blood mononuclear cells pbmcs Search Results


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  • 97
    Lonza peripheral blood mononuclear cells pbmcs
    Peripheral Blood Mononuclear Cells Pbmcs, supplied by Lonza, used in various techniques. Bioz Stars score: 97/100, based on 126 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ATCC primary peripheral blood mononuclear cells pbmc normal human
    Tensor factorization to map model-predicted cytokine responses. A) Measured receptor abundance for ten <t>PBMC-derived</t> subpopulations. Points and error bars show geometric mean and standard deviation respectively (N = 4). Error bars for some points are too small to display. B-C) PCA scores (B) and loadings (C) of receptor abundance. Axis label percentages indicate percent variance explained. D) Schematic representation of CP decomposition. Model predictions are arranged in a cube depending upon the time, ligand treatment, and cell type being modeled. CP decomposition then helps to visualize this space. E) Percent variance reconstructed (R2X) versus the number of components used in non-negative CP decomposition. F-I) Component values versus time (F), cell type (G-H), or ligand stimulation (I). The variation explained by each component is the product of the component’s time, ligand, and cell type factorization. Ligand components with only negligible values (
    Primary Peripheral Blood Mononuclear Cells Pbmc Normal Human, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Axis-Shield Diagnostics peripheral blood mononuclear cells pbmcs
    Lenalidomide maintenance therapy does not enhance MM-specific CD4 + T-cell responses Analysis of T-cell responses specific for the MAGE-C1 peptide SQSSPVSSFPSSTSS. ( A ) CD137 expression on CD4 + and CD8 + T cells from patient#02 at the pre <t>LEN</t> timepoint upon in vitro restimulation with the MAGE-C1 peptide. ( B ) <t>PBMCs</t> obtained at the pre LEN and the LEN timepoint were examined by IFN-γ ELISPOT to measure the T-cell response to the MAGE-C1 peptide. The bar graphs show the mean number of spots per million cells of four replicate wells and the error bars represent the standard deviation. A two-way ANOVA with Sidak’s multiple comparisons test was performed to evaluate statistical significant differences, ns: not significant, ** p ≤ 0.01, *** p ≤ 0.001 and **** p ≤ 0.0001.
    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
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    Becton Dickinson peripheral blood mononuclear cells pbmcs
    Measuring CTLA-4 and Fas expression by <t>CD4</t> + ,FOXP3 + T cells as well as apoptosis of CD4 + ,CD25 bright and CD25 − T cells in young and elderly humans. (A – D) <t>PBMCs</t> from young (n = 15) and elderly (n = 15) individuals were stained with Abs
    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
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    Nycomed peripheral blood mononuclear cells pbmc
    CD69 expression in response to CMV antigens and to the influenza vaccine. <t>PBMCs</t> from elderly individuals were stimulated for 18 h and CD69 expression was evaluated by flow cytometry. a Representative dot plots of the frequency of CMV- and influenza vaccine-specific <t>CD4</t> T cells with and without NKG2D expression. Percentages of positive cells in this representative experiment are expressed in dot plots. b Histograms summarize the percentage of CD69-cells positive for CMV and influenza vaccine in the CD4+ CD28 null NKG2D+ ( black bars ) and CD4+ CD28 null NKG2D- subsets ( white bars ) obtained from 11 elderly donors. Paired t test was used to compare paired frequencies
    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
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    Biochrom peripheral blood mononuclear cells pbmcs
    Single cell profiling identifies CD96 high and CD96 low subsets of Th9 cells. ( A – D ) Single cell gene expression analysis of CD44 + alloreactive T cells sorted from nonpolarized (w/o; n = 100), Th1 ( n = 67), and Th9 ( n = 113) polarized BALB/c cocultures. See Table S2 for a list of all genes measured. ( A ) Principal component analysis of gene expression in indicated Th subsets. ( B ) Nonsupervised hierarchical clustering analysis (Pearson’s correlation) comparing gene expression in Th subsets. Analysis includes all genes with detectable expression. ( C ) Box plots of most differentially expressed genes. Bonferroni-corrected ANOVA P values: Il9 ( P = 1.4 × 10 −126 ), Il2ra ( P = 1.4 × 10 −65 ), Cd83 ( P = 7.1 × 10 −64 ), Il4ra ( P = 1.2 × 10 −52 ), Gata3 ( P = 3.6 × 10 −47 ), Il1r1 ( P = 5.5 × 10 −47 ), Ccr4 ( P = 2.2 × 10 −46 ), and Cd28 ( P = 2.3 × 10 −43 ). ( D ) Nonsupervised hierarchical clustering analysis (complete linkage method) of gene expression in CD44 + T cells sorted from alloreactive BALB/c Th9 cultures. Displayed genes contributed to clustering. Frame indicates a cluster of Th9 cells characterized by high Cd96 expression. ( E ) Contour plots of surface CD96 expression in CD44 + polarized alloreactive BALB/c T cells, including fluorescence minus one (FMO) staining control (Ctrl). ( F ) Comparison of CD96 expression levels in IL-9 + and IFN-γ + human <t>CD4</t> + T cells. <t>PBMCs</t> from blood of 10 healthy individuals were stimulated ex vivo with Staphylococcus enterotoxin B (SEB) for 24 h. A one-tailed Wilcoxon matched-pairs signed rank test was applied. gMFI, geometric mean fluorescence intensity.
    Peripheral Blood Mononuclear Cells Pbmcs, supplied by Biochrom, used in various techniques. Bioz Stars score: 92/100, based on 407 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    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 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Becton Dickinson blood mononuclear cells pbmcs
    Frequency and molecular expression of pDC in IT, CHB and AHB groups. (a) AHB group had significantly higher <t>PBMC</t> percent in peripheral blood leukocytes, compared with CHB or IT group. (b) Frequency of pDC in PBMC decreased in AHB and CHB groups, compared with IT group. (c) The difference in CD86+ pDC frequency among three groups was not significant. (d) The counts of CD86 molecular expressed on surface of pDC in CHB group was higher, compared with AHB and IT groups. IT group: HBeAg-positive chronic hepatitis B virus infected patients in immune tolerance phase; CHB group: HBeAg-positive chronic hepatitis B infected patients; AHB group: Acute hepatitis B virus infected patients; PBMC: Peripheral blood mononuclear cells; pDC: <t>Plasmacytoid</t> dendritic cell; CD86: Cluster of differentiation antigen 86.
    Blood Mononuclear Cells Pbmcs, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 93/100, based on 339 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Valiant peripheral blood mononuclear cells pbmcs
    Exosomes from tumor patients with increased membrane-associated HSP72 and HSP105 induced <t>DCs</t> to promote tumor cell invasion via IL-6 in vitro . (A) After adsorption onto latex beads, HSP72 and HSP105 on EXO-PT and EXO-TT were detected by flow cytometry. RFI was calculated by dividing mean fluorescence intensity of samples staining with detection Abs by that of sample staining with ISO Abs (n = 7). (B) Human DCs were induced from <t>PBMCs</t> and stimulated with 5 μg/ml of EXO-PT or EXO-TT from breast tumor tissues for 6 h. The IL-6 production was measured by ELISA (n = 3). (C) MDA-MB-435S cells were cultured in supernatant from PBS-, EXO-PT- or EXO-TT (5 μg/ml for 6 h)-stimulated DCs with or without 30 μg/ml of anti-IL-6 or ISO mAbs for 24 h. Then, their invasive ability was measured using an in vitro invasive assay, and the cells on the bottom of the Transwell filter were imaged and quantified (Magnification: 200 ×). (D) The results of (C) were statistically analyzed (n = 5). (E, F) After adsorption onto latex beads, HSP72 and HSP105 on exosomes from sera of breast tumor patients or healthy controls were detected by flow cytometry. One representative result of 12 is shown (E). RFI was calculated and statistically analyzed (F). (B, D) The results are shown as the mean ± SEM of 3 independent experiments. (C) One representative of 3 independent experiments is shown. P values were generated by 2-tail student's t -test in A, F, and by one-way ANOVA, followed by a Tukey-Kramer multiple comparison test in B, D; *** 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
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    Millipore blood mononuclear cells pbmcs
    Exosomes from tumor patients with increased membrane-associated HSP72 and HSP105 induced <t>DCs</t> to promote tumor cell invasion via IL-6 in vitro . (A) After adsorption onto latex beads, HSP72 and HSP105 on EXO-PT and EXO-TT were detected by flow cytometry. RFI was calculated by dividing mean fluorescence intensity of samples staining with detection Abs by that of sample staining with ISO Abs (n = 7). (B) Human DCs were induced from <t>PBMCs</t> and stimulated with 5 μg/ml of EXO-PT or EXO-TT from breast tumor tissues for 6 h. The IL-6 production was measured by ELISA (n = 3). (C) MDA-MB-435S cells were cultured in supernatant from PBS-, EXO-PT- or EXO-TT (5 μg/ml for 6 h)-stimulated DCs with or without 30 μg/ml of anti-IL-6 or ISO mAbs for 24 h. Then, their invasive ability was measured using an in vitro invasive assay, and the cells on the bottom of the Transwell filter were imaged and quantified (Magnification: 200 ×). (D) The results of (C) were statistically analyzed (n = 5). (E, F) After adsorption onto latex beads, HSP72 and HSP105 on exosomes from sera of breast tumor patients or healthy controls were detected by flow cytometry. One representative result of 12 is shown (E). RFI was calculated and statistically analyzed (F). (B, D) The results are shown as the mean ± SEM of 3 independent experiments. (C) One representative of 3 independent experiments is shown. P values were generated by 2-tail student's t -test in A, F, and by one-way ANOVA, followed by a Tukey-Kramer multiple comparison test in B, D; *** p
    Blood Mononuclear Cells Pbmcs, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 524 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore peripheral blood mononuclear cells
    Exosomes from tumor patients with increased membrane-associated HSP72 and HSP105 induced <t>DCs</t> to promote tumor cell invasion via IL-6 in vitro . (A) After adsorption onto latex beads, HSP72 and HSP105 on EXO-PT and EXO-TT were detected by flow cytometry. RFI was calculated by dividing mean fluorescence intensity of samples staining with detection Abs by that of sample staining with ISO Abs (n = 7). (B) Human DCs were induced from <t>PBMCs</t> and stimulated with 5 μg/ml of EXO-PT or EXO-TT from breast tumor tissues for 6 h. The IL-6 production was measured by ELISA (n = 3). (C) MDA-MB-435S cells were cultured in supernatant from PBS-, EXO-PT- or EXO-TT (5 μg/ml for 6 h)-stimulated DCs with or without 30 μg/ml of anti-IL-6 or ISO mAbs for 24 h. Then, their invasive ability was measured using an in vitro invasive assay, and the cells on the bottom of the Transwell filter were imaged and quantified (Magnification: 200 ×). (D) The results of (C) were statistically analyzed (n = 5). (E, F) After adsorption onto latex beads, HSP72 and HSP105 on exosomes from sera of breast tumor patients or healthy controls were detected by flow cytometry. One representative result of 12 is shown (E). RFI was calculated and statistically analyzed (F). (B, D) The results are shown as the mean ± SEM of 3 independent experiments. (C) One representative of 3 independent experiments is shown. P values were generated by 2-tail student's t -test in A, F, and by one-way ANOVA, followed by a Tukey-Kramer multiple comparison test in B, D; *** p
