pbmcs Search Results


  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    Lonza peripheral blood mononuclear cells pbmcs
    ShK-186 inhibits antineutrophil cytoplasmic antibodies targeting proteinase 3 (PR3-ANCA) production in vitro . (A) IgG production after peripheral blood mononuclear cell <t>(PBMC)</t> stimulation with CpG, B cell-activating factor (BAFF), and interleukin (IL)-21 from 5 healthy controls (HCs) (open circles) and 13 granulomatosis with polyangiitis <t>(GPA)</t> patients (gray circles) in the presence and absence of 1 nM ShK-186. (B) PR3-ANCA production after PBMC stimulation with CpG, BAFF, and IL-21 from 13 GPA patients in the presence and absence of 1 nM ShK-186 (* p
    Peripheral Blood Mononuclear Cells Pbmcs, supplied by Lonza, used in various techniques. Bioz Stars score: 99/100, based on 126 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/peripheral blood mononuclear cells pbmcs/product/Lonza
    Average 99 stars, based on 126 article reviews
    Price from $9.99 to $1999.99
    peripheral blood mononuclear cells pbmcs - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    Millipore human peripheral blood mononuclear cells pbmc
    Detection of ApoE isoforms in differentiating hNP1 cells. Differentiating hNP1 cells were exposed to medium alone (Unt), mock-infected <t>PBMC</t> supernatants (Mock), and <t>HIV-infected</t> PBMC supernatants (HIV), and to rApoE3 or rApoE4 at 3 μg/ml, as described in Materials and Methods. At day 16 of differentiation, and 24 h after replenishment of rApoE in the cultures, cells were lysed for quantitative immunoblotting to detect ApoE. Lower blot depicts representative endogenous ApoE detected in cell culture lysates with molecular weight of 34 kDa, and rApoE with molecular weight of 36 kDa. Upper blot corresponds to beta-actin used as an internal control. For comparison, recombinant ApoE3 and ApoE4 were loaded directly on the gel at a 3 ng per lane, as shown. Bar graphs present ApoE signal normalized to beta-actin; each bar represents the mean of two experiments
    Human Peripheral Blood Mononuclear Cells Pbmc, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 145 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human peripheral blood mononuclear cells pbmc/product/Millipore
    Average 99 stars, based on 145 article reviews
    Price from $9.99 to $1999.99
    human peripheral blood mononuclear cells pbmc - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    94
    Axis-Shield Diagnostics peripheral blood mononuclear cells pbmcs
    Comparison of HBV-specific cell responses in the presence or absence of HBeAg. (A) C protein-specific T-cell responses elicited in ELISPOT assay; (B) Cytokine <t>IFN-Y</t> responses in cultured supernatants of <t>PBMCs</t> stimulated by C protein. HBeAg(−): HBeAg-negative subjects; HBeAg(+): HBeAg-positive subjects; HIV+: HIV co-infected; HIV-: not co-infected with HIV.
    Peripheral Blood Mononuclear Cells Pbmcs, supplied by Axis-Shield Diagnostics, used in various techniques. Bioz Stars score: 94/100, based on 1342 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/peripheral blood mononuclear cells pbmcs/product/Axis-Shield Diagnostics
    Average 94 stars, based on 1342 article reviews
    Price from $9.99 to $1999.99
    peripheral blood mononuclear cells pbmcs - by Bioz Stars, 2020-09
    94/100 stars
      Buy from Supplier

    93
    Thermo Fisher pbmc
    The distribution of fragments of mRNA, rRNA, tRNA, miRNA, and others in the small <t>RNA</t> libraries. a The <t>PBMC</t> of heat-stressed cattle. b The PBMC of non-heat-stressed cattle
    Pbmc, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 14117 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pbmc/product/Thermo Fisher
    Average 93 stars, based on 14117 article reviews
    Price from $9.99 to $1999.99
    pbmc - by Bioz Stars, 2020-09
    93/100 stars
      Buy from Supplier

    94
    Becton Dickinson peripheral blood mononuclear cells pbmcs
    Systemic immune response to <t>LCMV-GP33</t> peptide C57BL/6 mice (5 mice/group) were challenged subcutaneously with TC-1 tumor cells and subsequently treated with various combinations of cisplatin, CpG and GP33 peptide as shown. The <t>PBMCs</t> were analyzed 1 week after the last antigen delivery. The presence of GP33-specific CD8+ T cells in PBMCs was analyzed using intracellular cytokine staining for IFN-γ and CD8+ staining followed by flow cytometry analysis. A. Representative flow cytometry contour plot depicting the frequency of IFN-γ-secreting CD8+ T cells among PBMCs after being pulsed with GP33 peptide. B. Bar graph depicting the number of IFN-γ+ CD8+ T cells among PBMCs (mean ±S.E).
    Peripheral Blood Mononuclear Cells Pbmcs, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 94/100, based on 950 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/peripheral blood mononuclear cells pbmcs/product/Becton Dickinson
    Average 94 stars, based on 950 article reviews
    Price from $9.99 to $1999.99
    peripheral blood mononuclear cells pbmcs - by Bioz Stars, 2020-09
    94/100 stars
      Buy from Supplier

    92
    Nycomed peripheral blood mononuclear cells pbmc
    Dose–response curves for <t>ADCC.</t> The procedures for OxLDL incubated and Ab treatment were the same as described in the legend for Fig. . As effectors normal rat <t>PBMC</t> were added to the culture instead of complement. Note increased 51Cr release
    Peripheral Blood Mononuclear Cells Pbmc, supplied by Nycomed, used in various techniques. Bioz Stars score: 92/100, based on 502 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/peripheral blood mononuclear cells pbmc/product/Nycomed
    Average 92 stars, based on 502 article reviews
    Price from $9.99 to $1999.99
    peripheral blood mononuclear cells pbmc - by Bioz Stars, 2020-09
    92/100 stars
      Buy from Supplier

    93
    Biochrom peripheral blood mononuclear cells pbmc
    miR-20b expression in T cells and in <t>PBMC</t> from natalizumab-associated progressive multifocal leukoencephalopathy (PML). (A) PBMC were isolated using a <t>Ficoll</t> gradient and CD4+ T cells were sorted with magnetic beads ( n = 9; randomly selected patients of our initial longitudinal cohort consisting of 17 patients). miR-20b expression (qPCR) was measured in the CD4+ and CD4-negative fraction and compared using t- test. (B) CD4+ cells were activated by plate-bound anti-CD3-antibody and soluble anti-CD28-antibody without further cytokines, termed “Th0”, and with IL-1 (10 ng/mL), IL-6 (10 ng/mL), IL-23 (10 ng/mL) and TGF- β (5 ng/mL), termed “Th17”, as well as freshly isolated CD4+ T cells, termed “fresh”. miR-20b levels were measured by qPCR. (C) miR-20b levels in PBMC from natalizumab-treated patients ( n = 9, left bar) were compared to miR-20b levels in two PBMC samples from patients with natalizumab-associated PML (each patient one bar).
    Peripheral Blood Mononuclear Cells Pbmc, supplied by Biochrom, used in various techniques. Bioz Stars score: 93/100, based on 447 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/peripheral blood mononuclear cells pbmc/product/Biochrom
    Average 93 stars, based on 447 article reviews
    Price from $9.99 to $1999.99
    peripheral blood mononuclear cells pbmc - by Bioz Stars, 2020-09
    93/100 stars
      Buy from Supplier

    89
    Ficoll-Paque Pharmacia peripheral blood mononuclear cell pbmc
    Semi-quantitative <t>PCR</t> analysis of BLV pol detects differences in proviral load between BLV SGV and BLV-rabbits. <t>PBMC</t> were harvested at month indicated after challenge with 1 × 10 8 FLK/BLV cells from indicated rabbits that had been infected with BLV SGV for 24 months or from age-matched naïve controls. DNA from 50,000 PMBC was subjected to nested PCR and gel electrophoresis and stained with ethidium bromide. The position of the 591-bp amplicon is designated by the labeled arrow. In parallel, DNA from 20,000 PBMC was subjected to PCR with primers complementary to β- actin . The position of the 594-bp amplicon is designated by the labeled arrow and the signals were quantified by densitometry.
    Peripheral Blood Mononuclear Cell Pbmc, supplied by Ficoll-Paque Pharmacia, used in various techniques. Bioz Stars score: 89/100, based on 83 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/peripheral blood mononuclear cell pbmc/product/Ficoll-Paque Pharmacia
    Average 89 stars, based on 83 article reviews
    Price from $9.99 to $1999.99
    peripheral blood mononuclear cell pbmc - by Bioz Stars, 2020-09
    89/100 stars
      Buy from Supplier

    99
    Millipore peripheral blood mononuclear cells
    Generation, isolation, and characterization of epidermal growth factor receptor (EGFR)‐specific chimeric antigen receptor (CAR)‐engineered natural killer (NK) <t>cells</t> (EGFR‐CAR NK cells). (A) Flow cytometric analysis of phenotypic and subset composition of <t>peripheral</t> <t>blood</t> <t>mononuclear</t> cells (PBMCs) labeled with anti‐CD3‐PE‐Cy7, anti‐CD56‐PE. (B‐C) Flow cytometric analysis of phenotypic and subset composition of NK cells labeled with anti‐CD3‐PE‐Cy7, anti‐CD56‐PE, and anti‐CD69‐APC‐Cy7. (D) The percentage of CD3‐/CD56 + cells in day 0 PBMCs and day14 PBMC culture. (E) Real‐time PCR and (F) Western blotting analyses of the expression of exogenous CD3ζ in the non‐transduced NK cells, Con‐CAR NK cells, EGFR‐CAR‐1 NK cells, and EGFR‐CAR‐2 NK cells. β‐actin was used as an endogenous control. (G) The transduced NK cells stained with IgG‐FITC and EGFR‐FITC antibodies were detected by flow cytometry
    Peripheral Blood Mononuclear Cells, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 187 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/peripheral blood mononuclear cells/product/Millipore
    Average 99 stars, based on 187 article reviews
    Price from $9.99 to $1999.99
    peripheral blood mononuclear cells - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    93
    Becton Dickinson human pbmcs
    <t>EAT-2</t> over-expression enhances human DC maturation. Human <t>PBMCs</t> were isolated from fresh buffy coat material and were either mock infected or infected with MOI of 10000 with rAd5-EAT2 or rAd5-Null. At 72 hpi, cells were stained for surface expression of CD86 (A), HLA-DR (B), CD80 (C) and CD83 (D) and FACS analysis was performed. Data are representative of at least three independent experiments from three different blood donors with similar results. Samples were plated in quadruplicates. Bars represent mean ± SD. Statistical analysis was completed using a one way ANOVA with a Student–Newman–Keuls post-hoc test. P
    Human Pbmcs, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 93/100, based on 603 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human pbmcs/product/Becton Dickinson
    Average 93 stars, based on 603 article reviews
    Price from $9.99 to $1999.99
    human pbmcs - by Bioz Stars, 2020-09
    93/100 stars
      Buy from Supplier

    92
    Becton Dickinson blood mononuclear cells pbmc
    Intracellular and plasma pharmacokinetic profiles of <t>imatinib</t> in five individual patients (solid lines, plasma concentrations; broken lines, peripheral blood mononuclear cell concentrations)
    Blood Mononuclear Cells Pbmc, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 92/100, based on 177 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/blood mononuclear cells pbmc/product/Becton Dickinson
    Average 92 stars, based on 177 article reviews
    Price from $9.99 to $1999.99
    blood mononuclear cells pbmc - by Bioz Stars, 2020-09
    92/100 stars
      Buy from Supplier

    92
    Valiant peripheral blood mononuclear cells pbmcs
    Isolation and molecular characterization of T cell subsets from <t>PNH</t> patients ( A ) Gating strategy for T cell subsets for cell sorting. <t>PBMCs</t> were stained with ViViD, anti-CD14-Pacific Blue, anti-CD19-Pacific Blue, anti-CD3-BV605, anti-CD4-V500, anti-CD8-APC-H7, anti-CD45RA-PE-Cy7, and anti-CD45RO-APC. Lymphocytes were gated based on their scatter characteristics or forward scatter height vs. forward scatter area. Live T cells were gated based on positive for CD3 and negative for ViViD, CD14, and CD19 to remove dead cells, monocytes, and B cells. CD4 + and CD8 + T cells were then gated based on the characteristic expression patterns of CD45RA and CD45RO. Four different T cell subsets [CD4 + naïve (CD45RA + CD45RO - ), CD4 + memory (CD45RA - CD45RO + ), CD8 + naïve (CD45RA + CD45RO - ), and CD8 + memory (CD45RA - CD45RO + ) T cells] were sorted for RNA-seq. ( B ) Transcriptional validation of the naïve and memory T cell identity by analyzing expression levels of CD4, CD8, CCR5, and EOMES. FPKM (fragments per kilobase of exon per million fragments mapped). * p
    Peripheral Blood Mononuclear Cells Pbmcs, supplied by Valiant, used in various techniques. Bioz Stars score: 92/100, based on 193 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/peripheral blood mononuclear cells pbmcs/product/Valiant
    Average 92 stars, based on 193 article reviews
    Price from $9.99 to $1999.99
    peripheral blood mononuclear cells pbmcs - by Bioz Stars, 2020-09
    92/100 stars
      Buy from Supplier

