Journal: Clinical and Experimental Immunology

Article Title: Renal‐infiltrating CD11c + cells are pathogenic in murine lupus nephritis through promoting CD4 + T cell responses

doi: 10.1111/cei.13017

Figure Lengend Snippet: Accumulation of a CD11c+ cell population in the kidney of lupus‐prone mice. (a) Immunohistochemistry (IHC) stains of CD11c+ cells (red) on the kidney sections of 4‐month‐old Murphy Roths large (MRL) and MRL/lpr mice. Representative images are shown. Blue, 4',6‐diamidino‐2‐phenylindole (DAPI). (b) Stepwise gating of CD11c+ cells by flow cytometry as CD11c+CD45+Lin(CD3, CD19 and CD49b)–CD11b+ cells from isolated kidney mononuclear cells from MRL/lpr mice. Representative flow cytometry plots are shown. (c) IHC stains of CD11c+ cells (red) and CD11b+ cells (green) on the kidney sections of 4‐month‐old MRL/lpr mice. Representative images of the medulla region are shown. (d–f) The percentages of renal‐infiltrating CD11c+ cells in Lin– population (top row) as gated in (b) and the relative cell count changes of renal‐infiltrating CD11c+ cells (bottom row) in (d) 6‐week‐ and 15‐week‐old MRL/lpr mice, (e) 35‐week‐old New Zealand white (NZW) mice and NZB/W mice, and (f) 6‐week‐, 19‐week‐ and 37‐week‐old MRL mice. *P < 0·05; **P < 0·01; ***P < 0·001, Student's t‐test for (d,e) and one‐way analysis of variance (anova) for (f). Data are shown as mean ± standard error of the mean (s.e.m.); n = 3 mice in each group. [Colour figure can be viewed at wileyonlinelibrary.com]

Article Snippet: Anti‐mouse antibodies used in this study include eBioscience: CD3‐biotin (clone 145–2C11), CD19‐biotin (clone 1D3), CD11c‐biotin (clone N418), CD11c‐PE, CD11b‐PerCP‐Cy5.5 (clone M1/70), CD45‐FITC (clone, 30‐F11), CD45‐PE, CD80‐PE (clone RMMP‐1), CD103‐APC (clone 2E7) and F4/80‐eFluor 450 (clone BM8); Biolegend: CD16–2 (FcgR IV)‐APC (clone 9E9), CD40‐APC (clone 3.23), CD44‐APC‐Cy7 (clone IM7), CD64 (FcgR I)‐PE (clone X54–5/7.1), CD138‐APC (clone 281–2), CD49b‐biotin (clone DX5), programmed death ligand 1 (PD‐L1)‐APC (clone 10F.9G2), inducible co‐stimulator ligand‐phycoerythrin (ICOSL‐PE) (clone HK5.3), immunoglobulin (Ig)G‐APC (clone poly4053), IgG2a‐PE (clone RMG2a‐62), CCR2‐PE (clone SA203G11), CX 3 CR1‐biotin, CX 3 CR1‐PE (clone SA011F11) and CX 3 CR1‐APC (clone SA011F11); BD Biosciences: B220‐V500 (clone RA3–6B2), CD4‐PE‐Cy7 (clone RM4–5), CD8a‐V450 (clone 53–6.7), CD11b‐APC‐Cy7, CD11c‐PerCP‐Cy5.5, CD11c‐BV510 (clone HL3), CD45‐APC‐Cy7 (clone 30‐F11), CD86‐PE (clone GL1), lymphocyte antigen 6 complex (Ly6C)‐PE‐Cy7 (clone AL‐21), I‐E/I‐A‐BV421 (clone M5/114), I‐E/I‐A‐V500 (clone M5/114), OX40L‐BV421 (clone RM134L) and PD‐L2‐BV510 (clone TY25); Miltenyi Biotec: CD3‐APC (clone 145–2C11), CD115‐PE (clone AFS98), anti‐biotin‐FITC, anti‐biotin‐APC and Ly6G‐biotin (clone 1A8).