    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
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    PAA Laboratories peripheral blood mononuclear cells pbmcs
    Exosomes from tumor patients with increased membrane-associated HSP72 and HSP105 induced <t>DCs</t> to promote tumor cell invasion via IL-6 in vitro . (A) After adsorption onto latex beads, HSP72 and HSP105 on EXO-PT and EXO-TT were detected by flow cytometry. RFI was calculated by dividing mean fluorescence intensity of samples staining with detection Abs by that of sample staining with ISO Abs (n = 7). (B) Human DCs were induced from <t>PBMCs</t> and stimulated with 5 μg/ml of EXO-PT or EXO-TT from breast tumor tissues for 6 h. The IL-6 production was measured by ELISA (n = 3). (C) MDA-MB-435S cells were cultured in supernatant from PBS-, EXO-PT- or EXO-TT (5 μg/ml for 6 h)-stimulated DCs with or without 30 μg/ml of anti-IL-6 or ISO mAbs for 24 h. Then, their invasive ability was measured using an in vitro invasive assay, and the cells on the bottom of the Transwell filter were imaged and quantified (Magnification: 200 ×). (D) The results of (C) were statistically analyzed (n = 5). (E, F) After adsorption onto latex beads, HSP72 and HSP105 on exosomes from sera of breast tumor patients or healthy controls were detected by flow cytometry. One representative result of 12 is shown (E). RFI was calculated and statistically analyzed (F). (B, D) The results are shown as the mean ± SEM of 3 independent experiments. (C) One representative of 3 independent experiments is shown. P values were generated by 2-tail student's t -test in A, F, and by one-way ANOVA, followed by a Tukey-Kramer multiple comparison test in B, D; *** p
    Peripheral Blood Mononuclear Cells Pbmcs, supplied by PAA Laboratories, used in various techniques. Bioz Stars score: 92/100, based on 71 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Corning Life Sciences peripheral blood mononuclear cells pbmcs
    Pre-transplant CMV-specific T cell responses in the patients with CMV episode. CMV-specific T cell responses were measured by stimulating <t>PBMCs</t> with <t>OLPs</t> 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.
    Peripheral Blood Mononuclear Cells Pbmcs, supplied by Corning Life Sciences, used in various techniques. Bioz Stars score: 90/100, based on 99 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Greiner Bio peripheral blood mononuclear cells pbmcs
    Pre-transplant CMV-specific T cell responses in the patients with CMV episode. CMV-specific T cell responses were measured by stimulating <t>PBMCs</t> with <t>OLPs</t> 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.
    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
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    Axis-Shield Diagnostics blood mononuclear cells pbmcs
    Expression of CD69 and PD-1 and apoptosis of <t>MAIT</t> cells after stimulation with IL-12 and IL-18. <t>PBMCs</t> (1 × 10 6 /well) were incubated for 24 hours in the presence of IL-12 (50 ng/mL) and IL-18 (50 ng/mL), and then stained with FITC-conjugated anti-CD3, FITC-conjugated annexin V, APC-conjugated anti-TCR Vα7.2, PE-conjugated anti-CD3, PE-conjugated anti-CD69, PE-conjugated anti-PD-1 and PE-Cy5-conjugated anti-CD161 monoclonal antibodies. Percentages of CD69-expressing cells ( panel A ), annexin V-positive cells ( panel C ), and PD-1-expressing cells ( panel E ) among MAIT cells were determined by flow cytometry. Data in panels B, D and F were obtained from 6 HCs. Values are expressed as the mean ± SEM. *p
    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
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    Mediatech peripheral blood mononuclear cells pbmcs
    Expression of CD69 and PD-1 and apoptosis of <t>MAIT</t> cells after stimulation with IL-12 and IL-18. <t>PBMCs</t> (1 × 10 6 /well) were incubated for 24 hours in the presence of IL-12 (50 ng/mL) and IL-18 (50 ng/mL), and then stained with FITC-conjugated anti-CD3, FITC-conjugated annexin V, APC-conjugated anti-TCR Vα7.2, PE-conjugated anti-CD3, PE-conjugated anti-CD69, PE-conjugated anti-PD-1 and PE-Cy5-conjugated anti-CD161 monoclonal antibodies. Percentages of CD69-expressing cells ( panel A ), annexin V-positive cells ( panel C ), and PD-1-expressing cells ( panel E ) among MAIT cells were determined by flow cytometry. Data in panels B, D and F were obtained from 6 HCs. Values are expressed as the mean ± SEM. *p
    Peripheral Blood Mononuclear Cells Pbmcs, supplied by Mediatech, used in various techniques. Bioz Stars score: 92/100, based on 204 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Sanquin peripheral blood mononuclear cells pbmc
    Expression of CD69 and PD-1 and apoptosis of <t>MAIT</t> cells after stimulation with IL-12 and IL-18. <t>PBMCs</t> (1 × 10 6 /well) were incubated for 24 hours in the presence of IL-12 (50 ng/mL) and IL-18 (50 ng/mL), and then stained with FITC-conjugated anti-CD3, FITC-conjugated annexin V, APC-conjugated anti-TCR Vα7.2, PE-conjugated anti-CD3, PE-conjugated anti-CD69, PE-conjugated anti-PD-1 and PE-Cy5-conjugated anti-CD161 monoclonal antibodies. Percentages of CD69-expressing cells ( panel A ), annexin V-positive cells ( panel C ), and PD-1-expressing cells ( panel E ) among MAIT cells were determined by flow cytometry. Data in panels B, D and F were obtained from 6 HCs. Values are expressed as the mean ± SEM. *p
    Peripheral Blood Mononuclear Cells Pbmc, supplied by Sanquin, used in various techniques. Bioz Stars score: 93/100, based on 55 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Axis-Shield Diagnostics human peripheral blood mononuclear cells pbmcs
    Infection with Mtb HN878 triggers increased IL-22 production through a TLR2-dependent pathway Lung cells isolated from B6, Il1r-/- and Tlr2-/- mice were infected with either Mtb H37Rv (Rv) or Mtb HN878 (HN) at MOI of 0.1 for 7 days, and the protein levels of ( a ) IL-1β and ( b ) IL-22 were assessed in culture supernatants by ELISA and luminex assay, respectively. Lung cells isolated from B6 mice were treated with cell wall extracted from Mtb H37Rv (Rv) or Mtb HN878 (HN), (10μg/ml each). Protein levels of ( c ) IL-1β and ( d ) IL-22 were assessed by ELISA and luminex assay respectively. <t>PBMCs</t> were isolated from <t>ATB</t> patients and were stimulated with Mtb H37Rv (Rv) or Mtb HN878 (HN) cell wall extracts (20 μg/ml each) and the protein levels of ( e ) IL-22 were measured by luminex assays. Error bars represent means ± s.e.m. n = 3-5 for a-d. n = 17 for human ATB patients for e. ND, not detected; UI, uninfected; UN, untreated. * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001 by ( a,b ) one-way ANOVA, ( c,d ) Student's t test ( e ) two-way ANOVA
    Human Peripheral Blood Mononuclear Cells Pbmcs, supplied by Axis-Shield Diagnostics, used in various techniques. Bioz Stars score: 93/100, based on 112 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    BioLegend veri cells cd4 low pbmc
    Infection with Mtb HN878 triggers increased IL-22 production through a TLR2-dependent pathway Lung cells isolated from B6, Il1r-/- and Tlr2-/- mice were infected with either Mtb H37Rv (Rv) or Mtb HN878 (HN) at MOI of 0.1 for 7 days, and the protein levels of ( a ) IL-1β and ( b ) IL-22 were assessed in culture supernatants by ELISA and luminex assay, respectively. Lung cells isolated from B6 mice were treated with cell wall extracted from Mtb H37Rv (Rv) or Mtb HN878 (HN), (10μg/ml each). Protein levels of ( c ) IL-1β and ( d ) IL-22 were assessed by ELISA and luminex assay respectively. <t>PBMCs</t> were isolated from <t>ATB</t> patients and were stimulated with Mtb H37Rv (Rv) or Mtb HN878 (HN) cell wall extracts (20 μg/ml each) and the protein levels of ( e ) IL-22 were measured by luminex assays. Error bars represent means ± s.e.m. n = 3-5 for a-d. n = 17 for human ATB patients for e. ND, not detected; UI, uninfected; UN, untreated. * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001 by ( a,b ) one-way ANOVA, ( c,d ) Student's t test ( e ) two-way ANOVA
    Veri Cells Cd4 Low Pbmc, supplied by BioLegend, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Fresenius Kabi peripheral blood mononuclear cells pbmcs
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    Infection with Mtb HN878 triggers increased IL-22 production through a TLR2-dependent pathway Lung cells isolated from B6, Il1r-/- and Tlr2-/- mice were infected with either Mtb H37Rv (Rv) or Mtb HN878 (HN) at MOI of 0.1 for 7 days, and the protein levels of ( a ) IL-1β and ( b ) IL-22 were assessed in culture supernatants by ELISA and luminex assay, respectively. Lung cells isolated from B6 mice were treated with cell wall extracted from Mtb H37Rv (Rv) or Mtb HN878 (HN), (10μg/ml each). Protein levels of ( c ) IL-1β and ( d ) IL-22 were assessed by ELISA and luminex assay respectively. <t>PBMCs</t> were isolated from <t>ATB</t> patients and were stimulated with Mtb H37Rv (Rv) or Mtb HN878 (HN) cell wall extracts (20 μg/ml each) and the protein levels of ( e ) IL-22 were measured by luminex assays. Error bars represent means ± s.e.m. n = 3-5 for a-d. n = 17 for human ATB patients for e. ND, not detected; UI, uninfected; UN, untreated. * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001 by ( a,b ) one-way ANOVA, ( c,d ) Student's t test ( e ) two-way ANOVA
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    Infection with Mtb HN878 triggers increased IL-22 production through a TLR2-dependent pathway Lung cells isolated from B6, Il1r-/- and Tlr2-/- mice were infected with either Mtb H37Rv (Rv) or Mtb HN878 (HN) at MOI of 0.1 for 7 days, and the protein levels of ( a ) IL-1β and ( b ) IL-22 were assessed in culture supernatants by ELISA and luminex assay, respectively. Lung cells isolated from B6 mice were treated with cell wall extracted from Mtb H37Rv (Rv) or Mtb HN878 (HN), (10μg/ml each). Protein levels of ( c ) IL-1β and ( d ) IL-22 were assessed by ELISA and luminex assay respectively. <t>PBMCs</t> were isolated from <t>ATB</t> patients and were stimulated with Mtb H37Rv (Rv) or Mtb HN878 (HN) cell wall extracts (20 μg/ml each) and the protein levels of ( e ) IL-22 were measured by luminex assays. Error bars represent means ± s.e.m. n = 3-5 for a-d. n = 17 for human ATB patients for e. ND, not detected; UI, uninfected; UN, untreated. * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001 by ( a,b ) one-way ANOVA, ( c,d ) Student's t test ( e ) two-way ANOVA
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    Mean (±95% CI ; ±min./max.) peripheral blood mononuclear cell ( <t>PBMC</t> ) nuclear concentration of <t>DNA</t> methyltransferase 3B ( DNMT 3B) following stimulation with pre‐ and post‐exercise plasma. A paired sample t ‐test was used to test significance (* P
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    Image Search Results