    99
    GE Healthcare pbmcs
    The systemic cellular immune response to sand fly saliva in volunteers in a cutaneous leishmaniasis-endemic area of Mali. Peripheral blood mononuclear cells <t>(PBMCs)</t> were obtained from 255 individuals aged 11–65 years from the villages of Kemena and Sougoula. IFN-γ, IL-12, IL-10, IL-13, and IL-5 levels were measured in the supernatants of PBMCs from exposed volunteers 96 hours after stimulation with Phlebotomus duboscqi salivary gland homogenate. ( a ) Overall <t>cytokine</t> production by tested individuals. ( b ) Pairwise correlation of expressed cytokines (log-transformed cytokine levels). Numbers in insets indicate the Spearman correlation coefficient and the 95% confidence intervals for each correlation. ( c ) Cytokine responses of individuals in ( a ) stratified by T H 1-predominant, T H 2-predominant, or T H 1/T H 2 mixed responses to sand fly salivary proteins.
    Pbmcs, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 99/100, based on 10970 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pbmcs/product/GE Healthcare
    Average 99 stars, based on 10970 article reviews
    Price from $9.99 to $1999.99
    pbmcs - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    Millipore peripheral blood mononuclear cells pbmcs
    Effects of PGRN on the differentiation of human OCs and PGRN as a predictor of osteoporosis. A , <t>HBMCs</t> and <t>PBMCs</t> were differentiated with RANKL in the absence or presence of PGRN. TRAP + MNCs were counted as described above. B , four sh-RNAs targeting human
    Peripheral Blood Mononuclear Cells Pbmcs, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 3440 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/peripheral blood mononuclear cells pbmcs/product/Millipore
    Average 99 stars, based on 3440 article reviews
    Price from $9.99 to $1999.99
    peripheral blood mononuclear cells pbmcs - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    93
    Corning Life Sciences pbmcs
    Nanoparticle formulation decreases curcumin cytotoxicity. <t>SUPT1</t> cells (Panel A) or stimulated <t>PBMCs</t> (Panel B) were exposed to increasing concentrations (1, 5, 10, 25, 50 and 100 µM) of sol-curcumin, nano-curcumin, azidothymidine (AZT) or nano-apotransferrin (10, 50 and 100 µg) for 16 h, after which cell viability was determined by MTT assay. PBMCs were cultured in the presence of IL-2 (20 IU/ml). Cell viability in the absence of drug was defined as 0% cytotoxicity. Error bars indicate SD. ** P ≤0.01, and ***, P ≤0.001 compared to nano-curcumin. * indicates µg apotransferrin protein that carry equivalent molar concentration of the drug.
    Pbmcs, supplied by Corning Life Sciences, used in various techniques. Bioz Stars score: 93/100, based on 1423 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pbmcs/product/Corning Life Sciences
    Average 93 stars, based on 1423 article reviews
    Price from $9.99 to $1999.99
    pbmcs - by Bioz Stars, 2020-09
    93/100 stars
      Buy from Supplier

    92
    AllCells LLC pbmcs
    Inference and effect of barcode multiplets in single-cell ATAC-seq data. (a) Default t-SNE depiction of public <t>scATAC-seq</t> <t>PBMC</t> 5k dataset. Colors represent cluster annotations from the automated CellRanger output. (b) Quantification of barcodes affected by barcode multiplets for the same dataset (identified by bap). (c) Depiction of two multiplets each composed of 9 oligonucleotide barcodes. Barcodes in each multiplet share a long common subsequence, denoted in black. (d) Visualization of two barcode multiplets from (c) in t-SNE coordinates. (e) Visualization of all implicated barcode multiplets from this dataset. The zoomed panel shows a small group of cells affected by five multiplets, indicated by color. (f) Empirical distribution of the mean restricted longest common subsequence (rLCS) per multiplet. A cutoff of 6 was used to determine either of the two classes of barcode multiplets. (g) Percent difference of the mean log2 fragments between pairs of barcodes within a multiplet. The reported p-value is from a two-sided Kolmogorov–Smirnov test. Boxplots: center line, median; box limits, first and third quartiles; whiskers, 1.5x interquartile range. (h) Overall rates of barcode multiplets from additional scATAC-seq data comparing v1.0 and v1.1 (NextGEM) chip designs.
    Pbmcs, supplied by AllCells LLC, used in various techniques. Bioz Stars score: 92/100, based on 153 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pbmcs/product/AllCells LLC
    Average 92 stars, based on 153 article reviews
    Price from $9.99 to $1999.99
    pbmcs - by Bioz Stars, 2020-09
    92/100 stars
      Buy from Supplier

    93
    Becton Dickinson pbmc isolation
    ( a ) p24 viral antigen levels 10 days post-infection in the supernatant of <t>HIV-1</t> infected PBMCs treated with TIZ (10 μg/mL) or RM-4848 (10 μg/mL). Results obtained with <t>PBMC</t> of 20 different healthy donors are shown. Median values and statistical significance is presented. ( b ) Inhibition (%) of viral replication in PBMC of 20 different healthy donors infected in vitro with HIV-1 in the presence of TIZ- (10 μg/mL) or RM-4848- (10 μg/mL). Median values and statistical significance is presented. ( c ) p24 viral antigen levels in PBMCs infected with HIV-1 in the presence of RM-4848 throughout infection (RM-4848)(10 μg/mL) or in medium alone (no treatment). In addition to these two situations, results of 3 other experimental conditions are shown: 1) PBMCs treated with RM-4848 before being infected (pre-infection); 2) RM-4848 present during the 24 hours infection period (during infection) and then washed away; 3) RM-4848 added 24 hours after infection (post infection). Mean values ± standard errors are shown.
    Pbmc Isolation, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 93/100, based on 179 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pbmc isolation/product/Becton Dickinson
    Average 93 stars, based on 179 article reviews
    Price from $9.99 to $1999.99
    pbmc isolation - by Bioz Stars, 2020-09
    93/100 stars
      Buy from Supplier

    92
    PAA Laboratories peripheral blood mononuclear cells pbmc
    ( a ) p24 viral antigen levels 10 days post-infection in the supernatant of <t>HIV-1</t> infected PBMCs treated with TIZ (10 μg/mL) or RM-4848 (10 μg/mL). Results obtained with <t>PBMC</t> of 20 different healthy donors are shown. Median values and statistical significance is presented. ( b ) Inhibition (%) of viral replication in PBMC of 20 different healthy donors infected in vitro with HIV-1 in the presence of TIZ- (10 μg/mL) or RM-4848- (10 μg/mL). Median values and statistical significance is presented. ( c ) p24 viral antigen levels in PBMCs infected with HIV-1 in the presence of RM-4848 throughout infection (RM-4848)(10 μg/mL) or in medium alone (no treatment). In addition to these two situations, results of 3 other experimental conditions are shown: 1) PBMCs treated with RM-4848 before being infected (pre-infection); 2) RM-4848 present during the 24 hours infection period (during infection) and then washed away; 3) RM-4848 added 24 hours after infection (post infection). Mean values ± standard errors are shown.
    Peripheral Blood Mononuclear Cells Pbmc, supplied by PAA Laboratories, used in various techniques. Bioz Stars score: 92/100, based on 155 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/peripheral blood mononuclear cells pbmc/product/PAA Laboratories
    Average 92 stars, based on 155 article reviews
    Price from $9.99 to $1999.99
    peripheral blood mononuclear cells pbmc - by Bioz Stars, 2020-09
    92/100 stars
      Buy from Supplier

    93
    Greiner Bio peripheral blood mononuclear cells pbmcs
    ( a ) p24 viral antigen levels 10 days post-infection in the supernatant of <t>HIV-1</t> infected PBMCs treated with TIZ (10 μg/mL) or RM-4848 (10 μg/mL). Results obtained with <t>PBMC</t> of 20 different healthy donors are shown. Median values and statistical significance is presented. ( b ) Inhibition (%) of viral replication in PBMC of 20 different healthy donors infected in vitro with HIV-1 in the presence of TIZ- (10 μg/mL) or RM-4848- (10 μg/mL). Median values and statistical significance is presented. ( c ) p24 viral antigen levels in PBMCs infected with HIV-1 in the presence of RM-4848 throughout infection (RM-4848)(10 μg/mL) or in medium alone (no treatment). In addition to these two situations, results of 3 other experimental conditions are shown: 1) PBMCs treated with RM-4848 before being infected (pre-infection); 2) RM-4848 present during the 24 hours infection period (during infection) and then washed away; 3) RM-4848 added 24 hours after infection (post infection). Mean values ± standard errors are shown.
    Peripheral Blood Mononuclear Cells Pbmcs, supplied by Greiner Bio, used in various techniques. Bioz Stars score: 93/100, based on 230 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/peripheral blood mononuclear cells pbmcs/product/Greiner Bio
    Average 93 stars, based on 230 article reviews
    Price from $9.99 to $1999.99
    peripheral blood mononuclear cells pbmcs - by Bioz Stars, 2020-09
    93/100 stars
      Buy from Supplier

    91
    Becton Dickinson flow cytometry pbmcs
    Rituximab ( RTX ) leads to further phenotypical changes in CD56 dim natural killer ( NK ) cells. Freshly isolated peripheral blood mononuclear cells <t>(PBMCs)</t> were incubated with (+) or without ( w/o ) RTX overnight. The next day PBMCs were stained with a mixture of antibodies (anti-CD3 PE-Dazzle, anti-CD56 Bv421 and anti-CD16 FITC and CD69/CD137 PE) and analyzed by flow <t>cytometry.</t> NK cells were identified as CD3-CD56+ cells in the lymphocyte gate and fluorescence intensity of PE on CD56 dim NK cells was measured. a CD69 PE. Histogram shows one representative donor; gray incubation + RTX, black w/o RTX. Left diagram mean CD69 PE fluorescence intensity in samples from 13 donors. Statistical significance was determined with the Wilcoxon signed rank test (p = 0.0002). Right diagram Δ % in CD56 dim NK was determined by (CD69-positive CD56 dim NK cells after incubation + RTX) - (CD69-positive CD56 dim NK cells after incubation w/o RTX); bar median. b CD137(41BB)-PE; same donor, layout and abbreviations as in a ; n = 11 donors; p = 0.001
    Flow Cytometry Pbmcs, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 91/100, based on 254 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/flow cytometry pbmcs/product/Becton Dickinson
    Average 91 stars, based on 254 article reviews
    Price from $9.99 to $1999.99
    flow cytometry pbmcs - by Bioz Stars, 2020-09
    91/100 stars
      Buy from Supplier

    93
    Axis-Shield Diagnostics blood mononuclear cells pbmcs
    Inhibition of cytokine secretion in LPS-stimulated <t>PBMCs</t> by <t>rhIL-1ra.</t> LPS, lipopolysaccharide; PBMCs, peripheral blood mononuclear cells; rh, human recombinant; IL, interleukin; ra, receptor antagonist; low, LPS + 125 ng/ml rhIL-1ra; middle, LPS + 250 ng/ml rhIL-1ra; high, LPS + 500 ng/ml rhIL-1ra; IFN, interferon; TNF, tumor necrosis factor.
    Blood Mononuclear Cells Pbmcs, supplied by Axis-Shield Diagnostics, used in various techniques. Bioz Stars score: 93/100, based on 126 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/blood mononuclear cells pbmcs/product/Axis-Shield Diagnostics
    Average 93 stars, based on 126 article reviews
    Price from $9.99 to $1999.99
    blood mononuclear cells pbmcs - by Bioz Stars, 2020-09
    93/100 stars
      Buy from Supplier

    Image Search Results


    ShK-186 inhibits antineutrophil cytoplasmic antibodies targeting proteinase 3 (PR3-ANCA) production in vitro . (A) IgG production after peripheral blood mononuclear cell (PBMC) stimulation with CpG, B cell-activating factor (BAFF), and interleukin (IL)-21 from 5 healthy controls (HCs) (open circles) and 13 granulomatosis with polyangiitis (GPA) patients (gray circles) in the presence and absence of 1 nM ShK-186. (B) PR3-ANCA production after PBMC stimulation with CpG, BAFF, and IL-21 from 13 GPA patients in the presence and absence of 1 nM ShK-186 (* p

    Journal: Frontiers in Immunology

    Article Title: Kv1.3 Channel Blockade Modulates the Effector Function of B Cells in Granulomatosis with Polyangiitis

    doi: 10.3389/fimmu.2017.01205

    Figure Lengend Snippet: ShK-186 inhibits antineutrophil cytoplasmic antibodies targeting proteinase 3 (PR3-ANCA) production in vitro . (A) IgG production after peripheral blood mononuclear cell (PBMC) stimulation with CpG, B cell-activating factor (BAFF), and interleukin (IL)-21 from 5 healthy controls (HCs) (open circles) and 13 granulomatosis with polyangiitis (GPA) patients (gray circles) in the presence and absence of 1 nM ShK-186. (B) PR3-ANCA production after PBMC stimulation with CpG, BAFF, and IL-21 from 13 GPA patients in the presence and absence of 1 nM ShK-186 (* p

    Article Snippet: Briefly, peripheral blood mononuclear cells (PBMCs) were isolated from GPA patients, who produce ANCA upon in vitro induction , and stored in RPMI 1640 (Lonza, Basel, Switzerland) supplemented with 50 µg/mL gentamycin (GIBCO, Life Technologies, Grand Island, NY, USA), 10% fetal calf serum (FCS, Lonza), and 10% dimethyl sulfoxide (DMSO).