Techniques: Immunohistochemistry, Flow Cytometry, Isolation, Cell Counting

Journal: Clinical and Experimental Immunology

Article Title: Renal‐infiltrating CD11c + cells are pathogenic in murine lupus nephritis through promoting CD4 + T cell responses

doi: 10.1111/cei.13017

Figure Lengend Snippet: Phenotype and cytokine/chemokine profile of renal‐infiltrating CD11c+ cells. (a–f) The surface mean fluorescent intensity (MFI) of (a) major histocompatibility complex (MHC)‐II, (b) F4/80, (c) CD103, (d) CD115, (e) Lymphocyte antigen 6 complex (Ly6C) and (f) CCR2 on renal‐infiltrating CD11c+ cells (CD11c+CD11b+, red), CD11b–conventional dendritic cells (cDCs) (defined as CD11c+CD11b–MHC‐II+, blue), monocytes (defined as CD11c–CD11b+Ly6ChighSSC‐Hlow, orange) and neutrophils (defined as CD11c–CD11b+Ly6CmidSSC‐Hhigh, green) from 4‐month‐old Murphy Roths large (MRL)/lpr mice as determined by flow cytometry. Representative flow cytometry histograms are shown. *P < 0·05; **P < 0·01; ***P < 0·001, one‐way analysis of variance (anova). Data are shown as mean ± standard error of the mean (s.e.m.), n = 3 mice in each group. (g) Relative transcript levels of selected cytokines and chemokines as determined by reverse transcription–quantitative polymerase chain reaction (RT–qPCR) in bone marrow monocytes [4',6‐diamidino‐2‐phenylindole (DAPI)–CD11c–CD11b+CD115+Ly6Chigh], bone marrow neutrophils (DAPI–Ly6G+CD11b+), splenic CD8+cDCs (DAPI–CD11b–CD11c+CD8+MHC‐II+) and kidney (KN)‐infiltrating CD11c+ cells (DAPI–CD45+Lin–CD11c+CD11b+) sorted from 4‐month‐old MRL/lpr mice. A heat‐map is shown. Red, higher expression level; green, lower expression level; n = 3 mice in each group. [Colour figure can be viewed at wileyonlinelibrary.com]

Article Snippet: Anti‐mouse antibodies used in this study include eBioscience: CD3‐biotin (clone 145–2C11), CD19‐biotin (clone 1D3), CD11c‐biotin (clone N418), CD11c‐PE, CD11b‐PerCP‐Cy5.5 (clone M1/70), CD45‐FITC (clone, 30‐F11), CD45‐PE, CD80‐PE (clone RMMP‐1), CD103‐APC (clone 2E7) and F4/80‐eFluor 450 (clone BM8); Biolegend: CD16–2 (FcgR IV)‐APC (clone 9E9), CD40‐APC (clone 3.23), CD44‐APC‐Cy7 (clone IM7), CD64 (FcgR I)‐PE (clone X54–5/7.1), CD138‐APC (clone 281–2), CD49b‐biotin (clone DX5), programmed death ligand 1 (PD‐L1)‐APC (clone 10F.9G2), inducible co‐stimulator ligand‐phycoerythrin (ICOSL‐PE) (clone HK5.3), immunoglobulin (Ig)G‐APC (clone poly4053), IgG2a‐PE (clone RMG2a‐62), CCR2‐PE (clone SA203G11), CX 3 CR1‐biotin, CX 3 CR1‐PE (clone SA011F11) and CX 3 CR1‐APC (clone SA011F11); BD Biosciences: B220‐V500 (clone RA3–6B2), CD4‐PE‐Cy7 (clone RM4–5), CD8a‐V450 (clone 53–6.7), CD11b‐APC‐Cy7, CD11c‐PerCP‐Cy5.5, CD11c‐BV510 (clone HL3), CD45‐APC‐Cy7 (clone 30‐F11), CD86‐PE (clone GL1), lymphocyte antigen 6 complex (Ly6C)‐PE‐Cy7 (clone AL‐21), I‐E/I‐A‐BV421 (clone M5/114), I‐E/I‐A‐V500 (clone M5/114), OX40L‐BV421 (clone RM134L) and PD‐L2‐BV510 (clone TY25); Miltenyi Biotec: CD3‐APC (clone 145–2C11), CD115‐PE (clone AFS98), anti‐biotin‐FITC, anti‐biotin‐APC and Ly6G‐biotin (clone 1A8).

