Journal: Molecular Systems Biology

Article Title: Molecular profiling reveals features of clinical immunity and immunosuppression in asymptomatic P. falciparum malaria

doi: 10.15252/msb.202110824

Figure Lengend Snippet: PBMCs from P. falciparum symptomatic ( n = 16) and asymptomatic ( n = 24) infected individuals as well as healthy immune controls ( n = 24) were stained with a panel of metal‐labeled antibodies and analyzed by CyTOF. Manual gating was used to select the following populations before FlowSOM clustering: A Individual T H1 ‐like memory CD4 + T cells (CD19 − CD3 + CD4 + CD45RA − CCR6 − CXCR3 + ), T H2 ‐like memory CD4 + T cells (CD19 − CD3 + CD4 + CD45RA − CCR6 − CXCR3 − ), circulating memory T FH cells (CD19 − CD3 + CD4 + CD45RA − CXCR5 + ) memory CD4 + T cells. B Classical (CD3 − CD19 + CD20 + CD10 − CD27 + CD21 + ), atypical (CD3 − CD19 + CD20 + CD10 − CD27 − CD21 − ), and activated MBCs (CD3 − CD19 + CD20 + CD10 − CD27 + CD21 − ).

Article Snippet: 167Er‐conjugated mouse anti‐human CD27 , Fluidigm , Clone L128; Cat# 3167006B.

Techniques: Infection, Staining, Labeling

Journal: Molecular Systems Biology

Article Title: Molecular profiling reveals features of clinical immunity and immunosuppression in asymptomatic P. falciparum malaria

doi: 10.15252/msb.202110824

Figure Lengend Snippet: PBMCs from P. falciparum symptomatic ( n = 16) and asymptomatic ( n = 24) infected individuals as well as healthy immune controls ( n = 24) were stained with a panel of metal‐labelled antibodies and analyzed by CyTOF. t‐distributed Stochastic Neighbor Embedding (tSNE) analysis was performed and FlowSOM clustering was used to identify individual cell subpopulations within gated: T H1 ‐like memory CD4 + T cells (CD19 − CD3 + CD4 + CD45RA − CCR6 − CXCR3 + ) T H2 ‐like memory CD4 + T cells (CD19 − CD3 + CD4 + CD45RA − CCR6 − CXCR3 − ) Circulating memory T FH cells (CD19 − CD3 + CD4 + CD45RA − CXCR5 + ) Classical MBCs (CD3 − CD19 + CD20 + CD10 − CD27 + CD21 + ) Atypical MBCs (CD3 − CD19 + CD20 + CD10 − CD27 − CD21 − ) Activated MBCs (CD3 − CD19 + CD20 + CD10 − CD27 + CD21 − ) The tSNE plots in the top panel display cell density and represent the pooled data for each group, while the lower panel shows a projection of the FlowSOM clusters on a tSNE plot. Heatmaps show the median marker expression for each FlowSOM cluster.

Article Snippet: 167Er‐conjugated mouse anti‐human CD27 , Fluidigm , Clone L128; Cat# 3167006B.

Techniques: Infection, Staining, Marker, Expressing

Journal: Molecular Systems Biology

Article Title: Molecular profiling reveals features of clinical immunity and immunosuppression in asymptomatic P. falciparum malaria

doi: 10.15252/msb.202110824

Figure Lengend Snippet: P. falciparum symptomatic ( n = 30, SM) and asymptomatic ( n = 40, AM) infected individuals, as well as light‐microscopy and PCR parasite‐negative healthy immune controls ( n = 31, HC) were recruited for the study (full bars). Subsets of 5–6 samples per group were selected for PBMC transcriptional profiling (striped bars). A–F Clinical parameters determined in the study included: age (A), gender (B), parasitemia (C), hemoglobin (g/dl blood) (D), hematocrit (E), and platelet count (F). G–M Antibody responses against the following antigens were evaluated in the study which included: P . falciparum parasite lysate (G), EBA‐175 (H), EBA‐140 (I), PfRh2 (J), PfRh4 (K), PfRh5 (L), and PfRipr (M). N–T Percentage of cell populations identified by CyTOF with statistically significant odds ratios are shown: CXCR3 + PD‐1 + T H1 T FH cells (N), IgM + classical MBCs (O), IgM + atypical MBCs (P), CD25 low CD27 low CD4 + T H2 memory cells (Q), IgD + IgM low classical MBCs (R), Isotype switched T‐bet + atypical MBCs (S), and IgD + IgM + atypical MBCs (T). Data information: Boxes represent the 25 th to 75 th percentiles, whiskers show the range (minimum to maximum), and lines represent the median. Significance was determined by the Mann–Whitney test, * P < 0.05, **** P < 0.01, **** P < 0.005, **** P < 0.001.

Article Snippet: 167Er‐conjugated mouse anti‐human CD27 , Fluidigm , Clone L128; Cat# 3167006B.