    Techniques: In Vitro

    Detection of ApoE isoforms in differentiating hNP1 cells. Differentiating hNP1 cells were exposed to medium alone (Unt), mock-infected PBMC supernatants (Mock), and HIV-infected PBMC supernatants (HIV), and to rApoE3 or rApoE4 at 3 μg/ml, as described in Materials and Methods. At day 16 of differentiation, and 24 h after replenishment of rApoE in the cultures, cells were lysed for quantitative immunoblotting to detect ApoE. Lower blot depicts representative endogenous ApoE detected in cell culture lysates with molecular weight of 34 kDa, and rApoE with molecular weight of 36 kDa. Upper blot corresponds to beta-actin used as an internal control. For comparison, recombinant ApoE3 and ApoE4 were loaded directly on the gel at a 3 ng per lane, as shown. Bar graphs present ApoE signal normalized to beta-actin; each bar represents the mean of two experiments

    Journal: Journal of Neuroimmune Pharmacology

    Article Title: Apolipoprotein E4 Suppresses Neuronal-Specific Gene Expression in Maturing Neuronal Progenitor Cells Exposed to HIV

    doi: 10.1007/s11481-017-9734-9

    Figure Lengend Snippet: Detection of ApoE isoforms in differentiating hNP1 cells. Differentiating hNP1 cells were exposed to medium alone (Unt), mock-infected PBMC supernatants (Mock), and HIV-infected PBMC supernatants (HIV), and to rApoE3 or rApoE4 at 3 μg/ml, as described in Materials and Methods. At day 16 of differentiation, and 24 h after replenishment of rApoE in the cultures, cells were lysed for quantitative immunoblotting to detect ApoE. Lower blot depicts representative endogenous ApoE detected in cell culture lysates with molecular weight of 34 kDa, and rApoE with molecular weight of 36 kDa. Upper blot corresponds to beta-actin used as an internal control. For comparison, recombinant ApoE3 and ApoE4 were loaded directly on the gel at a 3 ng per lane, as shown. Bar graphs present ApoE signal normalized to beta-actin; each bar represents the mean of two experiments

    Article Snippet: “HIV-Exposed” HIV-1 containing supernatants were obtained from human peripheral blood mononuclear cells (PBMC) that were stimulated with mitogens phytohemagglutinin (PHA, Sigma-Aldrich, St Lois, MO) and recombinant human interleukin-2 (IL-2, Roche Diagnostics, Indianapolis, IN), then infected with HIV-1 as described previously (McCarthy et al. ).

    Techniques: Infection, Cell Culture, Molecular Weight, Recombinant

    Comparison of HBV-specific cell responses in the presence or absence of HBeAg. (A) C protein-specific T-cell responses elicited in ELISPOT assay; (B) Cytokine IFN-Y responses in cultured supernatants of PBMCs stimulated by C protein. HBeAg(−): HBeAg-negative subjects; HBeAg(+): HBeAg-positive subjects; HIV+: HIV co-infected; HIV-: not co-infected with HIV.

    Journal: Virology Journal

    Article Title: Hepatitis B virus (HBV)-specific T-cell responses to recombinant HBV core protein in patients with normal liver function and co-infected with chronic HBV and human immunodeficiency virus 1 (HIV-1)

    doi: 10.1186/1743-422X-10-232

    Figure Lengend Snippet: Comparison of HBV-specific cell responses in the presence or absence of HBeAg. (A) C protein-specific T-cell responses elicited in ELISPOT assay; (B) Cytokine IFN-Y responses in cultured supernatants of PBMCs stimulated by C protein. HBeAg(−): HBeAg-negative subjects; HBeAg(+): HBeAg-positive subjects; HIV+: HIV co-infected; HIV-: not co-infected with HIV.

    Article Snippet: Human interferon (IFN)-γ ELISPOT assay Peripheral whole blood was obtained from all subjects and peripheral blood mononuclear cells (PBMCs) were isolated by density gradient centrifugation with Ficoll Lymphoprep (Axis –Shield PoC AS, Oslo, Norway).

    Techniques: Enzyme-linked Immunospot, Cell Culture, Infection

    The distribution of fragments of mRNA, rRNA, tRNA, miRNA, and others in the small RNA libraries. a The PBMC of heat-stressed cattle. b The PBMC of non-heat-stressed cattle

    Journal: Cell Stress & Chaperones

    Article Title: Differential expression of microRNAs associated with thermal stress in Frieswal (Bos taurus x Bos indicus) crossbred dairy cattle

    doi: 10.1007/s12192-017-0833-6

    Figure Lengend Snippet: The distribution of fragments of mRNA, rRNA, tRNA, miRNA, and others in the small RNA libraries. a The PBMC of heat-stressed cattle. b The PBMC of non-heat-stressed cattle

    Article Snippet: To identify miRNAs in Frieswal cattle, two independent small RNA libraries were constructed from the PBMC of normal and heat-stressed animals through Ion Torrent deep sequencing.

    Techniques:

    Fold changes of HSP70 and HSP90 in the Frieswal PBMC collected 18 °C (normal) and 45 °C (heat stress) environmental temperatures. * P

    Journal: Cell Stress & Chaperones

    Article Title: Differential expression of microRNAs associated with thermal stress in Frieswal (Bos taurus x Bos indicus) crossbred dairy cattle

    doi: 10.1007/s12192-017-0833-6

    Figure Lengend Snippet: Fold changes of HSP70 and HSP90 in the Frieswal PBMC collected 18 °C (normal) and 45 °C (heat stress) environmental temperatures. * P

    Article Snippet: To identify miRNAs in Frieswal cattle, two independent small RNA libraries were constructed from the PBMC of normal and heat-stressed animals through Ion Torrent deep sequencing.

    Techniques:

    Systemic immune response to LCMV-GP33 peptide C57BL/6 mice (5 mice/group) were challenged subcutaneously with TC-1 tumor cells and subsequently treated with various combinations of cisplatin, CpG and GP33 peptide as shown. The PBMCs were analyzed 1 week after the last antigen delivery. The presence of GP33-specific CD8+ T cells in PBMCs was analyzed using intracellular cytokine staining for IFN-γ and CD8+ staining followed by flow cytometry analysis. A. Representative flow cytometry contour plot depicting the frequency of IFN-γ-secreting CD8+ T cells among PBMCs after being pulsed with GP33 peptide. B. Bar graph depicting the number of IFN-γ+ CD8+ T cells among PBMCs (mean ±S.E).

    Journal: Vaccine

    Article Title: Cancer immunotherapy using a potent immunodominant CTL epitope

    doi: 10.1016/j.vaccine.2014.09.021

    Figure Lengend Snippet: Systemic immune response to LCMV-GP33 peptide C57BL/6 mice (5 mice/group) were challenged subcutaneously with TC-1 tumor cells and subsequently treated with various combinations of cisplatin, CpG and GP33 peptide as shown. The PBMCs were analyzed 1 week after the last antigen delivery. The presence of GP33-specific CD8+ T cells in PBMCs was analyzed using intracellular cytokine staining for IFN-γ and CD8+ staining followed by flow cytometry analysis. A. Representative flow cytometry contour plot depicting the frequency of IFN-γ-secreting CD8+ T cells among PBMCs after being pulsed with GP33 peptide. B. Bar graph depicting the number of IFN-γ+ CD8+ T cells among PBMCs (mean ±S.E).

    Article Snippet: To detect HPV16 E7-specific and LCMV GP33-specific CD8+ T cell responses by IFN-γ intracellular staining, peripheral blood mononuclear cells (PBMCs), single-cell suspended splenocytes or tumor infiltrating lymphocytes were stimulated with either HPV E7aa49-57 or LCMV GP33 peptide (1μg/ml) in the presence of Golgiplug (BD Pharmingen, San Diego, CA) at 37°C overnight.

    Techniques: Mouse Assay, Staining, Flow Cytometry, Cytometry

    Proliferation of lymphocytes in LNs and PBMCs as measured by flow cytometry. (A) Ki-67 immunostaining of lymphocytes isolated from LNs of healthy animals on day −18 preexposure ( n = 2) and from LNs of surviving animals ( n = 4) on days 9 and 22 p.i. (B to D) Ki-67 immunostaining of CD4 + T cells (B), CD8 + T cells (C), and CD20 + B cells (D) in PBMCs in treated and untreated survivors versus moribund NHPs. Abbreviations: LN, lymph node; NHP, nonhuman primate; PBMCs, peripheral blood mononuclear cells; p.i., postinfection.

    Journal: Journal of Virology

    Article Title: [18F]-Fluorodeoxyglucose Uptake in Lymphoid Tissue Serves as a Predictor of Disease Outcome in the Nonhuman Primate Model of Monkeypox Virus Infection

    doi: 10.1128/JVI.00897-17

    Figure Lengend Snippet: Proliferation of lymphocytes in LNs and PBMCs as measured by flow cytometry. (A) Ki-67 immunostaining of lymphocytes isolated from LNs of healthy animals on day −18 preexposure ( n = 2) and from LNs of surviving animals ( n = 4) on days 9 and 22 p.i. (B to D) Ki-67 immunostaining of CD4 + T cells (B), CD8 + T cells (C), and CD20 + B cells (D) in PBMCs in treated and untreated survivors versus moribund NHPs. Abbreviations: LN, lymph node; NHP, nonhuman primate; PBMCs, peripheral blood mononuclear cells; p.i., postinfection.

    Article Snippet: PBMC samples and lymphocytes isolated from LNs were surface stained with a custom antibody cocktail (BD Biosciences) for CD3, CD4, CD8, CD14, CD16, CD20, CD45, and Ki-67.

    Techniques: Flow Cytometry, Cytometry, Immunostaining, Isolation

    Dose–response curves for ADCC. The procedures for OxLDL incubated and Ab treatment were the same as described in the legend for Fig. . As effectors normal rat PBMC were added to the culture instead of complement. Note increased 51Cr release

    Journal: Cell Stress & Chaperones

    Article Title: Evidence of a role for both anti-Hsp70 antibody and endothelial surface membrane Hsp70 in atherosclerosis

    doi: 10.1007/s12192-013-0404-4

    Figure Lengend Snippet: Dose–response curves for ADCC. The procedures for OxLDL incubated and Ab treatment were the same as described in the legend for Fig. . As effectors normal rat PBMC were added to the culture instead of complement. Note increased 51Cr release

    Article Snippet: To determine antibody-dependent cellular cytotoxicity (ADCC), peripheral blood mononuclear cells (PBMC) were isolated from healthy rats by density centrifugation (Lymphoprep, density 1,083; Nycomed Pharmaceuticals Oslo, Norway) as described previously (Jurgens et al. ).

    Techniques: Incubation

    miR-20b expression in T cells and in PBMC from natalizumab-associated progressive multifocal leukoencephalopathy (PML). (A) PBMC were isolated using a Ficoll gradient and CD4+ T cells were sorted with magnetic beads ( n = 9; randomly selected patients of our initial longitudinal cohort consisting of 17 patients). miR-20b expression (qPCR) was measured in the CD4+ and CD4-negative fraction and compared using t- test. (B) CD4+ cells were activated by plate-bound anti-CD3-antibody and soluble anti-CD28-antibody without further cytokines, termed “Th0”, and with IL-1 (10 ng/mL), IL-6 (10 ng/mL), IL-23 (10 ng/mL) and TGF- β (5 ng/mL), termed “Th17”, as well as freshly isolated CD4+ T cells, termed “fresh”. miR-20b levels were measured by qPCR. (C) miR-20b levels in PBMC from natalizumab-treated patients ( n = 9, left bar) were compared to miR-20b levels in two PBMC samples from patients with natalizumab-associated PML (each patient one bar).

    Journal: Annals of Clinical and Translational Neurology

    Article Title: Natalizumab restores aberrant miRNA expression profile in multiple sclerosis and reveals a critical role for miR-20b

    doi: 10.1002/acn3.152

    Figure Lengend Snippet: miR-20b expression in T cells and in PBMC from natalizumab-associated progressive multifocal leukoencephalopathy (PML). (A) PBMC were isolated using a Ficoll gradient and CD4+ T cells were sorted with magnetic beads ( n = 9; randomly selected patients of our initial longitudinal cohort consisting of 17 patients). miR-20b expression (qPCR) was measured in the CD4+ and CD4-negative fraction and compared using t- test. (B) CD4+ cells were activated by plate-bound anti-CD3-antibody and soluble anti-CD28-antibody without further cytokines, termed “Th0”, and with IL-1 (10 ng/mL), IL-6 (10 ng/mL), IL-23 (10 ng/mL) and TGF- β (5 ng/mL), termed “Th17”, as well as freshly isolated CD4+ T cells, termed “fresh”. miR-20b levels were measured by qPCR. (C) miR-20b levels in PBMC from natalizumab-treated patients ( n = 9, left bar) were compared to miR-20b levels in two PBMC samples from patients with natalizumab-associated PML (each patient one bar).