Techniques: Flow Cytometry, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Expressing

Journal: Clinical and Experimental Immunology

Article Title: Renal‐infiltrating CD11c + cells are pathogenic in murine lupus nephritis through promoting CD4 + T cell responses

doi: 10.1111/cei.13017

Figure Lengend Snippet: CX3CR1 highly expressed on renal‐infiltrating CD11c+ cells but dispensable for their infiltration into the nephritic kidney. (a) The transcript level of CX3CR1 as determined by reverse transcription–quantitative polymerase chain reaction (RT–qPCR) in bone marrow monocytes [4',6‐diamidino‐2‐phenylindole (DAPI)‐CD11c– CD11b+CD115+lymphocyte antigen 6 complex (Ly6C)high], bone marrow neutrophils (DAPI–Ly6G+CD11b+), splenic CD8+ conventional dendritic cells (cDCs) [DAPI–CD11b–CD11c+CD8+ major histocompatibility complex (MHC)‐II+] and kidney (KN)‐infiltrating CD11c+ cells (DAPI–CD45+Lin–CD11c+CD11b+) sorted from 4‐month‐old Murphy Roths large (MRL)/lpr mice. (b) The surface expression of CX3CR1 on renal‐infiltrating CD11c+ cells (CD11c+CD11b+, red), CD11b–cDCs (CD11c+CD11b–MHC‐II+, blue), monocytes (CD11c–CD11b+Ly6ChighSSC‐Hlow, orange) and neutrophils (CD11c–CD11b+Ly6CmidSSC‐Hhigh, green) from 4‐month‐old MRL/lpr mice as determined by flow cytometry. A representative flow cytometry histogram is shown. (c) The expression of CX3CR1 and green fluorescent protein (GFP) by peripheral blood mononuclear cells from the fifth generation of MRL/lpr‐CX3CR1+/+ and MRL/lpr‐CX3CR1gfp/gfp littermate mice as determined by flow cytometry. Representative flow cytometry plots are shown. (d) The absolute number of renal‐infiltrating CD11c+ cells in 15‐week‐old MRL, and the fifth generation of MRL/lpr‐CX3CR1+/+ and MRL/lpr‐CX3CR1gfp/gfp mice as determined by flow cytometry. (e) The proteinuria (PU) scores of the same three groups of mice. (f) The transcript levels of CXCR4, CCR1, CCR10 and chemR23 in renal‐infiltrating CD11c+ cells (DAPI–CD45+Lin–CD11c+CD11b+), monocytes (DAPI–CD45+Lin–CD11c–CD11b+Ly6ChighSSC‐Hlow) and lymphocytes (DAPI–CD45+Lin+) sorted from the same kidneys of 4‐month‐old MRL/lpr mice as determined by RT–qPCR. RL = relative level. *P < 0·05; **P < 0·01; ***P < 0·001, one‐way analysis of variance (anova). Data are shown as mean ± standard error of the mean (s.e.m.), n ≥ 3 mice in each group. [Colour figure can be viewed at wileyonlinelibrary.com]

Article Snippet: Anti‐mouse antibodies used in this study include eBioscience: CD3‐biotin (clone 145–2C11), CD19‐biotin (clone 1D3), CD11c‐biotin (clone N418), CD11c‐PE, CD11b‐PerCP‐Cy5.5 (clone M1/70), CD45‐FITC (clone, 30‐F11), CD45‐PE, CD80‐PE (clone RMMP‐1), CD103‐APC (clone 2E7) and F4/80‐eFluor 450 (clone BM8); Biolegend: CD16–2 (FcgR IV)‐APC (clone 9E9), CD40‐APC (clone 3.23), CD44‐APC‐Cy7 (clone IM7), CD64 (FcgR I)‐PE (clone X54–5/7.1), CD138‐APC (clone 281–2), CD49b‐biotin (clone DX5), programmed death ligand 1 (PD‐L1)‐APC (clone 10F.9G2), inducible co‐stimulator ligand‐phycoerythrin (ICOSL‐PE) (clone HK5.3), immunoglobulin (Ig)G‐APC (clone poly4053), IgG2a‐PE (clone RMG2a‐62), CCR2‐PE (clone SA203G11), CX 3 CR1‐biotin, CX 3 CR1‐PE (clone SA011F11) and CX 3 CR1‐APC (clone SA011F11); BD Biosciences: B220‐V500 (clone RA3–6B2), CD4‐PE‐Cy7 (clone RM4–5), CD8a‐V450 (clone 53–6.7), CD11b‐APC‐Cy7, CD11c‐PerCP‐Cy5.5, CD11c‐BV510 (clone HL3), CD45‐APC‐Cy7 (clone 30‐F11), CD86‐PE (clone GL1), lymphocyte antigen 6 complex (Ly6C)‐PE‐Cy7 (clone AL‐21), I‐E/I‐A‐BV421 (clone M5/114), I‐E/I‐A‐V500 (clone M5/114), OX40L‐BV421 (clone RM134L) and PD‐L2‐BV510 (clone TY25); Miltenyi Biotec: CD3‐APC (clone 145–2C11), CD115‐PE (clone AFS98), anti‐biotin‐FITC, anti‐biotin‐APC and Ly6G‐biotin (clone 1A8).