    Tensor factorization to map model-predicted cytokine responses. A) Measured receptor abundance for ten PBMC-derived subpopulations. Points and error bars show geometric mean and standard deviation respectively (N = 4). Error bars for some points are too small to display. B-C) PCA scores (B) and loadings (C) of receptor abundance. Axis label percentages indicate percent variance explained. D) Schematic representation of CP decomposition. Model predictions are arranged in a cube depending upon the time, ligand treatment, and cell type being modeled. CP decomposition then helps to visualize this space. E) Percent variance reconstructed (R2X) versus the number of components used in non-negative CP decomposition. F-I) Component values versus time (F), cell type (G-H), or ligand stimulation (I). The variation explained by each component is the product of the component’s time, ligand, and cell type factorization. Ligand components with only negligible values (

    Journal: bioRxiv

    Article Title: Modeling Cell-Specific Dynamics and Regulation of the Common Gamma Chain Cytokines

    doi: 10.1101/778894

    Figure Lengend Snippet: Tensor factorization to map model-predicted cytokine responses. A) Measured receptor abundance for ten PBMC-derived subpopulations. Points and error bars show geometric mean and standard deviation respectively (N = 4). Error bars for some points are too small to display. B-C) PCA scores (B) and loadings (C) of receptor abundance. Axis label percentages indicate percent variance explained. D) Schematic representation of CP decomposition. Model predictions are arranged in a cube depending upon the time, ligand treatment, and cell type being modeled. CP decomposition then helps to visualize this space. E) Percent variance reconstructed (R2X) versus the number of components used in non-negative CP decomposition. F-I) Component values versus time (F), cell type (G-H), or ligand stimulation (I). The variation explained by each component is the product of the component’s time, ligand, and cell type factorization. Ligand components with only negligible values (

    Article Snippet: Receptor abundance quantitation Cryopreserved PBMCs (ATCC, PCS-800-011, lot#81115172) were thawed to room temperature and slowly diluted with 9 mL pre-warmed RPMI-1640 medium (Gibco, 11875-093) supplemented with 10% fetal bovine serum (FBS, Seradigm, 1500-500, lot#322B15).

    Techniques: Derivative Assay, Standard Deviation

    Receptor quantification and gating of PBMC-derived immune cell types. A) Preliminary gating for single lyphocytes. B) Example staining for CD122 (red), the corresponding isotype control (blue), and unstained cells (black). C) Gating for live T helper and T regulatory cells during receptor quantification. D) Live cell NK cell gating. E) Live cell CD8+ T cell gating. F) Gating for fixed T helper and T regulatory cells during pSTAT5 quantification. G) Fixed CD8+ T cell and NK cell gating.

    Journal: bioRxiv

    Article Title: Modeling Cell-Specific Dynamics and Regulation of the Common Gamma Chain Cytokines

    doi: 10.1101/778894

    Figure Lengend Snippet: Receptor quantification and gating of PBMC-derived immune cell types. A) Preliminary gating for single lyphocytes. B) Example staining for CD122 (red), the corresponding isotype control (blue), and unstained cells (black). C) Gating for live T helper and T regulatory cells during receptor quantification. D) Live cell NK cell gating. E) Live cell CD8+ T cell gating. F) Gating for fixed T helper and T regulatory cells during pSTAT5 quantification. G) Fixed CD8+ T cell and NK cell gating.

    Article Snippet: Receptor abundance quantitation Cryopreserved PBMCs (ATCC, PCS-800-011, lot#81115172) were thawed to room temperature and slowly diluted with 9 mL pre-warmed RPMI-1640 medium (Gibco, 11875-093) supplemented with 10% fetal bovine serum (FBS, Seradigm, 1500-500, lot#322B15).

    Techniques: Derivative Assay, Staining

    Model accurately predicts cell type-specific response across a panel of PBMC-derived cell types. A) Comparison of two replicates measuring pSTAT5 response to a dose-response of IL-2/-15, time course, and panel of PBMC-derived cell types. B) Both experimentally-derived and model-predicted EC 50 s of dose response across IL-2/-15 and all 10 cell types. EC 50 s are shown for 1 hr time point. C) Pearson correlation coefficients between model prediction and experimental measurements for all 10 cell populations (full data shown in Fig. S5 ). D–I) pSTAT5 response to IL-2 (D-F) or IL-15 (G-I) dose responses in NK, CD8+, and T reg cells.

    Journal: bioRxiv

    Article Title: Modeling Cell-Specific Dynamics and Regulation of the Common Gamma Chain Cytokines

    doi: 10.1101/778894

    Figure Lengend Snippet: Model accurately predicts cell type-specific response across a panel of PBMC-derived cell types. A) Comparison of two replicates measuring pSTAT5 response to a dose-response of IL-2/-15, time course, and panel of PBMC-derived cell types. B) Both experimentally-derived and model-predicted EC 50 s of dose response across IL-2/-15 and all 10 cell types. EC 50 s are shown for 1 hr time point. C) Pearson correlation coefficients between model prediction and experimental measurements for all 10 cell populations (full data shown in Fig. S5 ). D–I) pSTAT5 response to IL-2 (D-F) or IL-15 (G-I) dose responses in NK, CD8+, and T reg cells.

    Article Snippet: Receptor abundance quantitation Cryopreserved PBMCs (ATCC, PCS-800-011, lot#81115172) were thawed to room temperature and slowly diluted with 9 mL pre-warmed RPMI-1640 medium (Gibco, 11875-093) supplemented with 10% fetal bovine serum (FBS, Seradigm, 1500-500, lot#322B15).

