral depletion of sestrins) and cultured as in (A) (left). Donor-matched CD27 + CD28 + CD4 + T cells (herafter, T erl ) were cultured in parallel but activated with anti-CD3 and anti-CD28 ( n = 5 donors). Cells were cultured over two weeks with restimulation every 7 days. DOS-driven population doublings (right) were calculated as delta between DOS treated and DOS untreated T cells, with or without depletion of sestrins. (C) DOS-juvenation of human CD4 + T cells. Terminally differentiated effector memory CD45RA - CD28 - CD27 - CD4 + T cells (hereafter, T EM ) were purified and cultured over 20 days, as in (A). At day 1 (18 hours), 7, and 21 cell phenotypes were assessed by flow cytometry. Quantifications of rejuvenated stem like (CD28 + CD45RA + CCR7 + CD95 + CD62L + TCF1 + ) among human CD4 + T cells are shown (left; n = 5 donors). Decay of T EM and CD28 - CD27 - CD45RA + CD4 + T cells (hereafter, T EMRA ) undergoing rejuvenation is shown (right). (D) Adoptive transfer of DOS-juvenated T cells, experimental design. Donor T cells were derived from twenty-month-old mice 15 days after Fluad vaccination with or without DOS treatment (0.1 mg/Kg throughout), labeled with Cell Trace Violet (CTV) dye or congenic CD45.1 tracking, then transferred into young naïve CD45.2 recipients (3 months). In parallel, young mice (3 months) were used as young donor control. Recipient animals were rested for 28 days, then analysed for donor T cell persistence and maintenance of stem phenotype after transfer. (E) Maintenance of donor DOS-juvenated CD45.1 CD4 + T cells, their aged-matched controls, and that of young donor T cells, 28 days after transfer (day 43) in recipient mouse lymph nodes. Representative flow cytometry plots and poled data ( n = 5 mice per group) are shown. (F) Assessment of mouse T cell memory programs in stem like transferred T cells (among CD45.1 CD44 - CD62L + CD95 + CD4 + T cells) and terminally differentiated cells (TE, among CD45.1 CD44 - CD62L - CD4 + T cells) following adoptive transfer as in (D) ( n = 5 mice). (G) IL7R gene expression and (H) CD95 and Sca-1 mean fluorescent intensity (MFI; throughout) in stem cells derived from CD4 + CD45.1 + transferred stem T cells among lymph nodes of recipient CD45.2 mice, 28 days after transfer ( n = 5 mice). (I) Representative plots of CD45.1 transferred cells in quiescent state before and after transfer assessed by cycle related intra-nuclear Ki67 staining. Representative of n = 5 mice per group. (J) G1 (Ki67 + ) to G0 (Ki67 - ) transition in stem like CD45.1 CD4 + T cells before and after adoptive transfer as indicated ( n = 5 mice per group). (K) Assessment of T cell longevity following DOS-juvenation. Cells from lymph nodes were stained using the Annexin-PI Apoptosis detection Kit 28 days after transfer. Representative FACS plot (left) and quantification of dead CD45.1 + CD4 + transferred T cells (right). (L) DOS-juvenated T cell maintenance, in vivo model. In (A and B, right) two tailed paired T test was used. In (B, left-C, E-H and J-K) one-way Anova with Bonferroni post-correction for multiple comparisons was used, *p<0,05, **P<0,01; ***P<0,001; ****P<0,0001. Error bars indicate SEM.

Article Snippet: Primary human CD4 + T cells were isolated using CD4 MicroBeads (130-045-101; Miltenyi); primary human senescent CD4 + CD27 - CD28 - T cells (thereafter T sen ) were isolated using CD4 + T Cell Isolation Kit (130-096-533; Miltenyi) followed by depletion of CD27 + and CD28 + T cells using CD27 MicroBeads (130-051-601; Miltenyi) and CD28 MicroBead Kit (130-093-247; Miltenyi).

Techniques: Expressing, Purification, Cell Culture, Staining, Activity Assay, Microscopy, Quantitative Proteomics, Transduction, Flow Cytometry, Adoptive Transfer Assay, Derivative Assay, Labeling, Control, Gene Expression, In Vivo, Two Tailed Test

Journal: Cell reports

Article Title: High-dimensional profiling clusters asthma severity by lymphoid and non-lymphoid status

doi: 10.1016/j.celrep.2021.108974

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Anti-Human CD27 (L128)-167Er , Fluidigm , Cat# 3167006B.