Techniques: Infection, Light Microscopy, MANN-WHITNEY

Journal: Molecular Systems Biology

Article Title: Molecular profiling reveals features of clinical immunity and immunosuppression in asymptomatic P. falciparum malaria

doi: 10.15252/msb.202110824

Figure Lengend Snippet: A, B Hierarchical clustering heatmap (A) and bar plots showing enriched GO pathways (B) using genes upregulated in symptomatic individuals that were significantly correlated (Benjamini–Hochberg adjusted P < 0.05) with IgM + activated MBCs. C, D Hierarchical clustering heatmap (B) and bar plots showing enriched GO pathways (D) using genes upregulated in symptomatic individuals that were significantly correlated with T‐bet + CD4 + T cells. E Hierarchical clustering heat map of genes upregulated in asymptomatic P . falciparum‐ infected individuals that were significantly correlated (Benjamini–Hochberg adjusted P < 0.05) with CD27 + T H2 CD4 + cells.

Article Snippet: 167Er‐conjugated mouse anti‐human CD27 , Fluidigm , Clone L128; Cat# 3167006B.

Techniques: Infection

Journal: Molecular Systems Biology

Article Title: Molecular profiling reveals features of clinical immunity and immunosuppression in asymptomatic P. falciparum malaria

doi: 10.15252/msb.202110824

Figure Lengend Snippet:

Article Snippet: 167Er‐conjugated mouse anti‐human CD27 , Fluidigm , Clone L128; Cat# 3167006B.

Techniques: Control, Enzyme-linked Immunosorbent Assay, Marker, Infection, Purification, Flow Cytometry, Blocking Assay, Generated, Recombinant, Saline, Staining, Software, Cytometry

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: PLoS ONE

Article Title: Defective Expression and Function of the Leukocyte Associated Ig-like Receptor 1 in B Lymphocytes from Systemic Lupus Erythematosus Patients

doi: 10.1371/journal.pone.0031903

Figure Lengend Snippet: Panel A. Percentage of LAIR1 + (black bar) or LAIR1 − B (grey bar) cells in naïve (CD27 − ) or memory (CD27 + ) cell subsets. Results are determined by triple immunofluorescence assay using specific mAbs to CD20, LAIR1 and CD27 followed by isotype specific GAM conjugated with Alexafluor674 (CD20) or PE (LAIR1) or Alexafluor488 (CD27) on HD or SLE or MCTD or SSc patients. Panel B. Effect of B cell activation on LAIR1 expression. Highly purified healthy B cells were stimulated with sIgM or PWM or MALP2, as indicated, and expression of LAIR1 and CD20 was analyzed as in A. Results are representative of seven independent experiments. Panel C. Expression of LAIR1 on T cells from healthy donors (HD), SLE, MCTD and SSc patients. PBMC were stained with anti-CD3 (JT3A, IgG2a) and anti-LAIR1 (IgG1) mAb followed by anti-isotype specific GAM conjugated with Alexafluor647 or PE respectively. Results are expressed as percentages of CD3 + LAIR1 + (dark grey bar) or CD3 + LAIR1 − (light grey bar) T cells.

Article Snippet: Anti-human CD146 (clone P1H12, IgG1) and unrelated isotype matched control mAbs were from BD Pharmingen, anti-CD90 (clone Thy-1A1, IgG2a) and anti-HMGB1 (clone 115603, IgG2b) mAbs were from R&D System Inc. (Minneapolis, USA) and anti-CD27 mAb (clone LT27, IgG2a) was from Serotec (Oxford, UK).

Techniques: Immunofluorescence, Activation Assay, Expressing, Purification, Staining

Journal: PLoS ONE

Article Title: Defective Expression and Function of the Leukocyte Associated Ig-like Receptor 1 in B Lymphocytes from Systemic Lupus Erythematosus Patients

doi: 10.1371/journal.pone.0031903

Figure Lengend Snippet: A. Immunoglobulin production of the indicated isotype (A or G or M) from healthy donors (HD) or SLE patients was analyzed with specific ELISA kit in cell culture supernatant after stimulation for 5 d of PBMC with pokeweed mitogen alone (PWM) (left) and upon cross-linking of LAIR1 (PWM LAIR1-XL) (right). Results are expressed as ng/ml/10 5 CD20 + B cells. Ig production of PBMC from HD (n = 25) is shown for comparison. In the right panel the statistical significance was shown between HD+PWM and HD+PWM-LAIR1-XL, or HD and SLE2 or SLE1 groups in PWM-LAIR1-XL culture condition. Panel B. PBMC of HD, (n = 8, left) or SLE patients (n = 10 from SLE2 group, right) were incubated for 5 d on collagen coated plates. Then SN were analyzed for the presence of the human IgM, IgG and IgA by ELISA. In some experiments, F(ab′) 2 of anti-LAIR1 mAb (5 µ g/ml) to compete with the interaction of surface LAIR1 and collagen or an unrelated mAb matched for the isotype as control mAb (5 µ g/ml) was added at the onset of cell culture. Results are expressed as ng/ml/10 5 CD20 + B cells as mean±SD. * p<0.001 vs basal production of Ig. **p<0.001 vs Ig production of PBMC on collagen coated plates. In the right panel is indicated the statistical significance of Ig production in the culture condition PBMC+collagen in SLE2 patients vs HD.