Techniques: Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Expressing, Flow Cytometry

Journal: Clinical and Experimental Immunology

Article Title: Renal‐infiltrating CD11c + cells are pathogenic in murine lupus nephritis through promoting CD4 + T cell responses

doi: 10.1111/cei.13017

Figure Lengend Snippet: The interaction between renal‐infiltrating CD11c+ cells and renal‐infiltrating CD4+ T cells. (a) The percentages of renal‐infiltrating CD11c+CD11b+ cells, CD11b– conventional dendritic cells (cDCs), monocytes and neutrophils in Lin– population of 6‐ and 15‐week‐old Murphy Roths large (MRL)/lpr mice. CD11c+CD11b+ cells were the predominant population in the kidney of 15‐week‐old mice with active lupus nephritis (LN). (b) Total renal infiltration areas of CD11c+ cells (red) and CD3+ T cells (green) from 3‐, 5‐, 7‐, 9‐, 11‐ and 14‐week‐old MRL/lpr mice as determined by immunohistochemistry (IHC) and ImageJ quantification. RL = relative level. A representative image of the kidney (KN) of a 14‐week‐old MRL/lpr mouse is shown. Bar equals 200 μm. Blue, 4',6‐diamidino‐2‐phenylindole (DAPI). (c,d) The surface level of Fc gamma receptor (FcgR) I, FcgR IV, immunoglobulin (Ig)G, IgG2a, major histocompatibility complex (MHC)‐II, CD86, programmed death‐ligand 1(PD‐L1) and PD‐L2 on renal‐infiltrating CD11c+ cells (red), monocytes (orange) and neutrophils (blue) in the kidney of 4‐month‐old MRL/lpr mice as determined by flow cytometry. Representative flow cytometry histograms are shown. (e) The percentages of renal‐infiltrating CD4+, CD8+ and CD4–CD8–B220+ double‐negative (DN) T cells in total renal‐infiltrating CD3+ T cells of 4‐month‐old MRL/lpr mice as determined by flow cytometry. The gating strategy is shown. (f) Interferon (IFN)‐γ and interleukin (IL)‐17a levels in the culture supernatant of CD4+, CD8+ and DN T cells stimulated with anti‐CD3/CD28 as determined by enzyme‐linked immunosorbent assay (ELISA). (g–i) The percentages of live cells (DAPI–) and proliferating cells [carboxyfluorescein succinimidyl ester (CFSE)low] in renal‐infiltrating CD4+ T cells cultured alone or co‐cultured with renal‐infiltrating CD11c+ cells that were stimulated with anti‐CD3/CD28 and macrophage colony‐stimulating factor (M‐CSF_ in the presence of (h) Toll‐like receptor (TLR)‐7 agonist, Imiquimod or (i) TLR‐9 agonist, oligodeoxynucleotide (ODN) 1585 cytosine–phosphate–guanine (CpG). Representative flow cytometry plots and the gating strategy are shown in (g). (j) IFN‐γ levels in the culture supernatant of renal‐infiltrating CD4+ T cells cultured alone or co‐cultured with renal‐infiltrating CD11c+ cells that are stimulated with anti‐CD3/CD28, M‐CSF and ODN 1585 CpG as determined by enzyme‐linked immunosorbent assay (ELISA). *P < 0·05, **P < 0·01, ***P < 0·001, one‐way analysis of variance (anova) for (a–f) and Student's t‐test for (h–j). Data are shown as mean ± standard error of the mean (s.e.m.), n = 3 mice in each group. [Colour figure can be viewed at wileyonlinelibrary.com]