    Techniques: Derivative Assay

    A reaction model captures cytokine-cytokine interactions. A) Schematic of IL-4 and IL-7 receptor complexes competing for γ c and generating distinct pSTAT signals. B-C) Fitting model to experimental data. Experimental measurements are denoted by triangles. Shaded areas represent the 25-75% and 10-90% confidence intervals of model predictions. pSTAT5 and pSTAT6 were measured for IL-7 and IL-4 experiments, respectively. B) Singlecytokine pSTAT dose-response measurements for 10 min of exposure to IL-4 and IL-7. C) Percent inhibition of the second cytokine’s pSTAT response in a dual-cytokine dose-response experiment. Human PBMC-derived T cells (CD4 + TCR + CCR7 high ) were pretreated with various concentrations of one cytokine for 10 min before being stimulated with a fixed concentration (50 pg/mL IL-7 or 100 pg/mL IL-4) of the other cytokine for an additional 10 min. D) Model predictions for percent inhibition of the second cytokine’s pSTAT response in a dual-cytokine dose-response experiment with the assumption that active species are endocytosed at the same rate as inactive species (k endo,a = k endo ). E) Model predictions for percent of γ c on the cell surface when exposed to 100 pg/mL of either IL-7 or IL-4 for 100 min. F) Violin plot of K a values obtained via posterior distributions of k fwd / k rev for krev parameters corresponding to different complexes competing for the common γ c ( Fig. 1A ). G–I) Posterior distributions from fitting to data. Scaling constants C 5 and C 6 have units of # × cell −1 , k fwd has units of cell × # −1 × min −1 , and f sort is unitless

    Journal: bioRxiv

    Article Title: Modeling Cell-Specific Dynamics and Regulation of the Common Gamma Chain Cytokines

    doi: 10.1101/778894

    Figure Lengend Snippet: A reaction model captures cytokine-cytokine interactions. A) Schematic of IL-4 and IL-7 receptor complexes competing for γ c and generating distinct pSTAT signals. B-C) Fitting model to experimental data. Experimental measurements are denoted by triangles. Shaded areas represent the 25-75% and 10-90% confidence intervals of model predictions. pSTAT5 and pSTAT6 were measured for IL-7 and IL-4 experiments, respectively. B) Singlecytokine pSTAT dose-response measurements for 10 min of exposure to IL-4 and IL-7. C) Percent inhibition of the second cytokine’s pSTAT response in a dual-cytokine dose-response experiment. Human PBMC-derived T cells (CD4 + TCR + CCR7 high ) were pretreated with various concentrations of one cytokine for 10 min before being stimulated with a fixed concentration (50 pg/mL IL-7 or 100 pg/mL IL-4) of the other cytokine for an additional 10 min. D) Model predictions for percent inhibition of the second cytokine’s pSTAT response in a dual-cytokine dose-response experiment with the assumption that active species are endocytosed at the same rate as inactive species (k endo,a = k endo ). E) Model predictions for percent of γ c on the cell surface when exposed to 100 pg/mL of either IL-7 or IL-4 for 100 min. F) Violin plot of K a values obtained via posterior distributions of k fwd / k rev for krev parameters corresponding to different complexes competing for the common γ c ( Fig. 1A ). G–I) Posterior distributions from fitting to data. Scaling constants C 5 and C 6 have units of # × cell −1 , k fwd has units of cell × # −1 × min −1 , and f sort is unitless

    Article Snippet: Receptor abundance quantitation Cryopreserved PBMCs (ATCC, PCS-800-011, lot#81115172) were thawed to room temperature and slowly diluted with 9 mL pre-warmed RPMI-1640 medium (Gibco, 11875-093) supplemented with 10% fetal bovine serum (FBS, Seradigm, 1500-500, lot#322B15).

    Techniques: Inhibition, Derivative Assay, Concentration Assay

    Lenalidomide maintenance therapy does not enhance MM-specific CD4 + T-cell responses Analysis of T-cell responses specific for the MAGE-C1 peptide SQSSPVSSFPSSTSS. ( A ) CD137 expression on CD4 + and CD8 + T cells from patient#02 at the pre LEN timepoint upon in vitro restimulation with the MAGE-C1 peptide. ( B ) PBMCs obtained at the pre LEN and the LEN timepoint were examined by IFN-γ ELISPOT to measure the T-cell response to the MAGE-C1 peptide. The bar graphs show the mean number of spots per million cells of four replicate wells and the error bars represent the standard deviation. A two-way ANOVA with Sidak’s multiple comparisons test was performed to evaluate statistical significant differences, ns: not significant, ** p ≤ 0.01, *** p ≤ 0.001 and **** p ≤ 0.0001.

    Journal: Oncotarget

    Article Title: Impact of lenalidomide maintenance on the immune environment of multiple myeloma patients with low tumor burden after autologous stem cell transplantation

    doi: 10.18632/oncotarget.24944

    Figure Lengend Snippet: Lenalidomide maintenance therapy does not enhance MM-specific CD4 + T-cell responses Analysis of T-cell responses specific for the MAGE-C1 peptide SQSSPVSSFPSSTSS. ( A ) CD137 expression on CD4 + and CD8 + T cells from patient#02 at the pre LEN timepoint upon in vitro restimulation with the MAGE-C1 peptide. ( B ) PBMCs obtained at the pre LEN and the LEN timepoint were examined by IFN-γ ELISPOT to measure the T-cell response to the MAGE-C1 peptide. The bar graphs show the mean number of spots per million cells of four replicate wells and the error bars represent the standard deviation. A two-way ANOVA with Sidak’s multiple comparisons test was performed to evaluate statistical significant differences, ns: not significant, ** p ≤ 0.01, *** p ≤ 0.001 and **** p ≤ 0.0001.

    Article Snippet: Peripheral blood mononuclear cells (PBMCs) were isolated from leukapheresis (pre-LEN timepoint) and whole blood (LEN timepoint) samples through lymphoprep gradient-centrifugation (Axis-Shield) following the manufacturer’s instructions before being cryopreserved.

    Techniques: Expressing, In Vitro, Enzyme-linked Immunospot, Standard Deviation

    Lenalidomide maintenance therapy results in an improved CD8 + T cell functionality, which is hampered by non-CD8 + T cells present in the periphery ( A ) PBMCs obtained at the pre LEN and the LEN timepoint were examined by IFN-γ ELISPOT to measure the CD8 + T-cell response to viral recall epitopes derived for CMV, EBV and influenza (CEF) ( n = 6). The number of spot forming units (SFU) as well as the activity in each well was determined. Bar graphs show the mean number of spots per million PBMCs or the mean activity of four replicate wells and the error bars show the standard deviation. Spot numbers and activity measured upon culture in the absence of peptide were subtracted. ( B – E ) CD8 + T cells purified from PBMC samples obtained at the pre LEN and LEN timepoint were examined by IFN-γ/IL-2/TNF-α FLUOROSPOT to characterize the CD8 + T-cell response to the CEF peptide pool. (B–C) Bar graphs show the mean number of spots per million CD8 + T cells of five replicate wells and the error bars show the standard deviation. Spot numbers and activity measured upon culture in the absence of peptide were subtracted. ( D ) Raw data showing three-color FLUOROSPOT results. One representative well is shown for each patient and timepoint. Cytokine-specific spots are shown in green (IFN-γ), red (TNF-α), blue (IL-2) or overlays of these colors. ( E ) The pie charts indicate the portion of mono- and polyfunctional CEF-specific CD8 + T cells. A two-way ANOVA with Sidak’s multiple comparisons test was performed to detect differences between the pre LEN and the LEN timepoint, ns: not significant, * p

    Journal: Oncotarget

    Article Title: Impact of lenalidomide maintenance on the immune environment of multiple myeloma patients with low tumor burden after autologous stem cell transplantation

    doi: 10.18632/oncotarget.24944

    Figure Lengend Snippet: Lenalidomide maintenance therapy results in an improved CD8 + T cell functionality, which is hampered by non-CD8 + T cells present in the periphery ( A ) PBMCs obtained at the pre LEN and the LEN timepoint were examined by IFN-γ ELISPOT to measure the CD8 + T-cell response to viral recall epitopes derived for CMV, EBV and influenza (CEF) ( n = 6). The number of spot forming units (SFU) as well as the activity in each well was determined. Bar graphs show the mean number of spots per million PBMCs or the mean activity of four replicate wells and the error bars show the standard deviation. Spot numbers and activity measured upon culture in the absence of peptide were subtracted. ( B – E ) CD8 + T cells purified from PBMC samples obtained at the pre LEN and LEN timepoint were examined by IFN-γ/IL-2/TNF-α FLUOROSPOT to characterize the CD8 + T-cell response to the CEF peptide pool. (B–C) Bar graphs show the mean number of spots per million CD8 + T cells of five replicate wells and the error bars show the standard deviation. Spot numbers and activity measured upon culture in the absence of peptide were subtracted. ( D ) Raw data showing three-color FLUOROSPOT results. One representative well is shown for each patient and timepoint. Cytokine-specific spots are shown in green (IFN-γ), red (TNF-α), blue (IL-2) or overlays of these colors. ( E ) The pie charts indicate the portion of mono- and polyfunctional CEF-specific CD8 + T cells. A two-way ANOVA with Sidak’s multiple comparisons test was performed to detect differences between the pre LEN and the LEN timepoint, ns: not significant, * p

    Article Snippet: Peripheral blood mononuclear cells (PBMCs) were isolated from leukapheresis (pre-LEN timepoint) and whole blood (LEN timepoint) samples through lymphoprep gradient-centrifugation (Axis-Shield) following the manufacturer’s instructions before being cryopreserved.

    Techniques: Enzyme-linked Immunospot, Derivative Assay, Activity Assay, Standard Deviation, Purification

    Measuring CTLA-4 and Fas expression by CD4 + ,FOXP3 + T cells as well as apoptosis of CD4 + ,CD25 bright and CD25 − T cells in young and elderly humans. (A – D) PBMCs from young (n = 15) and elderly (n = 15) individuals were stained with Abs

    Journal: Mechanisms of ageing and development

    Article Title: Aging and human CD4+ regulatory T cells

    doi: 10.1016/j.mad.2009.06.003

    Figure Lengend Snippet: Measuring CTLA-4 and Fas expression by CD4 + ,FOXP3 + T cells as well as apoptosis of CD4 + ,CD25 bright and CD25 − T cells in young and elderly humans. (A – D) PBMCs from young (n = 15) and elderly (n = 15) individuals were stained with Abs

    Article Snippet: Peripheral blood mononuclear cells (PBMCs) were stained with mouse anti-human CD4 and CD25 antibodies (Abs) (BD Pharmingen, San Jose, CA) followed by washing, permeabilization with permeabilizing buffer and staining with mouse anti-FOXP3 Abs (clone PCH101, eBioscience, San Diego, CA).