    Article Snippet: Peripheral blood mononuclear cells (PBMC) were isolated from whole blood by Ficoll (Biochrom, Berlin, Germany) gradient and CD4+ cells were isolated by magnetic bead separation using STEMCELL EasySep Human CD4+ T Cell Enrichment Kit according to the manufacturer’s instructions.

    Techniques: Expressing, Isolation, Magnetic Beads, Real-time Polymerase Chain Reaction

    Semi-quantitative PCR analysis of BLV pol detects differences in proviral load between BLV SGV and BLV-rabbits. PBMC were harvested at month indicated after challenge with 1 × 10 8 FLK/BLV cells from indicated rabbits that had been infected with BLV SGV for 24 months or from age-matched naïve controls. DNA from 50,000 PMBC was subjected to nested PCR and gel electrophoresis and stained with ethidium bromide. The position of the 591-bp amplicon is designated by the labeled arrow. In parallel, DNA from 20,000 PBMC was subjected to PCR with primers complementary to β- actin . The position of the 594-bp amplicon is designated by the labeled arrow and the signals were quantified by densitometry.

    Journal: Virology

    Article Title: Long-term infection with retroviral structural gene vector provides protection against bovine leukemia virus disease in rabbits

    doi: 10.1016/j.virol.2004.09.001

    Figure Lengend Snippet: Semi-quantitative PCR analysis of BLV pol detects differences in proviral load between BLV SGV and BLV-rabbits. PBMC were harvested at month indicated after challenge with 1 × 10 8 FLK/BLV cells from indicated rabbits that had been infected with BLV SGV for 24 months or from age-matched naïve controls. DNA from 50,000 PMBC was subjected to nested PCR and gel electrophoresis and stained with ethidium bromide. The position of the 591-bp amplicon is designated by the labeled arrow. In parallel, DNA from 20,000 PBMC was subjected to PCR with primers complementary to β- actin . The position of the 594-bp amplicon is designated by the labeled arrow and the signals were quantified by densitometry.

    Article Snippet: To prepare peripheral blood mononuclear cell (PBMC) samples for PCR, PBMCs were isolated by a Ficoll-Paque (Pharmacia) gradient centrifugation and washed three times with phosphate buffered saline.

    Techniques: Real-time Polymerase Chain Reaction, Infection, Nested PCR, Nucleic Acid Electrophoresis, Staining, Amplification, Labeling, Polymerase Chain Reaction

    Semiquantitative PCR analysis to evaluate differences in proviral load. (A) PCR to detect BLV pol in PBMC from treated rabbits. Rabbit PBMC were harvested at 1, 4, and 10 months postinoculation and subjected to PCR with BLV pol primers KB560 and KB561 (10 × 10 4 PBMC) or β- actin to control for sample variation (2 × 10 4 PBMC). Each panel is labeled with month of sample harvest. Lanes are labeled with the source of PBMC DNA by rabbit number and treatment, FLK(BLV) (positive control DNA [100 ng] from BLV-producing fetal lamb kidney cells), or marker (pGem DNA size standard [Promega]). In lower panels, the corresponding rabbit samples are designated by the matching parallel black lines. Positions of BLV pol amplicon (591 bp) and β- actin amplicon (594 bp) are designated. *, not determined (ND) because sample was not available; †, not determined because the animal died before harvest. (B) PCR standard curve. PBMC lysate harvested at 1 month postinoculation from BLV rabbit 44-10 was serially diluted in a range of 5 × 10 2 to 5 × 10 4 PBMC and subjected to PCR with BLV pol primers KB560 and KB561. Each lane is labeled with the number of cells used for PCR amplification, FLK(BLV) (positive control DNA [100 ng] from BLV-producing fetal lamb kidney cells), or marker (pGem DNA size standard [Promega]). The arrow on the left indicates the position of BLV pol amplicon (591 bp), and the lines on the right indicate positions of 676- and 571-bp DNA size markers.

    Journal: Journal of Virology

    Article Title: Bovine Leukemia Virus Structural Gene Vectors Are Immunogenic and Lack Pathogenicity in a Rabbit Model

    doi:

    Figure Lengend Snippet: Semiquantitative PCR analysis to evaluate differences in proviral load. (A) PCR to detect BLV pol in PBMC from treated rabbits. Rabbit PBMC were harvested at 1, 4, and 10 months postinoculation and subjected to PCR with BLV pol primers KB560 and KB561 (10 × 10 4 PBMC) or β- actin to control for sample variation (2 × 10 4 PBMC). Each panel is labeled with month of sample harvest. Lanes are labeled with the source of PBMC DNA by rabbit number and treatment, FLK(BLV) (positive control DNA [100 ng] from BLV-producing fetal lamb kidney cells), or marker (pGem DNA size standard [Promega]). In lower panels, the corresponding rabbit samples are designated by the matching parallel black lines. Positions of BLV pol amplicon (591 bp) and β- actin amplicon (594 bp) are designated. *, not determined (ND) because sample was not available; †, not determined because the animal died before harvest. (B) PCR standard curve. PBMC lysate harvested at 1 month postinoculation from BLV rabbit 44-10 was serially diluted in a range of 5 × 10 2 to 5 × 10 4 PBMC and subjected to PCR with BLV pol primers KB560 and KB561. Each lane is labeled with the number of cells used for PCR amplification, FLK(BLV) (positive control DNA [100 ng] from BLV-producing fetal lamb kidney cells), or marker (pGem DNA size standard [Promega]). The arrow on the left indicates the position of BLV pol amplicon (591 bp), and the lines on the right indicate positions of 676- and 571-bp DNA size markers.

    Article Snippet: To prepare PBMCs samples for PCR, PBMCs were isolated by Ficoll-Paque (Pharmacia) gradient centrifugation and washed three times with phosphate-buffered saline.

    Techniques: Polymerase Chain Reaction, Labeling, Positive Control, Marker, Amplification

    Generation, isolation, and characterization of epidermal growth factor receptor (EGFR)‐specific chimeric antigen receptor (CAR)‐engineered natural killer (NK) cells (EGFR‐CAR NK cells). (A) Flow cytometric analysis of phenotypic and subset composition of peripheral blood mononuclear cells (PBMCs) labeled with anti‐CD3‐PE‐Cy7, anti‐CD56‐PE. (B‐C) Flow cytometric analysis of phenotypic and subset composition of NK cells labeled with anti‐CD3‐PE‐Cy7, anti‐CD56‐PE, and anti‐CD69‐APC‐Cy7. (D) The percentage of CD3‐/CD56 + cells in day 0 PBMCs and day14 PBMC culture. (E) Real‐time PCR and (F) Western blotting analyses of the expression of exogenous CD3ζ in the non‐transduced NK cells, Con‐CAR NK cells, EGFR‐CAR‐1 NK cells, and EGFR‐CAR‐2 NK cells. β‐actin was used as an endogenous control. (G) The transduced NK cells stained with IgG‐FITC and EGFR‐FITC antibodies were detected by flow cytometry

    Journal: Cell Proliferation

    Article Title: Targeting epidermal growth factor‐overexpressing triple‐negative breast cancer by natural killer cells expressing a specific chimeric antigen receptor, et al. Targeting epidermal growth factor‐overexpressing triple‐negative breast cancer by natural killer cells expressing a specific chimeric antigen receptor

    doi: 10.1111/cpr.12858

    Figure Lengend Snippet: Generation, isolation, and characterization of epidermal growth factor receptor (EGFR)‐specific chimeric antigen receptor (CAR)‐engineered natural killer (NK) cells (EGFR‐CAR NK cells). (A) Flow cytometric analysis of phenotypic and subset composition of peripheral blood mononuclear cells (PBMCs) labeled with anti‐CD3‐PE‐Cy7, anti‐CD56‐PE. (B‐C) Flow cytometric analysis of phenotypic and subset composition of NK cells labeled with anti‐CD3‐PE‐Cy7, anti‐CD56‐PE, and anti‐CD69‐APC‐Cy7. (D) The percentage of CD3‐/CD56 + cells in day 0 PBMCs and day14 PBMC culture. (E) Real‐time PCR and (F) Western blotting analyses of the expression of exogenous CD3ζ in the non‐transduced NK cells, Con‐CAR NK cells, EGFR‐CAR‐1 NK cells, and EGFR‐CAR‐2 NK cells. β‐actin was used as an endogenous control. (G) The transduced NK cells stained with IgG‐FITC and EGFR‐FITC antibodies were detected by flow cytometry

    Article Snippet: Peripheral blood mononuclear cells were isolated from the whole blood of healthy donors by Ficoll density gradient centrifugation.

    Techniques: Isolation, Labeling, Real-time Polymerase Chain Reaction, Western Blot, Expressing, Staining, Flow Cytometry

    EAT-2 over-expression enhances human DC maturation. Human PBMCs were isolated from fresh buffy coat material and were either mock infected or infected with MOI of 10000 with rAd5-EAT2 or rAd5-Null. At 72 hpi, cells were stained for surface expression of CD86 (A), HLA-DR (B), CD80 (C) and CD83 (D) and FACS analysis was performed. Data are representative of at least three independent experiments from three different blood donors with similar results. Samples were plated in quadruplicates. Bars represent mean ± SD. Statistical analysis was completed using a one way ANOVA with a Student–Newman–Keuls post-hoc test. P

    Journal: International Immunology

    Article Title: Manipulation of EAT-2 expression promotes induction of multiple beneficial regulatory and effector functions of the human innate immune system as a novel immunomodulatory strategy

    doi: 10.1093/intimm/dxt061

    Figure Lengend Snippet: EAT-2 over-expression enhances human DC maturation. Human PBMCs were isolated from fresh buffy coat material and were either mock infected or infected with MOI of 10000 with rAd5-EAT2 or rAd5-Null. At 72 hpi, cells were stained for surface expression of CD86 (A), HLA-DR (B), CD80 (C) and CD83 (D) and FACS analysis was performed. Data are representative of at least three independent experiments from three different blood donors with similar results. Samples were plated in quadruplicates. Bars represent mean ± SD. Statistical analysis was completed using a one way ANOVA with a Student–Newman–Keuls post-hoc test. P

    Article Snippet: To identify human immune cells transduced by Ad-mediated EAT-2 or GFP gene transduction, human PBMCs (2×106 ) were infected with multiplicity of infection (MOI) of 10000 of Ad5-EAT-2 or Ad5-GFP for 48h and were then stained with the following antibodies: APC-Cy7-CD14, PE-CD1a, Alexa Fluor 700-CD8a, V450-CD4, APC-CD65 and PerCpCy5.5-CD3 (all at 4 µg ml−1 ; BD Biosciences).

    Techniques: Over Expression, Isolation, Infection, Staining, Expressing, FACS

    Transduction of human innate and adaptive immune cells by GFP-expressing rAd5 vector. Human PBMCs were isolated from fresh buffy coat material and were either mock infected or infected with MOI of 10000 with rAd5-GFP. FACS analysis for GFP expression at 48h following rAd5-GFP infection of human NK cells (A), CD4 + and CD8 + T cells (B), CD14 + monocytes and CD1a + cells (C) is shown. Data were collected using a BD LSRII flow cytometer and analyzed using FlowJo software.

    Journal: International Immunology

    Article Title: Manipulation of EAT-2 expression promotes induction of multiple beneficial regulatory and effector functions of the human innate immune system as a novel immunomodulatory strategy

    doi: 10.1093/intimm/dxt061

    Figure Lengend Snippet: Transduction of human innate and adaptive immune cells by GFP-expressing rAd5 vector. Human PBMCs were isolated from fresh buffy coat material and were either mock infected or infected with MOI of 10000 with rAd5-GFP. FACS analysis for GFP expression at 48h following rAd5-GFP infection of human NK cells (A), CD4 + and CD8 + T cells (B), CD14 + monocytes and CD1a + cells (C) is shown. Data were collected using a BD LSRII flow cytometer and analyzed using FlowJo software.

    Article Snippet: To identify human immune cells transduced by Ad-mediated EAT-2 or GFP gene transduction, human PBMCs (2×106 ) were infected with multiplicity of infection (MOI) of 10000 of Ad5-EAT-2 or Ad5-GFP for 48h and were then stained with the following antibodies: APC-Cy7-CD14, PE-CD1a, Alexa Fluor 700-CD8a, V450-CD4, APC-CD65 and PerCpCy5.5-CD3 (all at 4 µg ml−1 ; BD Biosciences).

    Techniques: Transduction, Expressing, Plasmid Preparation, Isolation, Infection, FACS, Flow Cytometry, Cytometry, Software

    Intracellular staining analysis for EAT-2 expression in human immune cells following rAd5-EAT2 infection. Human PBMCs were isolated from fresh buffy coat material and were either mock infected or infected with MOI of 10000 with rAd5-EAT2 or rAd5-EAT2(R31Q). Intracellular staining analysis for EAT-2 expression at 48h following Ad infection of human NK cells (A), CD14 + monocytes (B) and CD1a + DCs (C) is shown. Data were collected using a BD LSRII flow cytometer and analyzed using FlowJo software.