Article Snippet: Anti‐mouse antibodies used in this study include eBioscience: CD3‐biotin (clone 145–2C11), CD19‐biotin (clone 1D3), CD11c‐biotin (clone N418), CD11c‐PE, CD11b‐PerCP‐Cy5.5 (clone M1/70), CD45‐FITC (clone, 30‐F11), CD45‐PE, CD80‐PE (clone RMMP‐1), CD103‐APC (clone 2E7) and F4/80‐eFluor 450 (clone BM8); Biolegend: CD16–2 (FcgR IV)‐APC (clone 9E9), CD40‐APC (clone 3.23), CD44‐APC‐Cy7 (clone IM7), CD64 (FcgR I)‐PE (clone X54–5/7.1), CD138‐APC (clone 281–2), CD49b‐biotin (clone DX5), programmed death ligand 1 (PD‐L1)‐APC (clone 10F.9G2), inducible co‐stimulator ligand‐phycoerythrin (ICOSL‐PE) (clone HK5.3), immunoglobulin (Ig)G‐APC (clone poly4053), IgG2a‐PE (clone RMG2a‐62), CCR2‐PE (clone SA203G11), CX 3 CR1‐biotin, CX 3 CR1‐PE (clone SA011F11) and CX 3 CR1‐APC (clone SA011F11); BD Biosciences: B220‐V500 (clone RA3–6B2), CD4‐PE‐Cy7 (clone RM4–5), CD8a‐V450 (clone 53–6.7), CD11b‐APC‐Cy7, CD11c‐PerCP‐Cy5.5, CD11c‐BV510 (clone HL3), CD45‐APC‐Cy7 (clone 30‐F11), CD86‐PE (clone GL1), lymphocyte antigen 6 complex (Ly6C)‐PE‐Cy7 (clone AL‐21), I‐E/I‐A‐BV421 (clone M5/114), I‐E/I‐A‐V500 (clone M5/114), OX40L‐BV421 (clone RM134L) and PD‐L2‐BV510 (clone TY25); Miltenyi Biotec: CD3‐APC (clone 145–2C11), CD115‐PE (clone AFS98), anti‐biotin‐FITC, anti‐biotin‐APC and Ly6G‐biotin (clone 1A8).

Techniques: Immunohistochemistry, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Cell Culture

Journal: eLife

Article Title: Short-range interactions between fibrocytes and CD8 + T cells in COPD bronchial inflammatory response

doi: 10.7554/eLife.85875

Figure Lengend Snippet:

Article Snippet: Antibody , Anti-IL-17-PE-Cy7 (mouse monoclonal) , Miltenyi Biotec , Cat. #:130-120-413, RRID: AB_2752086 , FC (1:20).

Techniques: Blocking Assay, Recombinant

Journal: PLoS Pathogens

Article Title: Characteristics of Memory B Cells Elicited by a Highly Efficacious HPV Vaccine in Subjects with No Pre-existing Immunity

doi: 10.1371/journal.ppat.1004461

Figure Lengend Snippet: PBMC samples collected at month 7 (n = 12) were enriched for B cells, divided into two parts, and stained with a multicolor flow cytometry panel to identify classic Bmem and either AF488-BPV or AF488-HPV 16. ( A ) Representative dot plots are shown of the total naive B cell (IgD + CD27 − ) and Bmem (IgD − CD27 + ) frequencies in samples collected from subjects EK1006, EK1027, EK1073 and EK1078 (left column), as well as the frequencies of AF488 + Bmem observed by Ag-specific labeling (middle column) or by negative control labeling (right column). Flow cytometry data were first gated for cell size (forward versus side scatter), to exclude doublets and dead cells, and to include B cells (CD3 − CD19 + ) (not shown). ( B ) AF488-HPV16 + Bmem were observed at a significantly higher frequency than AF488-BPV + Bmem for all subjects' analyzed (**, p<0.005; paired, two-tailed student's t-test).

Article Snippet: Each of the two samples were then washed and re-suspended in 2% FBS-PBS solution and stained with either AF488-HPV 16 psV or AF488-BPV psV, as well as the following fluorescent mAbs for 30 minutes: anti-CD38 APC and anti-IgD PE (Milteyni Biotec), anti-CD3 V500, anti-CD19 APC-Cy7, anti-CD27 PE-Cy7, and anti-CD20 PerCP-Cy5.5 (BD Biosciences, San Jose, California).

Techniques: Staining, Flow Cytometry, Labeling, Negative Control, Two Tailed Test

Journal: iScience

Article Title: In vitro vascularized immunocompetent patient-derived model to test cancer therapies

doi: 10.1016/j.isci.2023.108094

Figure Lengend Snippet:

Article Snippet: Anti-CD20 PE Cy7 , Miltenyi , Cat# 130-111-340; RRID: AB_2656074.