    Techniques: Expressing, Staining

    Correlation of CD25, FOXP3 and IL-7Rα expression by CD4 + T cells in young and elderly humans. Purified peripheral blood mononuclear cells (PBMCs) from young (n = 10) and elderly (n = 10) individuals were stained with Abs to CD4 and CD25, followed

    Journal: Mechanisms of ageing and development

    Article Title: Aging and human CD4+ regulatory T cells

    doi: 10.1016/j.mad.2009.06.003

    Figure Lengend Snippet: Correlation of CD25, FOXP3 and IL-7Rα expression by CD4 + T cells in young and elderly humans. Purified peripheral blood mononuclear cells (PBMCs) from young (n = 10) and elderly (n = 10) individuals were stained with Abs to CD4 and CD25, followed

    Article Snippet: Peripheral blood mononuclear cells (PBMCs) were stained with mouse anti-human CD4 and CD25 antibodies (Abs) (BD Pharmingen, San Jose, CA) followed by washing, permeabilization with permeabilizing buffer and staining with mouse anti-FOXP3 Abs (clone PCH101, eBioscience, San Diego, CA).

    Techniques: Expressing, Purification, Staining

    Comparing the frequency of CD4 + ,FOXP3 + T cells and their naïve (CD45RA + ) and memory (CD45RA − ) phenotypes between young and elderly humans. PBMCs from young (n = 15) and elderly (n = 15) individuals were stained with Abs to CD4, CD45RA

    Journal: Mechanisms of ageing and development

    Article Title: Aging and human CD4+ regulatory T cells

    doi: 10.1016/j.mad.2009.06.003

    Figure Lengend Snippet: Comparing the frequency of CD4 + ,FOXP3 + T cells and their naïve (CD45RA + ) and memory (CD45RA − ) phenotypes between young and elderly humans. PBMCs from young (n = 15) and elderly (n = 15) individuals were stained with Abs to CD4, CD45RA

    Article Snippet: Peripheral blood mononuclear cells (PBMCs) were stained with mouse anti-human CD4 and CD25 antibodies (Abs) (BD Pharmingen, San Jose, CA) followed by washing, permeabilization with permeabilizing buffer and staining with mouse anti-FOXP3 Abs (clone PCH101, eBioscience, San Diego, CA).

    Techniques: Staining

    Comparing the capacity of CD4 + ,CD25 bright T cells in suppressing the proliferation and cytokine production of CD4 + ,CD25 − T cells between young and elderly humans. PBMCs from young and elderly individuals were stained with Abs to CD4 and CD25 and

    Journal: Mechanisms of ageing and development

    Article Title: Aging and human CD4+ regulatory T cells

    doi: 10.1016/j.mad.2009.06.003

    Figure Lengend Snippet: Comparing the capacity of CD4 + ,CD25 bright T cells in suppressing the proliferation and cytokine production of CD4 + ,CD25 − T cells between young and elderly humans. PBMCs from young and elderly individuals were stained with Abs to CD4 and CD25 and

    Article Snippet: Peripheral blood mononuclear cells (PBMCs) were stained with mouse anti-human CD4 and CD25 antibodies (Abs) (BD Pharmingen, San Jose, CA) followed by washing, permeabilization with permeabilizing buffer and staining with mouse anti-FOXP3 Abs (clone PCH101, eBioscience, San Diego, CA).

    Techniques: Staining

    Chemokine receptor expression on CD4 + ,FOXP3 + T cells in young and elderly humans. PBMCs from young (n = 15) and elderly (n = 15) individuals were stained with Abs to CD4, CCR7, CCR4, CCR5, CXCR3 or isotype Abs, followed by permeabilization and staining

    Journal: Mechanisms of ageing and development

    Article Title: Aging and human CD4+ regulatory T cells

    doi: 10.1016/j.mad.2009.06.003

    Figure Lengend Snippet: Chemokine receptor expression on CD4 + ,FOXP3 + T cells in young and elderly humans. PBMCs from young (n = 15) and elderly (n = 15) individuals were stained with Abs to CD4, CCR7, CCR4, CCR5, CXCR3 or isotype Abs, followed by permeabilization and staining

    Article Snippet: Peripheral blood mononuclear cells (PBMCs) were stained with mouse anti-human CD4 and CD25 antibodies (Abs) (BD Pharmingen, San Jose, CA) followed by washing, permeabilization with permeabilizing buffer and staining with mouse anti-FOXP3 Abs (clone PCH101, eBioscience, San Diego, CA).

    Techniques: Expressing, Staining

    CD69 expression in response to CMV antigens and to the influenza vaccine. PBMCs from elderly individuals were stimulated for 18 h and CD69 expression was evaluated by flow cytometry. a Representative dot plots of the frequency of CMV- and influenza vaccine-specific CD4 T cells with and without NKG2D expression. Percentages of positive cells in this representative experiment are expressed in dot plots. b Histograms summarize the percentage of CD69-cells positive for CMV and influenza vaccine in the CD4+ CD28 null NKG2D+ ( black bars ) and CD4+ CD28 null NKG2D- subsets ( white bars ) obtained from 11 elderly donors. Paired t test was used to compare paired frequencies

    Journal: Age

    Article Title: NKG2D expression in CD4+ T lymphocytes as a marker of senescence in the aged immune system

    doi: 10.1007/s11357-010-9200-6

    Figure Lengend Snippet: CD69 expression in response to CMV antigens and to the influenza vaccine. PBMCs from elderly individuals were stimulated for 18 h and CD69 expression was evaluated by flow cytometry. a Representative dot plots of the frequency of CMV- and influenza vaccine-specific CD4 T cells with and without NKG2D expression. Percentages of positive cells in this representative experiment are expressed in dot plots. b Histograms summarize the percentage of CD69-cells positive for CMV and influenza vaccine in the CD4+ CD28 null NKG2D+ ( black bars ) and CD4+ CD28 null NKG2D- subsets ( white bars ) obtained from 11 elderly donors. Paired t test was used to compare paired frequencies

    Article Snippet: To analyze the differentiation status of CD4+ T cells, peripheral blood mononuclear cells (PBMC) were isolated by centrifugation on Ficoll–Hypaque gradients (Lymphoprep; Nycomed, Oslo, Norway).

    Techniques: Expressing, Flow Cytometry, Cytometry

    Differentiated status of CD4+ subsets defined by IL-2/IFN-γ production and TREC content. a PBMCs were stimulated for 6 h with soluble anti-CD3 (1 μg/mL). The responder cells were analyzed for intracellular IL-2-APC staining in the indicated populations. b Cells were treated as described previously and IFN-γ expression was analyzed by intracellular staining. Percentages of positive cells in the indicated populations in this representative experiment are expressed on the histogram-plots. Histograms summarize the percentage of IFN-γ-positive cells in CD4+ CD28 null and in CD4+ CD28+ lymphocytes NKG2D− and NKG2D + (mean ± SEM) from the six elderly donors tested. A paired t test was used to compare paired means. c Dose-response curves of the IFN-γ production by CD4 subsets defined by CD28 and NKG2D expression in response to anti-CD3. The cells were cultured for 6 h in medium alone or with increasing concentrations of anti-CD3 (1–1,000 ng/mL). A fluorescence analysis was carried out as previously described. One representative experiment of three is shown. d Quantification of TREC copy number. CD28+, CD28 null NKG2D − , and CD28 null NKG2D+ populations were isolated by sorting (magnetic bead separation) and the TREC copy number was determined by real-time PCR. Experiments were conducted in duplicate and bars represented results from three aging donors (mean ± SEM)

    Journal: Age

    Article Title: NKG2D expression in CD4+ T lymphocytes as a marker of senescence in the aged immune system

    doi: 10.1007/s11357-010-9200-6

    Figure Lengend Snippet: Differentiated status of CD4+ subsets defined by IL-2/IFN-γ production and TREC content. a PBMCs were stimulated for 6 h with soluble anti-CD3 (1 μg/mL). The responder cells were analyzed for intracellular IL-2-APC staining in the indicated populations. b Cells were treated as described previously and IFN-γ expression was analyzed by intracellular staining. Percentages of positive cells in the indicated populations in this representative experiment are expressed on the histogram-plots. Histograms summarize the percentage of IFN-γ-positive cells in CD4+ CD28 null and in CD4+ CD28+ lymphocytes NKG2D− and NKG2D + (mean ± SEM) from the six elderly donors tested. A paired t test was used to compare paired means. c Dose-response curves of the IFN-γ production by CD4 subsets defined by CD28 and NKG2D expression in response to anti-CD3. The cells were cultured for 6 h in medium alone or with increasing concentrations of anti-CD3 (1–1,000 ng/mL). A fluorescence analysis was carried out as previously described. One representative experiment of three is shown. d Quantification of TREC copy number. CD28+, CD28 null NKG2D − , and CD28 null NKG2D+ populations were isolated by sorting (magnetic bead separation) and the TREC copy number was determined by real-time PCR. Experiments were conducted in duplicate and bars represented results from three aging donors (mean ± SEM)

    Article Snippet: To analyze the differentiation status of CD4+ T cells, peripheral blood mononuclear cells (PBMC) were isolated by centrifugation on Ficoll–Hypaque gradients (Lymphoprep; Nycomed, Oslo, Norway).