Techniques: Purification, Control, Antibody Labeling, Flow Cytometry, Software, Staining, Blocking Assay

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: To block CD70–CD27 binding, cells were incubated in the presence of anti-CD27 blocking antibody (R&D systems, MAB382) or isotype mouse IgG1 (R&D systems, MAB002) for 30 min at 4 °C before binding.

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: To block CD70–CD27 binding, cells were incubated in the presence of anti-CD27 blocking antibody (R&D systems, MAB382) or isotype mouse IgG1 (R&D systems, MAB002) for 30 min at 4 °C before binding.

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

Journal: Cell Reports Medicine

Article Title: Tumor evolution and immune microenvironment dynamics in primary and relapsed mantle cell lymphoma

doi: 10.1016/j.xcrm.2025.102318

Figure Lengend Snippet: High CD70 expression and strong CD70-CD27 interaction in MC3 relapse tumors (A) CD70-CD27 interaction between malignant subclusters and T subsets. (B) CD70 expression levels across MC3 malignant subclusters. Each box represents the interquartile range (IQR) with a line indicating the median. Whiskers extend to the minimum and maximum values within 1.5 times the IQR, excluding outliers. (C) UMAP of sequenced single cells in MC3 samples colored by cell type and CD70 and CD27 expression. (D) Immunohistochemistry images of MC3 primary and relapsed tumors stained with an anti-CD70 antibody. Scale bars, 100 μm (top left micrograph), 2 mm (top right micrograph), 100 μm (bottom zoomed-in micrographs). (E) Multiplex immunofluorescence staining of the MC3Ri sample showing high CD70 expression in malignant cells (row 1: CD20, CCND1, and CD70) and the presence of CD4 + FOXP3 + and CD8 + PD-1 + cells in the tumor microenvironment. Scale bars, 2 mm (overview micrographs) and 50 μm (right zoomed-in micrographs). See also .

Article Snippet: The samples were incubated with primary antibodies specific for human CD70 (MAB2738, clone 301731, 1:100 dilution, R&D Systems) in a DAKO autostainer Link 48 instrument via an Envision Flex detection kit (DAKO) according to the manufacturer’s instructions.

Techniques: Expressing, Immunohistochemistry, Staining, Multiplex Assay, Immunofluorescence

Journal: British Journal of Cancer

Article Title: Phase II study of everolimus with biomarker exploration in patients with advanced gastric cancer refractory to chemotherapy including fluoropyrimidine and platinum

doi: 10.1038/bjc.2012.47

Figure Lengend Snippet: IHC markers (at baseline biopsy) vs progression-free survival (PFS) and overall survival (OS)

Article Snippet: Briefly, 4 μ m thick whole tissue sections were transferred onto poly-L-lysine-coated adhesive slides and dried at 74°C for 30 min. After standard heat epitope retrieval for 1 h in ethylene diamine tetraacetic acid, pH.8.0, in the autostainer, samples were incubated with antibodies against phosphorylated mTOR (pmTOR, 1 : 200 dilution, clone 49F9, rabbit monoclonal, Cell Signaling Technology, Danvers, MA, USA), pS6K1 (1 : 100 dilution, clone E175, rabbit monoclonal, Epitomics, Burlingame, CA, USA), pS6 at Ser 235/6 (pS6 Ser235/6 , 1 : 150 dilution, clone 91B2, rabbit monoclonal, Cell Signaling Technology), pS6 at Ser 240/4 (pS6 Ser240/4 , 1 : 200 dilution, rabbit polyclonal, Cell Signaling Technology) and p4EBP1 (1 : 400 dilution, clone 236B4, rabbit monoclonal, Cell Signaling Technology).

Techniques:

Journal: iScience

Article Title: PD-L1 + CD8 + T cells enrichment in lung cancer exerted regulatory function and tumor-promoting tolerance

doi: 10.1016/j.isci.2022.103785

Figure Lengend Snippet:

Article Snippet: anti-human CD27 -150Nd , Fluidigm , Cat# 3150017B.

Techniques: Recombinant, Antibody Labeling, Enzyme-linked Immunosorbent Assay, Sequencing, Software

Journal: Cell Reports Medicine

Article Title: Response and recurrence correlates in individuals treated with neoadjuvant anti-PD-1 therapy for resectable oral cavity squamous cell carcinoma

doi: 10.1016/j.xcrm.2021.100411

Figure Lengend Snippet:

Article Snippet: CD27 (L128) , Fluidigm , Cat# 3155001B, RRID: AB_2687645.

Techniques: Recombinant, Isolation, Mass Cytometry, Software