    Journal: International Immunology

    Article Title: Manipulation of EAT-2 expression promotes induction of multiple beneficial regulatory and effector functions of the human innate immune system as a novel immunomodulatory strategy

    doi: 10.1093/intimm/dxt061

    Figure Lengend Snippet: Intracellular staining analysis for EAT-2 expression in human immune cells following rAd5-EAT2 infection. Human PBMCs were isolated from fresh buffy coat material and were either mock infected or infected with MOI of 10000 with rAd5-EAT2 or rAd5-EAT2(R31Q). Intracellular staining analysis for EAT-2 expression at 48h following Ad infection of human NK cells (A), CD14 + monocytes (B) and CD1a + DCs (C) is shown. Data were collected using a BD LSRII flow cytometer and analyzed using FlowJo software.

    Article Snippet: To identify human immune cells transduced by Ad-mediated EAT-2 or GFP gene transduction, human PBMCs (2×106 ) were infected with multiplicity of infection (MOI) of 10000 of Ad5-EAT-2 or Ad5-GFP for 48h and were then stained with the following antibodies: APC-Cy7-CD14, PE-CD1a, Alexa Fluor 700-CD8a, V450-CD4, APC-CD65 and PerCpCy5.5-CD3 (all at 4 µg ml−1 ; BD Biosciences).

    Techniques: Staining, Expressing, Infection, Isolation, Flow Cytometry, Cytometry, Software

    EAT-2 over-expression enhances the cytolytic activity of human NK cells. Human PBMCs isolated from fresh buffy coat material were either mock infected or infected with MOI of 10000 with rAd5-EAT2, rAd5-EAT2(R31Q) or rAd5-Null for 72h and then co-cultured with CFSE-labeled K562 cells for an additional 48h at an E:T ratio of 1:10. Viability of K562 cells was assessed by either PI (A and B) or ViViD (C). Data are representative of two (A) or three (B) independent experiments from three different blood donors with similar results. Samples were plated in quadruplicates. Bars represent mean ± SD. Statistical analysis was completed using a one way ANOVA with a Student–Newman–Keuls post-hoc test. P

    Journal: International Immunology

    Article Title: Manipulation of EAT-2 expression promotes induction of multiple beneficial regulatory and effector functions of the human innate immune system as a novel immunomodulatory strategy

    doi: 10.1093/intimm/dxt061

    Figure Lengend Snippet: EAT-2 over-expression enhances the cytolytic activity of human NK cells. Human PBMCs isolated from fresh buffy coat material were either mock infected or infected with MOI of 10000 with rAd5-EAT2, rAd5-EAT2(R31Q) or rAd5-Null for 72h and then co-cultured with CFSE-labeled K562 cells for an additional 48h at an E:T ratio of 1:10. Viability of K562 cells was assessed by either PI (A and B) or ViViD (C). Data are representative of two (A) or three (B) independent experiments from three different blood donors with similar results. Samples were plated in quadruplicates. Bars represent mean ± SD. Statistical analysis was completed using a one way ANOVA with a Student–Newman–Keuls post-hoc test. P

    Article Snippet: To identify human immune cells transduced by Ad-mediated EAT-2 or GFP gene transduction, human PBMCs (2×106 ) were infected with multiplicity of infection (MOI) of 10000 of Ad5-EAT-2 or Ad5-GFP for 48h and were then stained with the following antibodies: APC-Cy7-CD14, PE-CD1a, Alexa Fluor 700-CD8a, V450-CD4, APC-CD65 and PerCpCy5.5-CD3 (all at 4 µg ml−1 ; BD Biosciences).

    Techniques: Over Expression, Activity Assay, Isolation, Infection, Cell Culture, Labeling

    Intracellular and plasma pharmacokinetic profiles of imatinib in five individual patients (solid lines, plasma concentrations; broken lines, peripheral blood mononuclear cell concentrations)

    Journal: British Journal of Clinical Pharmacology

    Article Title: Population pharmacokinetics of imatinib and the role of ?1-acid glycoprotein

    doi: 10.1111/j.1365-2125.2006.02719.x

    Figure Lengend Snippet: Intracellular and plasma pharmacokinetic profiles of imatinib in five individual patients (solid lines, plasma concentrations; broken lines, peripheral blood mononuclear cell concentrations)

    Article Snippet: In these five patients, intracellular concentrations of imatinib were also measured in peripheral blood mononuclear cells (PBMC) obtained from four blood samples drawn before and 2, 4 and 6 h after drug intake using Vacutainer® CPT (Cell Preparation Tubes; Becton Dickinson, Allschwil, Switzerland), according to the manufacturer’s recommended procedure and the method previously developed in our laboratory for the intracellular measurement of anti-HIV drugs [ ].

    Techniques:

    Isolation and molecular characterization of T cell subsets from PNH patients ( A ) Gating strategy for T cell subsets for cell sorting. PBMCs were stained with ViViD, anti-CD14-Pacific Blue, anti-CD19-Pacific Blue, anti-CD3-BV605, anti-CD4-V500, anti-CD8-APC-H7, anti-CD45RA-PE-Cy7, and anti-CD45RO-APC. Lymphocytes were gated based on their scatter characteristics or forward scatter height vs. forward scatter area. Live T cells were gated based on positive for CD3 and negative for ViViD, CD14, and CD19 to remove dead cells, monocytes, and B cells. CD4 + and CD8 + T cells were then gated based on the characteristic expression patterns of CD45RA and CD45RO. Four different T cell subsets [CD4 + naïve (CD45RA + CD45RO - ), CD4 + memory (CD45RA - CD45RO + ), CD8 + naïve (CD45RA + CD45RO - ), and CD8 + memory (CD45RA - CD45RO + ) T cells] were sorted for RNA-seq. ( B ) Transcriptional validation of the naïve and memory T cell identity by analyzing expression levels of CD4, CD8, CCR5, and EOMES. FPKM (fragments per kilobase of exon per million fragments mapped). * p

    Journal: Journal of immunology (Baltimore, Md. : 1950)

    Article Title: T Cell Transcriptomes from Paroxysmal Nocturnal Hemoglobinuria Patients Reveal Novel Signaling Pathways

    doi: 10.4049/jimmunol.1601299

    Figure Lengend Snippet: Isolation and molecular characterization of T cell subsets from PNH patients ( A ) Gating strategy for T cell subsets for cell sorting. PBMCs were stained with ViViD, anti-CD14-Pacific Blue, anti-CD19-Pacific Blue, anti-CD3-BV605, anti-CD4-V500, anti-CD8-APC-H7, anti-CD45RA-PE-Cy7, and anti-CD45RO-APC. Lymphocytes were gated based on their scatter characteristics or forward scatter height vs. forward scatter area. Live T cells were gated based on positive for CD3 and negative for ViViD, CD14, and CD19 to remove dead cells, monocytes, and B cells. CD4 + and CD8 + T cells were then gated based on the characteristic expression patterns of CD45RA and CD45RO. Four different T cell subsets [CD4 + naïve (CD45RA + CD45RO - ), CD4 + memory (CD45RA - CD45RO + ), CD8 + naïve (CD45RA + CD45RO - ), and CD8 + memory (CD45RA - CD45RO + ) T cells] were sorted for RNA-seq. ( B ) Transcriptional validation of the naïve and memory T cell identity by analyzing expression levels of CD4, CD8, CCR5, and EOMES. FPKM (fragments per kilobase of exon per million fragments mapped). * p

    Article Snippet: For cell sorting, peripheral blood mononuclear cells (PBMCs) from PNH patients and healthy controls were separated from PB samples using Lymphocyte Separation Medium (MP Biomedicals, Santa Ana, CA).

    Techniques: Isolation, FACS, Staining, Expressing, RNA Sequencing Assay

    The systemic cellular immune response to sand fly saliva in volunteers in a cutaneous leishmaniasis-endemic area of Mali. Peripheral blood mononuclear cells (PBMCs) were obtained from 255 individuals aged 11–65 years from the villages of Kemena and Sougoula. IFN-γ, IL-12, IL-10, IL-13, and IL-5 levels were measured in the supernatants of PBMCs from exposed volunteers 96 hours after stimulation with Phlebotomus duboscqi salivary gland homogenate. ( a ) Overall cytokine production by tested individuals. ( b ) Pairwise correlation of expressed cytokines (log-transformed cytokine levels). Numbers in insets indicate the Spearman correlation coefficient and the 95% confidence intervals for each correlation. ( c ) Cytokine responses of individuals in ( a ) stratified by T H 1-predominant, T H 2-predominant, or T H 1/T H 2 mixed responses to sand fly salivary proteins.

    Journal: The Journal of Investigative Dermatology

    Article Title: Delayed-Type Hypersensitivity to Sand Fly Saliva in Humans from a Leishmaniasis-Endemic Area of Mali Is TH1-Mediated and Persists to Midlife

    doi: 10.1038/jid.2012.315

    Figure Lengend Snippet: The systemic cellular immune response to sand fly saliva in volunteers in a cutaneous leishmaniasis-endemic area of Mali. Peripheral blood mononuclear cells (PBMCs) were obtained from 255 individuals aged 11–65 years from the villages of Kemena and Sougoula. IFN-γ, IL-12, IL-10, IL-13, and IL-5 levels were measured in the supernatants of PBMCs from exposed volunteers 96 hours after stimulation with Phlebotomus duboscqi salivary gland homogenate. ( a ) Overall cytokine production by tested individuals. ( b ) Pairwise correlation of expressed cytokines (log-transformed cytokine levels). Numbers in insets indicate the Spearman correlation coefficient and the 95% confidence intervals for each correlation. ( c ) Cytokine responses of individuals in ( a ) stratified by T H 1-predominant, T H 2-predominant, or T H 1/T H 2 mixed responses to sand fly salivary proteins.

    Article Snippet: Blood collection and cytokine measurements To evaluate cytokine levels, PBMCs isolated by density-gradient centrifugation (Ficoll-Paque PLUS; GE Healthcare, Pittsburgh, PA) from blood collected in heparinized Vacutainer tubes (BD Diagnostics, Hunt Valley, MD).

    Techniques: Transformation Assay

    SV signaling activity is induced by influenza protein derived immune complexes. (A) Whole-blood and PBMCs prepared via Ficoll density centrifugation were stimulated with either PBS, SV, or R848 at 10ug/ml for 30mins prior to fixation and flow cytometric

    Journal: Vaccine

    Article Title: The Split Virus Influenza Vaccine rapidly activates immune cells through Fcγ Receptors

    doi: 10.1016/j.vaccine.2014.07.115

    Figure Lengend Snippet: SV signaling activity is induced by influenza protein derived immune complexes. (A) Whole-blood and PBMCs prepared via Ficoll density centrifugation were stimulated with either PBS, SV, or R848 at 10ug/ml for 30mins prior to fixation and flow cytometric

    Article Snippet: Peripheral blood mononuclear cells (PBMC) were obtained by density gradient centrifugation (Ficoll-Paque; GE healthcare) and cultured in RPMI 1640 + 10% FBS or AIM V media (Life Technologies).

    Techniques: Activity Assay, Derivative Assay, Centrifugation, Flow Cytometry

    Effects of PGRN on the differentiation of human OCs and PGRN as a predictor of osteoporosis. A , HBMCs and PBMCs were differentiated with RANKL in the absence or presence of PGRN. TRAP + MNCs were counted as described above. B , four sh-RNAs targeting human

    Journal: The Journal of Biological Chemistry

    Article Title: Progranulin and a Five Transmembrane Domain-Containing Receptor-like Gene Are the Key Components in Receptor Activator of Nuclear Factor κB (RANK)-dependent Formation of Multinucleated Osteoclasts *

    doi: 10.1074/jbc.M114.608786

    Figure Lengend Snippet: Effects of PGRN on the differentiation of human OCs and PGRN as a predictor of osteoporosis. A , HBMCs and PBMCs were differentiated with RANKL in the absence or presence of PGRN. TRAP + MNCs were counted as described above. B , four sh-RNAs targeting human

    Article Snippet: Human bone marrow cells (HBMCs) and peripheral blood mononuclear cells (PBMCs) were obtained from healthy volunteers and were separated by density gradient centrifugation using Ficoll-Histopaque (Sigma-Aldrich).

    Techniques:

    Nanoparticle formulation decreases curcumin cytotoxicity. SUPT1 cells (Panel A) or stimulated PBMCs (Panel B) were exposed to increasing concentrations (1, 5, 10, 25, 50 and 100 µM) of sol-curcumin, nano-curcumin, azidothymidine (AZT) or nano-apotransferrin (10, 50 and 100 µg) for 16 h, after which cell viability was determined by MTT assay. PBMCs were cultured in the presence of IL-2 (20 IU/ml). Cell viability in the absence of drug was defined as 0% cytotoxicity. Error bars indicate SD. ** P ≤0.01, and ***, P ≤0.001 compared to nano-curcumin. * indicates µg apotransferrin protein that carry equivalent molar concentration of the drug.