    Techniques: Staining, Expressing, Cell Culture, Fluorescence, Isolation, Real-time Polymerase Chain Reaction

    Single cell profiling identifies CD96 high and CD96 low subsets of Th9 cells. ( A – D ) Single cell gene expression analysis of CD44 + alloreactive T cells sorted from nonpolarized (w/o; n = 100), Th1 ( n = 67), and Th9 ( n = 113) polarized BALB/c cocultures. See Table S2 for a list of all genes measured. ( A ) Principal component analysis of gene expression in indicated Th subsets. ( B ) Nonsupervised hierarchical clustering analysis (Pearson’s correlation) comparing gene expression in Th subsets. Analysis includes all genes with detectable expression. ( C ) Box plots of most differentially expressed genes. Bonferroni-corrected ANOVA P values: Il9 ( P = 1.4 × 10 −126 ), Il2ra ( P = 1.4 × 10 −65 ), Cd83 ( P = 7.1 × 10 −64 ), Il4ra ( P = 1.2 × 10 −52 ), Gata3 ( P = 3.6 × 10 −47 ), Il1r1 ( P = 5.5 × 10 −47 ), Ccr4 ( P = 2.2 × 10 −46 ), and Cd28 ( P = 2.3 × 10 −43 ). ( D ) Nonsupervised hierarchical clustering analysis (complete linkage method) of gene expression in CD44 + T cells sorted from alloreactive BALB/c Th9 cultures. Displayed genes contributed to clustering. Frame indicates a cluster of Th9 cells characterized by high Cd96 expression. ( E ) Contour plots of surface CD96 expression in CD44 + polarized alloreactive BALB/c T cells, including fluorescence minus one (FMO) staining control (Ctrl). ( F ) Comparison of CD96 expression levels in IL-9 + and IFN-γ + human CD4 + T cells. PBMCs from blood of 10 healthy individuals were stimulated ex vivo with Staphylococcus enterotoxin B (SEB) for 24 h. A one-tailed Wilcoxon matched-pairs signed rank test was applied. gMFI, geometric mean fluorescence intensity.

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: CD96 expression determines the inflammatory potential of IL-9–producing Th9 cells

    doi: 10.1073/pnas.1708329115

    Figure Lengend Snippet: Single cell profiling identifies CD96 high and CD96 low subsets of Th9 cells. ( A – D ) Single cell gene expression analysis of CD44 + alloreactive T cells sorted from nonpolarized (w/o; n = 100), Th1 ( n = 67), and Th9 ( n = 113) polarized BALB/c cocultures. See Table S2 for a list of all genes measured. ( A ) Principal component analysis of gene expression in indicated Th subsets. ( B ) Nonsupervised hierarchical clustering analysis (Pearson’s correlation) comparing gene expression in Th subsets. Analysis includes all genes with detectable expression. ( C ) Box plots of most differentially expressed genes. Bonferroni-corrected ANOVA P values: Il9 ( P = 1.4 × 10 −126 ), Il2ra ( P = 1.4 × 10 −65 ), Cd83 ( P = 7.1 × 10 −64 ), Il4ra ( P = 1.2 × 10 −52 ), Gata3 ( P = 3.6 × 10 −47 ), Il1r1 ( P = 5.5 × 10 −47 ), Ccr4 ( P = 2.2 × 10 −46 ), and Cd28 ( P = 2.3 × 10 −43 ). ( D ) Nonsupervised hierarchical clustering analysis (complete linkage method) of gene expression in CD44 + T cells sorted from alloreactive BALB/c Th9 cultures. Displayed genes contributed to clustering. Frame indicates a cluster of Th9 cells characterized by high Cd96 expression. ( E ) Contour plots of surface CD96 expression in CD44 + polarized alloreactive BALB/c T cells, including fluorescence minus one (FMO) staining control (Ctrl). ( F ) Comparison of CD96 expression levels in IL-9 + and IFN-γ + human CD4 + T cells. PBMCs from blood of 10 healthy individuals were stimulated ex vivo with Staphylococcus enterotoxin B (SEB) for 24 h. A one-tailed Wilcoxon matched-pairs signed rank test was applied. gMFI, geometric mean fluorescence intensity.

    Article Snippet: For quantification of intracellular cytokines in human CD4+ T cells, peripheral blood mononuclear cells (PBMCs) were isolated by Ficoll (Biochrom) gradient centrifugation and restimulated with 1 µg/mL staphylococcal enterotoxin B (SEB) (Sigma-Aldrich) for 24 h at 37 °C.

    Techniques: Expressing, Fluorescence, Staining, Ex Vivo, One-tailed Test

    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

    Frequency and molecular expression of pDC in IT, CHB and AHB groups. (a) AHB group had significantly higher PBMC percent in peripheral blood leukocytes, compared with CHB or IT group. (b) Frequency of pDC in PBMC decreased in AHB and CHB groups, compared with IT group. (c) The difference in CD86+ pDC frequency among three groups was not significant. (d) The counts of CD86 molecular expressed on surface of pDC in CHB group was higher, compared with AHB and IT groups. IT group: HBeAg-positive chronic hepatitis B virus infected patients in immune tolerance phase; CHB group: HBeAg-positive chronic hepatitis B infected patients; AHB group: Acute hepatitis B virus infected patients; PBMC: Peripheral blood mononuclear cells; pDC: Plasmacytoid dendritic cell; CD86: Cluster of differentiation antigen 86.

    Journal: Chinese Medical Journal

    Article Title: Plasmacytoid Dendritic Cell Function and Cytokine Network Profiles in Patients with Acute or Chronic Hepatitis B Virus Infection

    doi: 10.4103/0366-6999.221275

    Figure Lengend Snippet: Frequency and molecular expression of pDC in IT, CHB and AHB groups. (a) AHB group had significantly higher PBMC percent in peripheral blood leukocytes, compared with CHB or IT group. (b) Frequency of pDC in PBMC decreased in AHB and CHB groups, compared with IT group. (c) The difference in CD86+ pDC frequency among three groups was not significant. (d) The counts of CD86 molecular expressed on surface of pDC in CHB group was higher, compared with AHB and IT groups. IT group: HBeAg-positive chronic hepatitis B virus infected patients in immune tolerance phase; CHB group: HBeAg-positive chronic hepatitis B infected patients; AHB group: Acute hepatitis B virus infected patients; PBMC: Peripheral blood mononuclear cells; pDC: Plasmacytoid dendritic cell; CD86: Cluster of differentiation antigen 86.

    Article Snippet: Frequency and molecular expression of plasmacytoid dendritic cell In this study, the peripheral blood mononuclear cells (PBMCs) percentage of peripheral blood leukocytes, frequency of pDC in PBMC, frequency of cluster of differentiation antigen 86 (CD86) + pDC, and the counts of CD86 molecular expressed on surface of pDC were measured by four-color flow cytometry (FACS Caliburflow Cytometer; Becton-Dickinson, USA); and the steps to measure the expression of pDCs were as follows: the 100 μl of whole peripheral blood samples were incubated with monoclonal antibodies (mAbs) of Lin1-fluorescein isothiocyanate, human leukocyte antigen DR (HLA-DR)-peridinin chlorophyll protein, CD123-human antigen presenting cells, and CD86 - phycoerythrin (PE; all provided by BD Biosciences, Cowley, UK) inappropriate tubes at room temperature in the dark for 20 min, then added 2 ml of fluorescence activating cell sorter (FACS) Lysing solution and incubated at room temperature in the dark for 5 min, centrifuged at 300 ×g for 5 min, aspirated the supernatant, then added 2 ml of phosphate buffer saline (PBS) and vortex gently, centrifuged at 300 ×g for 5 min and aspirated the supernatant again, vortex gently and re-suspended with 200 μl PBS; finally analyzed using the FACS flow cytometer. pDCs were identified as mononuclear cells that the Lin1-(CD3-CD14-CD16-CD19-CD20-)/HLA-DR+/CD123+, the activation/maturation state of pDC was assessed by the co-stimulatory molecules CD86 in Lin1-CD123+ HLA-DR+cells.

    Techniques: Expressing, Infection

    Frequency and molecular expression of pDC in HI and HBV infected groups. (a) The difference in percentages of PBMC in peripheral blood leukocytes between two groups was not significant. Compared with HI group, frequency of pDC in PBMC was significantly lower (b), but CD86+ pDC frequency and counts of CD86 molecular expressed on surface of pDC were higher in HBV infected group (c and d). HI: Healthy individual; HBV: Hepatitis B virus; PBMC: Peripheral blood mononuclear cells; pDC: Plasmacytoid dendritic cell; CD86: Cluster of differentiation antigen 86.

    Journal: Chinese Medical Journal

    Article Title: Plasmacytoid Dendritic Cell Function and Cytokine Network Profiles in Patients with Acute or Chronic Hepatitis B Virus Infection

    doi: 10.4103/0366-6999.221275

    Figure Lengend Snippet: Frequency and molecular expression of pDC in HI and HBV infected groups. (a) The difference in percentages of PBMC in peripheral blood leukocytes between two groups was not significant. Compared with HI group, frequency of pDC in PBMC was significantly lower (b), but CD86+ pDC frequency and counts of CD86 molecular expressed on surface of pDC were higher in HBV infected group (c and d). HI: Healthy individual; HBV: Hepatitis B virus; PBMC: Peripheral blood mononuclear cells; pDC: Plasmacytoid dendritic cell; CD86: Cluster of differentiation antigen 86.

    Article Snippet: Frequency and molecular expression of plasmacytoid dendritic cell In this study, the peripheral blood mononuclear cells (PBMCs) percentage of peripheral blood leukocytes, frequency of pDC in PBMC, frequency of cluster of differentiation antigen 86 (CD86) + pDC, and the counts of CD86 molecular expressed on surface of pDC were measured by four-color flow cytometry (FACS Caliburflow Cytometer; Becton-Dickinson, USA); and the steps to measure the expression of pDCs were as follows: the 100 μl of whole peripheral blood samples were incubated with monoclonal antibodies (mAbs) of Lin1-fluorescein isothiocyanate, human leukocyte antigen DR (HLA-DR)-peridinin chlorophyll protein, CD123-human antigen presenting cells, and CD86 - phycoerythrin (PE; all provided by BD Biosciences, Cowley, UK) inappropriate tubes at room temperature in the dark for 20 min, then added 2 ml of fluorescence activating cell sorter (FACS) Lysing solution and incubated at room temperature in the dark for 5 min, centrifuged at 300 ×g for 5 min, aspirated the supernatant, then added 2 ml of phosphate buffer saline (PBS) and vortex gently, centrifuged at 300 ×g for 5 min and aspirated the supernatant again, vortex gently and re-suspended with 200 μl PBS; finally analyzed using the FACS flow cytometer. pDCs were identified as mononuclear cells that the Lin1-(CD3-CD14-CD16-CD19-CD20-)/HLA-DR+/CD123+, the activation/maturation state of pDC was assessed by the co-stimulatory molecules CD86 in Lin1-CD123+ HLA-DR+cells.