Techniques: Plasmid Preparation, Recombinant, Lysis, RNA Extraction, Amplification, DC Protein Assay, Enzyme-linked Immunosorbent Assay, Sequencing, Software, Fluorescence, Microscopy

Journal: PLoS ONE

Article Title: High Throughput Sequencing Analysis of the Immunoglobulin Heavy Chain Gene from Flow-Sorted B Cell Sub-Populations Define the Dynamics of Follicular Lymphoma Clonal Evolution

doi: 10.1371/journal.pone.0134833

Figure Lengend Snippet: (A) Three patients were included in the analysis: pt1 and pt2 displaying a pattern of direct evolution and pt3 of evolution through a CPC based on HH analysis. Biopsies were collected at different time points: at relapse/transformation (pt1 and pt2) and after a watch wait period (pt3). In pt 1 two additional biopsies (AD1548 and 1787) analyzed only on the SNP array are shown. In pt1 all samples had an identical MC whilst in pt2 the MC sequences from samples R1655 FL1 and R3878 FL2 showed 2 different dominant clones carrying 3 discriminating mutations . Because these 3 bases also differ from the germline sequence a pattern of direct evolution could not be excluded. (B) Twenty-two samples, from 4 flow-sorted sub-populations, included in the deep sequencing study: pre-germinal center (PGC) (IgD+CD38+) white circles, the germinal center centroblasts (CB) (IgD+CD38+CD10+CXCR4+) and centrocytes, (CC) (IgD+CD38+CD10+CXCR4-) light grey circles and memory enriched (ME) (IgD-CD38-) dark grey circles. The 2 CC CD77- sorted sub-population from R0012 t-FL and R1381 FL1 biopsies, which were part of the deep sequencing study but not included in the final analysis are not shown. The FL sequences are depicted as white circles, the t-FL clones as grey circles, the germline sequence as black circles and the putative CPC sequence as a dashed grey circle. The 2 samples not included in the deep sequencing of IgH-VH gene are depicted as dotted circles. Each vertical arrow represents a line of therapy; the left hand vertical bar shows the time of diagnosis and the right hand vertical bar the time of death. FL = Follicular Lymphoma; t-FL = transformed Follicular Lymphoma; WW = watch and wait.

Article Snippet: Four different sub-populations were flow-sorted from reactive tonsils and FL biopsies cryopreserved cell suspensions and fluorescently labeled using the monoclonal antibodies: CD38-PE-Cy7 (clone HB7), CD10-PE (clone HI10a), CXCR4 PE-Cy5 (clone 12G5), IgD biotin (clone IA6-2), (Becton Dickinson; San Jose, CA, USA), CD77 FITC (clone 5B5) and anti-biotin APC (clone Bio3-18E7) (Miltenyi Biotec).

Techniques: Transformation Assay, Clone Assay, Sequencing, Biomarker Discovery

Journal: PLoS ONE

Article Title: High Throughput Sequencing Analysis of the Immunoglobulin Heavy Chain Gene from Flow-Sorted B Cell Sub-Populations Define the Dynamics of Follicular Lymphoma Clonal Evolution

doi: 10.1371/journal.pone.0134833

Figure Lengend Snippet: Lymph nodes cell suspensions from sequential biopsies obtained from patients with FL/t-FL were stained and sorted in 4 different populations according to the expression of IgD, CD38, CD10 and CXCR4. ME: IgD-CD38-; PGC: IgD+CD38+, CB: IgD-CD38+CD10+CXCR4+; CC: IgD-CD38+CD10+CXCR4-. Flow-cytometric identification of the 4 sub-populations in the sample R8403, patient 3 (top), sample R0012, patient 1 (middle) and tonsils (bottom). In the 2 patients samples the numbers indicate the percentage of the gated and sorted subsets whilst in the tonsil example the numbers are representative of 8 independent experiments on as many different biopsies.

Article Snippet: Four different sub-populations were flow-sorted from reactive tonsils and FL biopsies cryopreserved cell suspensions and fluorescently labeled using the monoclonal antibodies: CD38-PE-Cy7 (clone HB7), CD10-PE (clone HI10a), CXCR4 PE-Cy5 (clone 12G5), IgD biotin (clone IA6-2), (Becton Dickinson; San Jose, CA, USA), CD77 FITC (clone 5B5) and anti-biotin APC (clone Bio3-18E7) (Miltenyi Biotec).

Techniques: Staining, Expressing