Article Snippet: Anti-human CD146 (clone P1H12, IgG1) and unrelated isotype matched control mAbs were from BD Pharmingen, anti-CD90 (clone Thy-1A1, IgG2a) and anti-HMGB1 (clone 115603, IgG2b) mAbs were from R&D System Inc. (Minneapolis, USA) and anti-CD27 mAb (clone LT27, IgG2a) was from Serotec (Oxford, UK).

Techniques: Enzyme-linked Immunosorbent Assay, Cell Culture, Incubation

Journal: PLoS ONE

Article Title: Defective Expression and Function of the Leukocyte Associated Ig-like Receptor 1 in B Lymphocytes from Systemic Lupus Erythematosus Patients

doi: 10.1371/journal.pone.0031903

Figure Lengend Snippet: A, LMSC from a representative reactive lymph node were surface stained with the indicated mAbs (first row) followed by PE-conjugated anti-isotype specific goat anti-mouse antiserum. Control: cells stained with an unrelated mAb followed by GAM as negative control. Second row: LMSC were cytoplasmic stained after fixation and permeabilization with mAbs to the indicated molecules (ALP: alkaline phosphatase, BSP: bone sialoprotein, collagen or vimentin) followed by PE-conjugated GAM. Results are expressed as log red fluorescence intensity vs number of cells. In each panel are indicated the percentages of positive cells above the horizontal bar set on negative control (first subpanel on the left of each row). B. left: bright field (BF) of LMSC from a representative reactive lymph node (upper left), staining with anti-collagen mAb (upper right, red) without cell permeabilization and the respective negative controls (BF neg control, neg control). B right: staining of LMSC with anti-HLA-I (surface, green), anti-prolyl-4-hydroxylase (P4H, cytoplasmic, red) and anti-HMGB1 mAb (nucleus, blue) analyzed by confocal microscopy. Merge analysis is also shown. 400× (left), 600× (right) magnification. White Bars: 10 µ m; reactivity for collagen is disposed in large and concentrated regions (upper left); the white arrows indicate the intracytoplasmic reactivity for P4H (upper right). C. PBMC of healthy donors (HD, n = 7, left) or SLE patients (n = 9 from SLE2 group, right) were incubated for 5 d on collagen-producing mesenchymal stromal cells (MSC) from reactive lymph node coated plates. Then SN were analyzed for the presence of the human IgM, IgG and IgA by ELISA. In some experiments, F(ab′) 2 of anti-LAIR1 mAb (5 µ g/ml) to compete with the interaction of surface LAIR1 and collagen-producing MSC or an unrelated mAb matched for the isotype as control mAb (5 µ g/ml) was added at the onset of cell culture. Results are expressed as ng/ml/10 5 CD20 + B cells as mean±SD. * p<0.001 vs basal production of Ig. ** p<0.001 vs Ig production on LMSC coated plates. In the right panel is indicated the statistical significance of Ig production in the culture condition PBMC+LMSC in SLE2 patients vs HD.

Article Snippet: Anti-human CD146 (clone P1H12, IgG1) and unrelated isotype matched control mAbs were from BD Pharmingen, anti-CD90 (clone Thy-1A1, IgG2a) and anti-HMGB1 (clone 115603, IgG2b) mAbs were from R&D System Inc. (Minneapolis, USA) and anti-CD27 mAb (clone LT27, IgG2a) was from Serotec (Oxford, UK).

Techniques: Staining, Negative Control, Fluorescence, Confocal Microscopy, Incubation, Enzyme-linked Immunosorbent Assay, Cell Culture

Journal: Immunity

Article Title: Natural parasite exposure induces protective human anti-malarial antibodies

doi: 10.1016/j.immuni.2017.11.007

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Cells were stained using CSP-Alex647 (1:20; 0.6mg/ml) or MSP3-Alexa647 (1:20; 0.3mg/ml) and the following antibodies: mouse anti-human CD19-PeCy7 (1:20, eBioscience), mouse anti-human CD27-FITC (1:5, BD) and mouse anti-human IgG-Biotin (1:200, BD).

Techniques: Recombinant, Diagnostic Assay, Affinity Chromatography, Labeling, Plasmid Preparation, Nested PCR, Clone Assay, Sequencing, Real-time Polymerase Chain Reaction, Software

Journal: eLife

Article Title: Cell-density independent increased lymphocyte production and loss rates post-autologous HSCT

doi: 10.7554/eLife.59775

Figure Lengend Snippet:

Article Snippet: Antibody , Anti-human CD27-APC (Mouse IgG1, κ) RRID: AB_469371 , eBioscience , Cat# 17-0279-42 Clone: O343 , ‘(1:25)’.

Techniques: Luminex, Software

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: 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