    Techniques: Expressing, Infection

    Exosomes from tumor patients with increased membrane-associated HSP72 and HSP105 induced DCs to promote tumor cell invasion via IL-6 in vitro . (A) After adsorption onto latex beads, HSP72 and HSP105 on EXO-PT and EXO-TT were detected by flow cytometry. RFI was calculated by dividing mean fluorescence intensity of samples staining with detection Abs by that of sample staining with ISO Abs (n = 7). (B) Human DCs were induced from PBMCs and stimulated with 5 μg/ml of EXO-PT or EXO-TT from breast tumor tissues for 6 h. The IL-6 production was measured by ELISA (n = 3). (C) MDA-MB-435S cells were cultured in supernatant from PBS-, EXO-PT- or EXO-TT (5 μg/ml for 6 h)-stimulated DCs with or without 30 μg/ml of anti-IL-6 or ISO mAbs for 24 h. Then, their invasive ability was measured using an in vitro invasive assay, and the cells on the bottom of the Transwell filter were imaged and quantified (Magnification: 200 ×). (D) The results of (C) were statistically analyzed (n = 5). (E, F) After adsorption onto latex beads, HSP72 and HSP105 on exosomes from sera of breast tumor patients or healthy controls were detected by flow cytometry. One representative result of 12 is shown (E). RFI was calculated and statistically analyzed (F). (B, D) The results are shown as the mean ± SEM of 3 independent experiments. (C) One representative of 3 independent experiments is shown. P values were generated by 2-tail student's t -test in A, F, and by one-way ANOVA, followed by a Tukey-Kramer multiple comparison test in B, D; *** p

    Journal: Oncoimmunology

    Article Title: Tumor-derived exosomes educate dendritic cells to promote tumor metastasis via HSP72/HSP105-TLR2/TLR4 pathway

    doi: 10.1080/2162402X.2017.1362527

    Figure Lengend Snippet: Exosomes from tumor patients with increased membrane-associated HSP72 and HSP105 induced DCs to promote tumor cell invasion via IL-6 in vitro . (A) After adsorption onto latex beads, HSP72 and HSP105 on EXO-PT and EXO-TT were detected by flow cytometry. RFI was calculated by dividing mean fluorescence intensity of samples staining with detection Abs by that of sample staining with ISO Abs (n = 7). (B) Human DCs were induced from PBMCs and stimulated with 5 μg/ml of EXO-PT or EXO-TT from breast tumor tissues for 6 h. The IL-6 production was measured by ELISA (n = 3). (C) MDA-MB-435S cells were cultured in supernatant from PBS-, EXO-PT- or EXO-TT (5 μg/ml for 6 h)-stimulated DCs with or without 30 μg/ml of anti-IL-6 or ISO mAbs for 24 h. Then, their invasive ability was measured using an in vitro invasive assay, and the cells on the bottom of the Transwell filter were imaged and quantified (Magnification: 200 ×). (D) The results of (C) were statistically analyzed (n = 5). (E, F) After adsorption onto latex beads, HSP72 and HSP105 on exosomes from sera of breast tumor patients or healthy controls were detected by flow cytometry. One representative result of 12 is shown (E). RFI was calculated and statistically analyzed (F). (B, D) The results are shown as the mean ± SEM of 3 independent experiments. (C) One representative of 3 independent experiments is shown. P values were generated by 2-tail student's t -test in A, F, and by one-way ANOVA, followed by a Tukey-Kramer multiple comparison test in B, D; *** p

    Article Snippet: To generate human DCs, peripheral blood mononuclear cells (PBMCs) from healthy volunteers were isolated by density centrifugation of heparinized blood on LSM® (MP Biomedicals), resuspended in culture medium and allowed to adhere in 6-well plates.

    Techniques: In Vitro, Adsorption, Flow Cytometry, Cytometry, Fluorescence, Staining, Enzyme-linked Immunosorbent Assay, Multiple Displacement Amplification, Cell Culture, Generated

    DCs with enhanced IL-6-secretion capacity were found in tumor-bearing mice and patients. (A, B) Mice were subcutaneously inoculated with 5 × 10 5 B16-F10 cells, and mice without tumor inoculation were used as controls. Then, DCs from splenocytes were isolated 1 week, 2 weeks or 3 weeks later and stimulated with PBS, 5 μg/ml B16-F10-EXO or 100 ng/ml of LPS for 6 h. IL-6 production by DCs was detected by ELISA (n = 3) (A). DCs from splenocytes were isolated 3 weeks later and cultured in vitro for 6 h. Culture supernatant was collected and B16-F10 cells were cultured in this supernatant for 24 h. Then, the invasive ability of B16-F10 cells was measured (n = 5). Control indicates B16-F10 cells without any treatment (B). SN-DC PBS or SN-DC tumor denotes supernatant from DCs of healthy control mice or tumor mice, respectively. (C) Mice were intravenously injected with 100 μl of PBS or 10 μg/100 μl of B16-F10-EXO. Then, DCs from splenocytes were isolated 12 or 24 h later and cultured in vitro for 6 h. IL-6 production by DCs was detected by ELISA (n = 3). (D) Human DCs were isolated from PBMCs from healthy volunteers (n = 27) or breast tumor patients (n = 18) and cultured in vitro for 6 h with or without 100 ng/ml LPS. IL-6 production by DCs was detected by ELISA. The results are shown as the mean ± SEM of 3 independent experiments or of all detected samples. P values were generated by a 2-tail student's t -test.

    Journal: Oncoimmunology

    Article Title: Tumor-derived exosomes educate dendritic cells to promote tumor metastasis via HSP72/HSP105-TLR2/TLR4 pathway

    doi: 10.1080/2162402X.2017.1362527

    Figure Lengend Snippet: DCs with enhanced IL-6-secretion capacity were found in tumor-bearing mice and patients. (A, B) Mice were subcutaneously inoculated with 5 × 10 5 B16-F10 cells, and mice without tumor inoculation were used as controls. Then, DCs from splenocytes were isolated 1 week, 2 weeks or 3 weeks later and stimulated with PBS, 5 μg/ml B16-F10-EXO or 100 ng/ml of LPS for 6 h. IL-6 production by DCs was detected by ELISA (n = 3) (A). DCs from splenocytes were isolated 3 weeks later and cultured in vitro for 6 h. Culture supernatant was collected and B16-F10 cells were cultured in this supernatant for 24 h. Then, the invasive ability of B16-F10 cells was measured (n = 5). Control indicates B16-F10 cells without any treatment (B). SN-DC PBS or SN-DC tumor denotes supernatant from DCs of healthy control mice or tumor mice, respectively. (C) Mice were intravenously injected with 100 μl of PBS or 10 μg/100 μl of B16-F10-EXO. Then, DCs from splenocytes were isolated 12 or 24 h later and cultured in vitro for 6 h. IL-6 production by DCs was detected by ELISA (n = 3). (D) Human DCs were isolated from PBMCs from healthy volunteers (n = 27) or breast tumor patients (n = 18) and cultured in vitro for 6 h with or without 100 ng/ml LPS. IL-6 production by DCs was detected by ELISA. The results are shown as the mean ± SEM of 3 independent experiments or of all detected samples. P values were generated by a 2-tail student's t -test.

    Article Snippet: To generate human DCs, peripheral blood mononuclear cells (PBMCs) from healthy volunteers were isolated by density centrifugation of heparinized blood on LSM® (MP Biomedicals), resuspended in culture medium and allowed to adhere in 6-well plates.

    Techniques: Mouse Assay, Isolation, Enzyme-linked Immunosorbent Assay, Cell Culture, In Vitro, Injection, Generated

    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

    Expression of CD69 and PD-1 and apoptosis of MAIT cells after stimulation with IL-12 and IL-18. PBMCs (1 × 10 6 /well) were incubated for 24 hours in the presence of IL-12 (50 ng/mL) and IL-18 (50 ng/mL), and then stained with FITC-conjugated anti-CD3, FITC-conjugated annexin V, APC-conjugated anti-TCR Vα7.2, PE-conjugated anti-CD3, PE-conjugated anti-CD69, PE-conjugated anti-PD-1 and PE-Cy5-conjugated anti-CD161 monoclonal antibodies. Percentages of CD69-expressing cells ( panel A ), annexin V-positive cells ( panel C ), and PD-1-expressing cells ( panel E ) among MAIT cells were determined by flow cytometry. Data in panels B, D and F were obtained from 6 HCs. Values are expressed as the mean ± SEM. *p

    Journal: PLoS Neglected Tropical Diseases

    Article Title: Activation, Impaired Tumor Necrosis Factor-α Production, and Deficiency of Circulating Mucosal-Associated Invariant T Cells in Patients with Scrub Typhus

    doi: 10.1371/journal.pntd.0004832

    Figure Lengend Snippet: Expression of CD69 and PD-1 and apoptosis of MAIT cells after stimulation with IL-12 and IL-18. PBMCs (1 × 10 6 /well) were incubated for 24 hours in the presence of IL-12 (50 ng/mL) and IL-18 (50 ng/mL), and then stained with FITC-conjugated anti-CD3, FITC-conjugated annexin V, APC-conjugated anti-TCR Vα7.2, PE-conjugated anti-CD3, PE-conjugated anti-CD69, PE-conjugated anti-PD-1 and PE-Cy5-conjugated anti-CD161 monoclonal antibodies. Percentages of CD69-expressing cells ( panel A ), annexin V-positive cells ( panel C ), and PD-1-expressing cells ( panel E ) among MAIT cells were determined by flow cytometry. Data in panels B, D and F were obtained from 6 HCs. Values are expressed as the mean ± SEM. *p

    Article Snippet: Isolation of peripheral blood mononuclear cells (PBMCs) and identification of MAIT cells Peripheral venous blood samples were collected in heparin-containing tubes, and PBMCs were isolated by density-gradient centrifugation using Lymphoprep (Axis-Shield PoC AS, Oslo, Norway).