    Journal: PLoS ONE

    Article Title: Curcumin-Loaded Apotransferrin Nanoparticles Provide Efficient Cellular Uptake and Effectively Inhibit HIV-1 Replication In Vitro

    doi: 10.1371/journal.pone.0023388

    Figure Lengend Snippet: Nanoparticle formulation decreases curcumin cytotoxicity. SUPT1 cells (Panel A) or stimulated PBMCs (Panel B) were exposed to increasing concentrations (1, 5, 10, 25, 50 and 100 µM) of sol-curcumin, nano-curcumin, azidothymidine (AZT) or nano-apotransferrin (10, 50 and 100 µg) for 16 h, after which cell viability was determined by MTT assay. PBMCs were cultured in the presence of IL-2 (20 IU/ml). Cell viability in the absence of drug was defined as 0% cytotoxicity. Error bars indicate SD. ** P ≤0.01, and ***, P ≤0.001 compared to nano-curcumin. * indicates µg apotransferrin protein that carry equivalent molar concentration of the drug.

    Article Snippet: Nanoparticle localization assay SUPT1 cells obtained from the NIH-AIDS Reference and Reagents Program were used . (1×106 ) SUPT1 cells or stimulated PBMCs were seeded in 30 mm dishes (Corning Lifesciences) and treated with curcumin, either soluble or incorporated into nanoparticles at 1 and 10 µM concentrations and the cells were incubated at different time points (1, 2, 4 and 6 h).

    Techniques: MTT Assay, Cell Culture, Concentration Assay

    Nano-curcumin more effectively inhibits HIV-1 replication through a mechanism dependent on transferrin receptor. A C) SUPT1 cells (Panel A) or stimulated PBMCs (Panel C) were challenged for 2 h with HIV-1 93IN101 (1 mg p24/ml) in the presence of increasing concentrations (1, 2.5, 5, 10, 20 and 30 µM) of sol-curcumin, nano-curcumin, or nano-apotransferrin (10 and 50 µg). They were then incubated for a further 96 h, after which viral replication was measured by p24 antigen capture assay. *indicates µg apotransferrin protein that carry equivalent molar concentration of the drug. B D) SUP-T1 cells or stimulated PBMCs were challenged for 2 h with HIV-1 93IN101 in the presence of 2.5 or 5.0 µM concentrations of sol-curcumin, nano-curcumin or nano-curcumin in the presence of transferrin receptor antibody (100 ng/ml). After 96 h incubation, viral replication was measured by p24 antigen capture assay. In both these experiments, viral replication in the absence of drug was defined as 0% inhibition; Azidothymidine (AZT) was employed as a positive control. Error bars indicate SD. ** P ≤0.01, and ***, P ≤0.001 compared to sol-curcumin; n.s.: non-significant.

    Journal: PLoS ONE

    Article Title: Curcumin-Loaded Apotransferrin Nanoparticles Provide Efficient Cellular Uptake and Effectively Inhibit HIV-1 Replication In Vitro

    doi: 10.1371/journal.pone.0023388

    Figure Lengend Snippet: Nano-curcumin more effectively inhibits HIV-1 replication through a mechanism dependent on transferrin receptor. A C) SUPT1 cells (Panel A) or stimulated PBMCs (Panel C) were challenged for 2 h with HIV-1 93IN101 (1 mg p24/ml) in the presence of increasing concentrations (1, 2.5, 5, 10, 20 and 30 µM) of sol-curcumin, nano-curcumin, or nano-apotransferrin (10 and 50 µg). They were then incubated for a further 96 h, after which viral replication was measured by p24 antigen capture assay. *indicates µg apotransferrin protein that carry equivalent molar concentration of the drug. B D) SUP-T1 cells or stimulated PBMCs were challenged for 2 h with HIV-1 93IN101 in the presence of 2.5 or 5.0 µM concentrations of sol-curcumin, nano-curcumin or nano-curcumin in the presence of transferrin receptor antibody (100 ng/ml). After 96 h incubation, viral replication was measured by p24 antigen capture assay. In both these experiments, viral replication in the absence of drug was defined as 0% inhibition; Azidothymidine (AZT) was employed as a positive control. Error bars indicate SD. ** P ≤0.01, and ***, P ≤0.001 compared to sol-curcumin; n.s.: non-significant.

    Article Snippet: Nanoparticle localization assay SUPT1 cells obtained from the NIH-AIDS Reference and Reagents Program were used . (1×106 ) SUPT1 cells or stimulated PBMCs were seeded in 30 mm dishes (Corning Lifesciences) and treated with curcumin, either soluble or incorporated into nanoparticles at 1 and 10 µM concentrations and the cells were incubated at different time points (1, 2, 4 and 6 h).

    Techniques: Incubation, Concentration Assay, Inhibition, Positive Control

    Nanoparticle formulation increases curcumin uptake, which is inhibited by transferrin receptor blockade. A) SUP-T1 cells were incubated for 1 h with curcumin formulations as indicated, then examined by confocal microscopy. (i) Cells without curcumin; (ii) 1 µM sol-curcumin; (iii) 1 µM nano-curcumin; or (iv) 1 µM nano-curcumin in the presence of transferrin receptor antibody (100 ng/ml). Each panel contains three images: fluorescence, bright field and merged. B) SUP-T1 cells (i) or stimulated PBMCs (ii) were incubated for 1 h with curcumin formulations, after which intrinsic fluorescence of intracellular curcumin was determined quantitatively by fluorometric analysis. Cells were treated with 5 µM sol-curcumin, 5 µM nano-curcumin, or 5 µM nano-curcumin in the presence of antibodies to the transferrin receptor (TrR-Ab; 100 ng/ml). All the values are normalized to that obtained from SUPT1 cells. Error bars indicate standard deviation (SD). ***, P ≤0.001 compared to sol-curcumin; n.s.: non-significant.

    Journal: PLoS ONE

    Article Title: Curcumin-Loaded Apotransferrin Nanoparticles Provide Efficient Cellular Uptake and Effectively Inhibit HIV-1 Replication In Vitro

    doi: 10.1371/journal.pone.0023388

    Figure Lengend Snippet: Nanoparticle formulation increases curcumin uptake, which is inhibited by transferrin receptor blockade. A) SUP-T1 cells were incubated for 1 h with curcumin formulations as indicated, then examined by confocal microscopy. (i) Cells without curcumin; (ii) 1 µM sol-curcumin; (iii) 1 µM nano-curcumin; or (iv) 1 µM nano-curcumin in the presence of transferrin receptor antibody (100 ng/ml). Each panel contains three images: fluorescence, bright field and merged. B) SUP-T1 cells (i) or stimulated PBMCs (ii) were incubated for 1 h with curcumin formulations, after which intrinsic fluorescence of intracellular curcumin was determined quantitatively by fluorometric analysis. Cells were treated with 5 µM sol-curcumin, 5 µM nano-curcumin, or 5 µM nano-curcumin in the presence of antibodies to the transferrin receptor (TrR-Ab; 100 ng/ml). All the values are normalized to that obtained from SUPT1 cells. Error bars indicate standard deviation (SD). ***, P ≤0.001 compared to sol-curcumin; n.s.: non-significant.

    Article Snippet: Nanoparticle localization assay SUPT1 cells obtained from the NIH-AIDS Reference and Reagents Program were used . (1×106 ) SUPT1 cells or stimulated PBMCs were seeded in 30 mm dishes (Corning Lifesciences) and treated with curcumin, either soluble or incorporated into nanoparticles at 1 and 10 µM concentrations and the cells were incubated at different time points (1, 2, 4 and 6 h).

    Techniques: Incubation, Confocal Microscopy, Fluorescence, Standard Deviation

    Pre-transplant CMV-specific T cell responses in the patients with CMV episode. CMV-specific T cell responses were measured by stimulating PBMCs with OLPs pools of (A) pp65, (B) IE-1 in ELISPOT or (D) combination of immunodominant peptides of CMV in QuantiFERON-CMV. The combined result of (A) and (B) is represented in (C) pp65 or IE-1. In (A-C), pre-transplant CMV-specific T cell responses of the patients with post-transplant CMV episode were likely to be higher than the patients without post-transplant CMV episode. In (D), however, IFN-γ concentration was tended to lower in the patients with CMV infection. Each result was obtained by 2 repeated experiments. CMV, cytomegalovirus; PBMC, peripheral blood mononuclear cell; OLP, overlapping peptide; pp65, phosphoprotein 65; IE-1, immediate-early 1; ELISPOT, enzyme-linked immunospot; IFN, interferon.

    Journal: Immune Network

    Article Title: Comparison of the Commercial QuantiFERON-CMV and Overlapping Peptide-based ELISPOT Assays for Predicting CMV Infection in Kidney Transplant Recipients

    doi: 10.4110/in.2017.17.5.317

    Figure Lengend Snippet: Pre-transplant CMV-specific T cell responses in the patients with CMV episode. CMV-specific T cell responses were measured by stimulating PBMCs with OLPs pools of (A) pp65, (B) IE-1 in ELISPOT or (D) combination of immunodominant peptides of CMV in QuantiFERON-CMV. The combined result of (A) and (B) is represented in (C) pp65 or IE-1. In (A-C), pre-transplant CMV-specific T cell responses of the patients with post-transplant CMV episode were likely to be higher than the patients without post-transplant CMV episode. In (D), however, IFN-γ concentration was tended to lower in the patients with CMV infection. Each result was obtained by 2 repeated experiments. CMV, cytomegalovirus; PBMC, peripheral blood mononuclear cell; OLP, overlapping peptide; pp65, phosphoprotein 65; IE-1, immediate-early 1; ELISPOT, enzyme-linked immunospot; IFN, interferon.

    Article Snippet: OLPs-based ELISPOT Peripheral blood was collected from each patient before transplantation, and peripheral blood mononuclear cells (PBMCs) were isolated using Lymphocyte Separation Medium (Corning, New York, NY, USA).

    Techniques: Enzyme-linked Immunospot, Concentration Assay, Infection

    Inference and effect of barcode multiplets in single-cell ATAC-seq data. (a) Default t-SNE depiction of public scATAC-seq PBMC 5k dataset. Colors represent cluster annotations from the automated CellRanger output. (b) Quantification of barcodes affected by barcode multiplets for the same dataset (identified by bap). (c) Depiction of two multiplets each composed of 9 oligonucleotide barcodes. Barcodes in each multiplet share a long common subsequence, denoted in black. (d) Visualization of two barcode multiplets from (c) in t-SNE coordinates. (e) Visualization of all implicated barcode multiplets from this dataset. The zoomed panel shows a small group of cells affected by five multiplets, indicated by color. (f) Empirical distribution of the mean restricted longest common subsequence (rLCS) per multiplet. A cutoff of 6 was used to determine either of the two classes of barcode multiplets. (g) Percent difference of the mean log2 fragments between pairs of barcodes within a multiplet. The reported p-value is from a two-sided Kolmogorov–Smirnov test. Boxplots: center line, median; box limits, first and third quartiles; whiskers, 1.5x interquartile range. (h) Overall rates of barcode multiplets from additional scATAC-seq data comparing v1.0 and v1.1 (NextGEM) chip designs.

    Journal: bioRxiv

    Article Title: Inference and effects of barcode multiplets in droplet-based single-cell assays

    doi: 10.1101/824003

    Figure Lengend Snippet: Inference and effect of barcode multiplets in single-cell ATAC-seq data. (a) Default t-SNE depiction of public scATAC-seq PBMC 5k dataset. Colors represent cluster annotations from the automated CellRanger output. (b) Quantification of barcodes affected by barcode multiplets for the same dataset (identified by bap). (c) Depiction of two multiplets each composed of 9 oligonucleotide barcodes. Barcodes in each multiplet share a long common subsequence, denoted in black. (d) Visualization of two barcode multiplets from (c) in t-SNE coordinates. (e) Visualization of all implicated barcode multiplets from this dataset. The zoomed panel shows a small group of cells affected by five multiplets, indicated by color. (f) Empirical distribution of the mean restricted longest common subsequence (rLCS) per multiplet. A cutoff of 6 was used to determine either of the two classes of barcode multiplets. (g) Percent difference of the mean log2 fragments between pairs of barcodes within a multiplet. The reported p-value is from a two-sided Kolmogorov–Smirnov test. Boxplots: center line, median; box limits, first and third quartiles; whiskers, 1.5x interquartile range. (h) Overall rates of barcode multiplets from additional scATAC-seq data comparing v1.0 and v1.1 (NextGEM) chip designs.

    Article Snippet: Profiling PBMCs using 10x scATAC-seq For 10x scATAC-seq experiments with PBMCs (PB003F, Allcells), frozen cells were quickly thawed in a 37°C water bath for about 30s and transferred to a 15 mL tube.

    Techniques: Chromatin Immunoprecipitation

    Supporting information for Figure 3 . (a) Quantification of barcodes affected by barcode multiplets for the PBMC dataset generated with this work (“This Study”). (b) Percentage of barcode multiplets identified for different numbers of input barcodes (see Methods ). (c) Visualization of seven additional barcode multiplets from the Public dataset. (d) Proportion of bead pairs occurring in the same chromatin accessibility-defined Louvain cluster compared to a permuted background. Error bars represent standard error of mean over 100 permutations per dataset. (e) Downsampling analysis of the dataset generated in this work (“This Study”). Barcode multiplets were examined at downsampled intervals from 10%-90% by units of 10%. The highlighted sample represents 40% downsampling and corresponds to a median 10,000 fragments detected per barcode. At all downsampled thresholds, we detected 0 pairs that were not present in the 100% sample. (f) Distribution of the restricted longest common subsequence (rLCS) for 1,000,000 randomly-sampled barcode pairs in the 10x barcode universe. A threshold at 6 is drawn for use in other analyses.

    Journal: bioRxiv

    Article Title: Inference and effects of barcode multiplets in droplet-based single-cell assays

    doi: 10.1101/824003

    Figure Lengend Snippet: Supporting information for Figure 3 . (a) Quantification of barcodes affected by barcode multiplets for the PBMC dataset generated with this work (“This Study”). (b) Percentage of barcode multiplets identified for different numbers of input barcodes (see Methods ). (c) Visualization of seven additional barcode multiplets from the Public dataset. (d) Proportion of bead pairs occurring in the same chromatin accessibility-defined Louvain cluster compared to a permuted background. Error bars represent standard error of mean over 100 permutations per dataset. (e) Downsampling analysis of the dataset generated in this work (“This Study”). Barcode multiplets were examined at downsampled intervals from 10%-90% by units of 10%. The highlighted sample represents 40% downsampling and corresponds to a median 10,000 fragments detected per barcode. At all downsampled thresholds, we detected 0 pairs that were not present in the 100% sample. (f) Distribution of the restricted longest common subsequence (rLCS) for 1,000,000 randomly-sampled barcode pairs in the 10x barcode universe. A threshold at 6 is drawn for use in other analyses.

    Article Snippet: Profiling PBMCs using 10x scATAC-seq For 10x scATAC-seq experiments with PBMCs (PB003F, Allcells), frozen cells were quickly thawed in a 37°C water bath for about 30s and transferred to a 15 mL tube.

    Techniques: Generated

    ( a ) p24 viral antigen levels 10 days post-infection in the supernatant of HIV-1 infected PBMCs treated with TIZ (10 μg/mL) or RM-4848 (10 μg/mL). Results obtained with PBMC of 20 different healthy donors are shown. Median values and statistical significance is presented. ( b ) Inhibition (%) of viral replication in PBMC of 20 different healthy donors infected in vitro with HIV-1 in the presence of TIZ- (10 μg/mL) or RM-4848- (10 μg/mL). Median values and statistical significance is presented. ( c ) p24 viral antigen levels in PBMCs infected with HIV-1 in the presence of RM-4848 throughout infection (RM-4848)(10 μg/mL) or in medium alone (no treatment). In addition to these two situations, results of 3 other experimental conditions are shown: 1) PBMCs treated with RM-4848 before being infected (pre-infection); 2) RM-4848 present during the 24 hours infection period (during infection) and then washed away; 3) RM-4848 added 24 hours after infection (post infection). Mean values ± standard errors are shown.

    Journal: Scientific Reports

    Article Title: Thiazolides Elicit Anti-Viral Innate Immunity and Reduce HIV Replication

    doi: 10.1038/srep27148

    Figure Lengend Snippet: ( a ) p24 viral antigen levels 10 days post-infection in the supernatant of HIV-1 infected PBMCs treated with TIZ (10 μg/mL) or RM-4848 (10 μg/mL). Results obtained with PBMC of 20 different healthy donors are shown. Median values and statistical significance is presented. ( b ) Inhibition (%) of viral replication in PBMC of 20 different healthy donors infected in vitro with HIV-1 in the presence of TIZ- (10 μg/mL) or RM-4848- (10 μg/mL). Median values and statistical significance is presented. ( c ) p24 viral antigen levels in PBMCs infected with HIV-1 in the presence of RM-4848 throughout infection (RM-4848)(10 μg/mL) or in medium alone (no treatment). In addition to these two situations, results of 3 other experimental conditions are shown: 1) PBMCs treated with RM-4848 before being infected (pre-infection); 2) RM-4848 present during the 24 hours infection period (during infection) and then washed away; 3) RM-4848 added 24 hours after infection (post infection). Mean values ± standard errors are shown.

    Article Snippet: PBMC isolation and HIV-1 infection Whole blood was collected from 20 healthy volunteers by venipuncture in Vacutainer tubes containing EDTA (Becton Dickinson, Franklin Lakes, NJ), and PBMCs were separated on lymphocyte separation medium (Organon Teknica, Malvern, PA).

    Techniques: Infection, Inhibition, In Vitro

    Cytokines (IL-2, IFNγ) and chemokines (MCP-1, MIP-1α, MIP-1β, RANTES) production (pg/ml) by PBMCs of healthy donors. PBMCs were either uninfected (white bars) or HIV-1-infected (grey bars) and were cultured in medium alone (no drug), or in medium containing either TIZ- (10 μg/mL) ( a ) or RM-4848- (10 μg/mL) ( b ). Mean values 7 days post-infection ± standard errors and statistical differences are shown.

    Journal: Scientific Reports

    Article Title: Thiazolides Elicit Anti-Viral Innate Immunity and Reduce HIV Replication

    doi: 10.1038/srep27148

    Figure Lengend Snippet: Cytokines (IL-2, IFNγ) and chemokines (MCP-1, MIP-1α, MIP-1β, RANTES) production (pg/ml) by PBMCs of healthy donors. PBMCs were either uninfected (white bars) or HIV-1-infected (grey bars) and were cultured in medium alone (no drug), or in medium containing either TIZ- (10 μg/mL) ( a ) or RM-4848- (10 μg/mL) ( b ). Mean values 7 days post-infection ± standard errors and statistical differences are shown.

    Article Snippet: PBMC isolation and HIV-1 infection Whole blood was collected from 20 healthy volunteers by venipuncture in Vacutainer tubes containing EDTA (Becton Dickinson, Franklin Lakes, NJ), and PBMCs were separated on lymphocyte separation medium (Organon Teknica, Malvern, PA).

    Techniques: Infection, Cell Culture

    mRNA expression of 84 genes that are part of the HIV-1 life cycle and known to directly obstacle HIV infectivity has been assessed by real-time quantitative RT-PCR, calculated relative to five housekeeping genes and shown as fold-change expression from the untreated sample. Gene expression (nfold) is shown as a color scale from green to red (−2 to +9) (MEV multiple experiment viewer software). Results were obtained in HIV1-infected PBMCs at day 7 post infection, cultured in medium containing TIZ (10 μg/mL) or RM-4848 (10 μg/mL). Only targets showing > 2-fold modulation are considered significant and are shown in tab.

    Journal: Scientific Reports

    Article Title: Thiazolides Elicit Anti-Viral Innate Immunity and Reduce HIV Replication

    doi: 10.1038/srep27148

    Figure Lengend Snippet: mRNA expression of 84 genes that are part of the HIV-1 life cycle and known to directly obstacle HIV infectivity has been assessed by real-time quantitative RT-PCR, calculated relative to five housekeeping genes and shown as fold-change expression from the untreated sample. Gene expression (nfold) is shown as a color scale from green to red (−2 to +9) (MEV multiple experiment viewer software). Results were obtained in HIV1-infected PBMCs at day 7 post infection, cultured in medium containing TIZ (10 μg/mL) or RM-4848 (10 μg/mL). Only targets showing > 2-fold modulation are considered significant and are shown in tab.

    Article Snippet: PBMC isolation and HIV-1 infection Whole blood was collected from 20 healthy volunteers by venipuncture in Vacutainer tubes containing EDTA (Becton Dickinson, Franklin Lakes, NJ), and PBMCs were separated on lymphocyte separation medium (Organon Teknica, Malvern, PA).

    Techniques: Expressing, Infection, Quantitative RT-PCR, Software, Cell Culture

    mRNA expression of genes involved in cholesterol metabolism and efflux in PBMCs of healthy donors. ( a ) Uninfected PBMCs cultured in medium containing TIZ (10 μg/mL); ( b ) Uninfected PBMCs cultured in medium containing RM-4848 (10 μg/mL); ( c ) HIV-1–infected PBMCs (day 7 post infection) cultured in medium containing TIZ (10 μg/mL); ( d ) HIV-1–infected PBMCs (day 7 post infection) cultured in medium containing RM-4848 (10 μg/mL). Gene expression genes is assessed by quantitative RT-PCR and shown as fold-change expression from the untreated sample. Only targets showing > 2-fold modulation are considered significant. Mean values ± S.D. and p values are indicated.

    Journal: Scientific Reports

    Article Title: Thiazolides Elicit Anti-Viral Innate Immunity and Reduce HIV Replication

    doi: 10.1038/srep27148

    Figure Lengend Snippet: mRNA expression of genes involved in cholesterol metabolism and efflux in PBMCs of healthy donors. ( a ) Uninfected PBMCs cultured in medium containing TIZ (10 μg/mL); ( b ) Uninfected PBMCs cultured in medium containing RM-4848 (10 μg/mL); ( c ) HIV-1–infected PBMCs (day 7 post infection) cultured in medium containing TIZ (10 μg/mL); ( d ) HIV-1–infected PBMCs (day 7 post infection) cultured in medium containing RM-4848 (10 μg/mL). Gene expression genes is assessed by quantitative RT-PCR and shown as fold-change expression from the untreated sample. Only targets showing > 2-fold modulation are considered significant. Mean values ± S.D. and p values are indicated.

    Article Snippet: PBMC isolation and HIV-1 infection Whole blood was collected from 20 healthy volunteers by venipuncture in Vacutainer tubes containing EDTA (Becton Dickinson, Franklin Lakes, NJ), and PBMCs were separated on lymphocyte separation medium (Organon Teknica, Malvern, PA).

    Techniques: Expressing, Cell Culture, Infection, Quantitative RT-PCR

    p24 viral antigen concentration in HIV-1 - infected PBMCs cultured in medium alone (no drug) or in the presence of TO-901317 (1 μM), an LXR agonist. Results were obtained with PBMCs of 20 different healthy donors 10 days post-infection. Mean values ± standard errors and statistical significance are shown.

    Journal: Scientific Reports

    Article Title: Thiazolides Elicit Anti-Viral Innate Immunity and Reduce HIV Replication

    doi: 10.1038/srep27148

    Figure Lengend Snippet: p24 viral antigen concentration in HIV-1 - infected PBMCs cultured in medium alone (no drug) or in the presence of TO-901317 (1 μM), an LXR agonist. Results were obtained with PBMCs of 20 different healthy donors 10 days post-infection. Mean values ± standard errors and statistical significance are shown.

    Article Snippet: PBMC isolation and HIV-1 infection Whole blood was collected from 20 healthy volunteers by venipuncture in Vacutainer tubes containing EDTA (Becton Dickinson, Franklin Lakes, NJ), and PBMCs were separated on lymphocyte separation medium (Organon Teknica, Malvern, PA).

    Techniques: Concentration Assay, Infection, Cell Culture

    Rituximab ( RTX ) leads to further phenotypical changes in CD56 dim natural killer ( NK ) cells. Freshly isolated peripheral blood mononuclear cells (PBMCs) were incubated with (+) or without ( w/o ) RTX overnight. The next day PBMCs were stained with a mixture of antibodies (anti-CD3 PE-Dazzle, anti-CD56 Bv421 and anti-CD16 FITC and CD69/CD137 PE) and analyzed by flow cytometry. NK cells were identified as CD3-CD56+ cells in the lymphocyte gate and fluorescence intensity of PE on CD56 dim NK cells was measured. a CD69 PE. Histogram shows one representative donor; gray incubation + RTX, black w/o RTX. Left diagram mean CD69 PE fluorescence intensity in samples from 13 donors. Statistical significance was determined with the Wilcoxon signed rank test (p = 0.0002). Right diagram Δ % in CD56 dim NK was determined by (CD69-positive CD56 dim NK cells after incubation + RTX) - (CD69-positive CD56 dim NK cells after incubation w/o RTX); bar median. b CD137(41BB)-PE; same donor, layout and abbreviations as in a ; n = 11 donors; p = 0.001

    Journal: Arthritis Research & Therapy

    Article Title: Rituximab induces phenotypical and functional changes of NK cells in a non-malignant experimental setting

    doi: 10.1186/s13075-016-1101-3

    Figure Lengend Snippet: Rituximab ( RTX ) leads to further phenotypical changes in CD56 dim natural killer ( NK ) cells. Freshly isolated peripheral blood mononuclear cells (PBMCs) were incubated with (+) or without ( w/o ) RTX overnight. The next day PBMCs were stained with a mixture of antibodies (anti-CD3 PE-Dazzle, anti-CD56 Bv421 and anti-CD16 FITC and CD69/CD137 PE) and analyzed by flow cytometry. NK cells were identified as CD3-CD56+ cells in the lymphocyte gate and fluorescence intensity of PE on CD56 dim NK cells was measured. a CD69 PE. Histogram shows one representative donor; gray incubation + RTX, black w/o RTX. Left diagram mean CD69 PE fluorescence intensity in samples from 13 donors. Statistical significance was determined with the Wilcoxon signed rank test (p = 0.0002). Right diagram Δ % in CD56 dim NK was determined by (CD69-positive CD56 dim NK cells after incubation + RTX) - (CD69-positive CD56 dim NK cells after incubation w/o RTX); bar median. b CD137(41BB)-PE; same donor, layout and abbreviations as in a ; n = 11 donors; p = 0.001

    Article Snippet: Flow cytometry PBMCs were stained for 20–30 minutes on ice with a cocktail of monoclonal antibodies, then washed and directly analyzed on a four-laser flow cytometer (LSR Fortessa, BD Biosciences, San Jose, CA, USA).

    Techniques: Isolation, Incubation, Staining, Flow Cytometry, Cytometry, Fluorescence

    Addition of rituximab to peripheral blood mononuclear cells ( PMBCs ) leads to B cell depletion in the absence of serum. a PBMCs were isolated by density gradient centrifugation, left untreated overnight, and cultured over the second night in medium supplemented with heat-inactivated fetal calf serum without any therapeutic antibody ( w/o , left panel ), with 10 μg/ml anti-CD20 antibody rituximab ( RTX ) ( middle ) or with 10 μg/ml anti-TNF alpha antibody infliximab ( right panel ). After that, PBMCs were stained with a mixture of lineage antibodies (anti-CD3 PE-Dazzle, anti-CD56 Bv421, anti-CD19 FITC) and analyzed by flow cytometry. B cells were identified as CD3-CD19+ cells in the lymphocyte gate. b PBMCs were treated as described ( a ). After washing PBMCs were re-diluted in Annexin-V (AxV) buffer supplemented with PE-labeled Annexin-V and analyzed by flow cytometry. B cells were identified as CD3-CD19+ cells in the lymphocyte gate. Upper row culture in medium without RTX ( w/o RTX ). Lower row culture in medium with RTX ( + RTX ). AxV+ B cells apoptotic B cells with positive staining for Annexin-V PE. a , b B cell numbers were reduced after incubation with RTX in 14/14 independent experiments, each performed with PBMCs from different healthy donors. In 10/14 experiments RTX-induced B cell depletion was complete as shown ( a ); in 4/14 B cell depletion was incomplete as shown ( b ). Infliximab was used as a negative control in 2/14 experiments. Increased binding of Annexin-V was identified in three donors with incomplete B cell depletion. c Left graph B cell percentages before and after culture overnight with rituximab (statistically significant, Wilcoxon signed rank test, p = 0.001). Right graph , whiskers 10–90th percentile. PBMCs with incomplete B cell depletion after incubation with RTX overnight ( incomplete ) had significantly lower ratios of natural killer cells to B cells ( NK/B ) than PBMCSs with complete B cell depletion ( complete ,

    Journal: Arthritis Research & Therapy

    Article Title: Rituximab induces phenotypical and functional changes of NK cells in a non-malignant experimental setting

    doi: 10.1186/s13075-016-1101-3

    Figure Lengend Snippet: Addition of rituximab to peripheral blood mononuclear cells ( PMBCs ) leads to B cell depletion in the absence of serum. a PBMCs were isolated by density gradient centrifugation, left untreated overnight, and cultured over the second night in medium supplemented with heat-inactivated fetal calf serum without any therapeutic antibody ( w/o , left panel ), with 10 μg/ml anti-CD20 antibody rituximab ( RTX ) ( middle ) or with 10 μg/ml anti-TNF alpha antibody infliximab ( right panel ). After that, PBMCs were stained with a mixture of lineage antibodies (anti-CD3 PE-Dazzle, anti-CD56 Bv421, anti-CD19 FITC) and analyzed by flow cytometry. B cells were identified as CD3-CD19+ cells in the lymphocyte gate. b PBMCs were treated as described ( a ). After washing PBMCs were re-diluted in Annexin-V (AxV) buffer supplemented with PE-labeled Annexin-V and analyzed by flow cytometry. B cells were identified as CD3-CD19+ cells in the lymphocyte gate. Upper row culture in medium without RTX ( w/o RTX ). Lower row culture in medium with RTX ( + RTX ). AxV+ B cells apoptotic B cells with positive staining for Annexin-V PE. a , b B cell numbers were reduced after incubation with RTX in 14/14 independent experiments, each performed with PBMCs from different healthy donors. In 10/14 experiments RTX-induced B cell depletion was complete as shown ( a ); in 4/14 B cell depletion was incomplete as shown ( b ). Infliximab was used as a negative control in 2/14 experiments. Increased binding of Annexin-V was identified in three donors with incomplete B cell depletion. c Left graph B cell percentages before and after culture overnight with rituximab (statistically significant, Wilcoxon signed rank test, p = 0.001). Right graph , whiskers 10–90th percentile. PBMCs with incomplete B cell depletion after incubation with RTX overnight ( incomplete ) had significantly lower ratios of natural killer cells to B cells ( NK/B ) than PBMCSs with complete B cell depletion ( complete ,

    Article Snippet: Flow cytometry PBMCs were stained for 20–30 minutes on ice with a cocktail of monoclonal antibodies, then washed and directly analyzed on a four-laser flow cytometer (LSR Fortessa, BD Biosciences, San Jose, CA, USA).

    Techniques: Isolation, Gradient Centrifugation, Cell Culture, Staining, Flow Cytometry, Cytometry, Labeling, Incubation, Negative Control, Binding Assay

    Rituximab ( RTX ) leads to natural killer ( NK ) cell degranulation and downregulation of CD16 in peripheral blood mononuclear cells (PBMCs). PBMCs were isolated and cultured as described in Fig. 1 . Anti-CD107a PE-Cy5 was added at the same time point as the therapeutic antibodies (starting point of the degranulation assay). The next day PBMCs were stained with a mixture of antibodies (anti-CD3 PE, anti-CD56 Bv421 and anti-CD16 FITC) and analyzed by flow cytometry. NK cells were identified as CD3-CD56+ cells in the lymphocyte gate. a CD107a expression on CD56 dim NK cells after stimulation without ( w/o ) therapeutic antibody, with 10 μg/ml RTX (+ RTX ), with 10 μg/ml infliximab and with 10 μg/ml intravenous immunoglobulin ( IvIg ) (from left to right ). Shown is one representative donor. b Summary of the increased CD107a expression on total NK cells after treatment with RTX in comparison to infliximab. Dots linked by a line belong to the same donor. Additive effect on degranulation is defined by (CD107a pos. NK cells after culture with therapeutic antibody) -(CD107a pos. NK cells after culture without therapeutic antibody). c CD16 expression on CD107a-positive NK cells. Shown is one representative donor. a - c CD107a expression together with CD16 expression has been investigated in healthy individuals ( n = 6 (+/− RTX), n = 3 (+/− infliximab) and n = 2 (+/− IvIg)). d The percentage of CD16 bright cells among CD56 dim NK cells before and after stimulation with RTX overnight was investigated in 16 healthy donors. Statistical significance was determined with the Wilcoxon signed rank test ( p = 0.0005). FSC forward scatter

    Journal: Arthritis Research & Therapy

    Article Title: Rituximab induces phenotypical and functional changes of NK cells in a non-malignant experimental setting

    doi: 10.1186/s13075-016-1101-3

    Figure Lengend Snippet: Rituximab ( RTX ) leads to natural killer ( NK ) cell degranulation and downregulation of CD16 in peripheral blood mononuclear cells (PBMCs). PBMCs were isolated and cultured as described in Fig. 1 . Anti-CD107a PE-Cy5 was added at the same time point as the therapeutic antibodies (starting point of the degranulation assay). The next day PBMCs were stained with a mixture of antibodies (anti-CD3 PE, anti-CD56 Bv421 and anti-CD16 FITC) and analyzed by flow cytometry. NK cells were identified as CD3-CD56+ cells in the lymphocyte gate. a CD107a expression on CD56 dim NK cells after stimulation without ( w/o ) therapeutic antibody, with 10 μg/ml RTX (+ RTX ), with 10 μg/ml infliximab and with 10 μg/ml intravenous immunoglobulin ( IvIg ) (from left to right ). Shown is one representative donor. b Summary of the increased CD107a expression on total NK cells after treatment with RTX in comparison to infliximab. Dots linked by a line belong to the same donor. Additive effect on degranulation is defined by (CD107a pos. NK cells after culture with therapeutic antibody) -(CD107a pos. NK cells after culture without therapeutic antibody). c CD16 expression on CD107a-positive NK cells. Shown is one representative donor. a - c CD107a expression together with CD16 expression has been investigated in healthy individuals ( n = 6 (+/− RTX), n = 3 (+/− infliximab) and n = 2 (+/− IvIg)). d The percentage of CD16 bright cells among CD56 dim NK cells before and after stimulation with RTX overnight was investigated in 16 healthy donors. Statistical significance was determined with the Wilcoxon signed rank test ( p = 0.0005). FSC forward scatter

    Article Snippet: Flow cytometry PBMCs were stained for 20–30 minutes on ice with a cocktail of monoclonal antibodies, then washed and directly analyzed on a four-laser flow cytometer (LSR Fortessa, BD Biosciences, San Jose, CA, USA).

    Techniques: Isolation, Cell Culture, Degranulation Assay, Staining, Flow Cytometry, Cytometry, Expressing

    The extent of rituximab-induced B cell depletion correlates with the size of natural killer ( NK ) cell proportions. NK cells were isolated from one fraction of freshly isolated peripheral blood mononuclear cells (PBMCs) and left untreated overnight. Another fraction of freshly isolated PBMCs was left untreated overnight and NK cells were depleted the next day using anti-CD56 and anti-CD16 monoclonal antibodies and magnetic beads. NK-cell-depleted PBMCs were resubstituted with isolated NK cells in three different proportions. The samples were incubated with ( gray bars ) or without ( black bars ) 10 μg/ml rituximab overnight. PBMCs were stained and analyzed by flow cytometry as described in Fig. 1 . a - c Three independent experiments. First bars show NK cell depletion without re-substitution. Second to fourth bars show NK cell depletion with resubstitution of different percentages of NK cells as indicated. a Data from the same experiment shown in Fig. 3 (cultured without human serum); the complete experiment with all negative controls including proof of successful NK cell depletion is shown in Additional file 1 : Figure S1(b). b , c Two further experiments performed with different donors (cultured with autologous human serum). The statistical analysis is described in “ Results ”

    Journal: Arthritis Research & Therapy

    Article Title: Rituximab induces phenotypical and functional changes of NK cells in a non-malignant experimental setting

    doi: 10.1186/s13075-016-1101-3

    Figure Lengend Snippet: The extent of rituximab-induced B cell depletion correlates with the size of natural killer ( NK ) cell proportions. NK cells were isolated from one fraction of freshly isolated peripheral blood mononuclear cells (PBMCs) and left untreated overnight. Another fraction of freshly isolated PBMCs was left untreated overnight and NK cells were depleted the next day using anti-CD56 and anti-CD16 monoclonal antibodies and magnetic beads. NK-cell-depleted PBMCs were resubstituted with isolated NK cells in three different proportions. The samples were incubated with ( gray bars ) or without ( black bars ) 10 μg/ml rituximab overnight. PBMCs were stained and analyzed by flow cytometry as described in Fig. 1 . a - c Three independent experiments. First bars show NK cell depletion without re-substitution. Second to fourth bars show NK cell depletion with resubstitution of different percentages of NK cells as indicated. a Data from the same experiment shown in Fig. 3 (cultured without human serum); the complete experiment with all negative controls including proof of successful NK cell depletion is shown in Additional file 1 : Figure S1(b). b , c Two further experiments performed with different donors (cultured with autologous human serum). The statistical analysis is described in “ Results ”

    Article Snippet: Flow cytometry PBMCs were stained for 20–30 minutes on ice with a cocktail of monoclonal antibodies, then washed and directly analyzed on a four-laser flow cytometer (LSR Fortessa, BD Biosciences, San Jose, CA, USA).

    Techniques: Isolation, Magnetic Beads, Incubation, Staining, Flow Cytometry, Cytometry, Cell Culture

    Inhibition of cytokine secretion in LPS-stimulated PBMCs by rhIL-1ra. LPS, lipopolysaccharide; PBMCs, peripheral blood mononuclear cells; rh, human recombinant; IL, interleukin; ra, receptor antagonist; low, LPS + 125 ng/ml rhIL-1ra; middle, LPS + 250 ng/ml rhIL-1ra; high, LPS + 500 ng/ml rhIL-1ra; IFN, interferon; TNF, tumor necrosis factor.

    Journal: Experimental and Therapeutic Medicine

    Article Title: Interleukin-1 receptor antagonist expression is inversely associated with outcomes of hepatitis B-related acute-on-chronic liver failure

    doi: 10.3892/etm.2017.4361

    Figure Lengend Snippet: Inhibition of cytokine secretion in LPS-stimulated PBMCs by rhIL-1ra. LPS, lipopolysaccharide; PBMCs, peripheral blood mononuclear cells; rh, human recombinant; IL, interleukin; ra, receptor antagonist; low, LPS + 125 ng/ml rhIL-1ra; middle, LPS + 250 ng/ml rhIL-1ra; high, LPS + 500 ng/ml rhIL-1ra; IFN, interferon; TNF, tumor necrosis factor.

    Article Snippet: Incubation of peripheral blood mononuclear cells (PBMCs) with IL-1ra in vitro PBMCs were isolated from the heparinized blood of 31 patients with ACLF via centrifugation at 800 × g at 20°C for 20 min using Ficoll Lymphoprep (Axis-Shield Diagnostics Ltd., Dundee, UK).

    Techniques: Inhibition, Recombinant