    Techniques: Expressing, Incubation, Staining, Flow Cytometry, Cytometry

    Expression of IFN-γ, IL-17 and TNF-α in MAIT cells from scrub typhus patients. Freshly isolated PBMCs (1 × 10 6 /well) were incubated for 4 hours in the presence of PMA (100 ng/ml) and IM (1 μM). Panel A: Representative IFN-γ, IL-17 and TNF-α expression in the MAIT cell population were determined by intracellular flow cytometry after stimulation with PMA and IM. Data in panel B were obtained from 14 HCs and 23 patients with scrub typhus. Symbols represent individual subjects and horizontal lines are median values. *p

    Journal: PLoS Neglected Tropical Diseases

    Article Title: Activation, Impaired Tumor Necrosis Factor-α Production, and Deficiency of Circulating Mucosal-Associated Invariant T Cells in Patients with Scrub Typhus

    doi: 10.1371/journal.pntd.0004832

    Figure Lengend Snippet: Expression of IFN-γ, IL-17 and TNF-α in MAIT cells from scrub typhus patients. Freshly isolated PBMCs (1 × 10 6 /well) were incubated for 4 hours in the presence of PMA (100 ng/ml) and IM (1 μM). Panel A: Representative IFN-γ, IL-17 and TNF-α expression in the MAIT cell population were determined by intracellular flow cytometry after stimulation with PMA and IM. Data in panel B were obtained from 14 HCs and 23 patients with scrub typhus. Symbols represent individual subjects and horizontal lines are median values. *p

    Article Snippet: Isolation of peripheral blood mononuclear cells (PBMCs) and identification of MAIT cells Peripheral venous blood samples were collected in heparin-containing tubes, and PBMCs were isolated by density-gradient centrifugation using Lymphoprep (Axis-Shield PoC AS, Oslo, Norway).

    Techniques: Expressing, Isolation, Incubation, Flow Cytometry, Cytometry

    Expression of CD69 and PD-1 and apoptosis of MAIT cells from scrub typhus patients. Freshly isolated PBMCs were stained with FITC-conjugated anti-CD3, FITC-conjugated annexin V, APC-conjugated anti-TCR Vα7.2, PE-conjugated anti-CD3, PE-conjugated anti-CD69, PE-conjugated anti-PD-1 and PE-Cy5-conjugated anti-CD161 monoclonal antibodies, and then analyzed by flow cytometry. Percentages of CD69-expressing cells ( panel A ), annexin V-positive cells ( panel C ) and PD-1-expressing cells ( panel E ) among MAIT cells were determined by flow cytometry. Data in panels B, D and F were obtained from 14 HCs and 17 patients with scrub typhus. Symbols represent individual subjects and horizontal lines are median values. *p

    Journal: PLoS Neglected Tropical Diseases

    Article Title: Activation, Impaired Tumor Necrosis Factor-α Production, and Deficiency of Circulating Mucosal-Associated Invariant T Cells in Patients with Scrub Typhus

    doi: 10.1371/journal.pntd.0004832

    Figure Lengend Snippet: Expression of CD69 and PD-1 and apoptosis of MAIT cells from scrub typhus patients. Freshly isolated PBMCs were stained with FITC-conjugated anti-CD3, FITC-conjugated annexin V, APC-conjugated anti-TCR Vα7.2, PE-conjugated anti-CD3, PE-conjugated anti-CD69, PE-conjugated anti-PD-1 and PE-Cy5-conjugated anti-CD161 monoclonal antibodies, and then analyzed by flow cytometry. Percentages of CD69-expressing cells ( panel A ), annexin V-positive cells ( panel C ) and PD-1-expressing cells ( panel E ) among MAIT cells were determined by flow cytometry. Data in panels B, D and F were obtained from 14 HCs and 17 patients with scrub typhus. Symbols represent individual subjects and horizontal lines are median values. *p

    Article Snippet: Isolation of peripheral blood mononuclear cells (PBMCs) and identification of MAIT cells Peripheral venous blood samples were collected in heparin-containing tubes, and PBMCs were isolated by density-gradient centrifugation using Lymphoprep (Axis-Shield PoC AS, Oslo, Norway).

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

    Decreased circulating MAIT cell numbers in the peripheral blood of scrub typhus patients. Freshly isolated PBMCs from 53 HCs and 38 patients with scrub typhus were stained with APC-Alexa Fluor 750-conjugated anti-CD3, FITC-conjugated anti-TCR γδ, APC-conjugated anti-TCR Vα7.2 and PE-Cy5-conjugated anti-CD161 mAbs, and then analyzed by flow cytometry. Percentages of MAIT cells were calculated using a αβ T cell gate. Panel A : Representative MAIT cell percentages as determined by flow cytometry. Panel B : MAIT cell percentages among peripheral blood αβ T cells. Panel C : Absolute MAIT cell numbers (per microliter of blood). Symbols represent individual subjects and horizontal lines are median values. *p

    Journal: PLoS Neglected Tropical Diseases

    Article Title: Activation, Impaired Tumor Necrosis Factor-α Production, and Deficiency of Circulating Mucosal-Associated Invariant T Cells in Patients with Scrub Typhus

    doi: 10.1371/journal.pntd.0004832

    Figure Lengend Snippet: Decreased circulating MAIT cell numbers in the peripheral blood of scrub typhus patients. Freshly isolated PBMCs from 53 HCs and 38 patients with scrub typhus were stained with APC-Alexa Fluor 750-conjugated anti-CD3, FITC-conjugated anti-TCR γδ, APC-conjugated anti-TCR Vα7.2 and PE-Cy5-conjugated anti-CD161 mAbs, and then analyzed by flow cytometry. Percentages of MAIT cells were calculated using a αβ T cell gate. Panel A : Representative MAIT cell percentages as determined by flow cytometry. Panel B : MAIT cell percentages among peripheral blood αβ T cells. Panel C : Absolute MAIT cell numbers (per microliter of blood). Symbols represent individual subjects and horizontal lines are median values. *p

    Article Snippet: Isolation of peripheral blood mononuclear cells (PBMCs) and identification of MAIT cells Peripheral venous blood samples were collected in heparin-containing tubes, and PBMCs were isolated by density-gradient centrifugation using Lymphoprep (Axis-Shield PoC AS, Oslo, Norway).

    Techniques: Isolation, Staining, Flow Cytometry, Cytometry

    Infection with Mtb HN878 triggers increased IL-22 production through a TLR2-dependent pathway Lung cells isolated from B6, Il1r-/- and Tlr2-/- mice were infected with either Mtb H37Rv (Rv) or Mtb HN878 (HN) at MOI of 0.1 for 7 days, and the protein levels of ( a ) IL-1β and ( b ) IL-22 were assessed in culture supernatants by ELISA and luminex assay, respectively. Lung cells isolated from B6 mice were treated with cell wall extracted from Mtb H37Rv (Rv) or Mtb HN878 (HN), (10μg/ml each). Protein levels of ( c ) IL-1β and ( d ) IL-22 were assessed by ELISA and luminex assay respectively. PBMCs were isolated from ATB patients and were stimulated with Mtb H37Rv (Rv) or Mtb HN878 (HN) cell wall extracts (20 μg/ml each) and the protein levels of ( e ) IL-22 were measured by luminex assays. Error bars represent means ± s.e.m. n = 3-5 for a-d. n = 17 for human ATB patients for e. ND, not detected; UI, uninfected; UN, untreated. * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001 by ( a,b ) one-way ANOVA, ( c,d ) Student's t test ( e ) two-way ANOVA

    Journal: Mucosal immunology

    Article Title: Novel role for IL-22 in protection during chronic Mycobacterium tuberculosis HN878 infection

    doi: 10.1038/mi.2017.15

    Figure Lengend Snippet: Infection with Mtb HN878 triggers increased IL-22 production through a TLR2-dependent pathway Lung cells isolated from B6, Il1r-/- and Tlr2-/- mice were infected with either Mtb H37Rv (Rv) or Mtb HN878 (HN) at MOI of 0.1 for 7 days, and the protein levels of ( a ) IL-1β and ( b ) IL-22 were assessed in culture supernatants by ELISA and luminex assay, respectively. Lung cells isolated from B6 mice were treated with cell wall extracted from Mtb H37Rv (Rv) or Mtb HN878 (HN), (10μg/ml each). Protein levels of ( c ) IL-1β and ( d ) IL-22 were assessed by ELISA and luminex assay respectively. PBMCs were isolated from ATB patients and were stimulated with Mtb H37Rv (Rv) or Mtb HN878 (HN) cell wall extracts (20 μg/ml each) and the protein levels of ( e ) IL-22 were measured by luminex assays. Error bars represent means ± s.e.m. n = 3-5 for a-d. n = 17 for human ATB patients for e. ND, not detected; UI, uninfected; UN, untreated. * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001 by ( a,b ) one-way ANOVA, ( c,d ) Student's t test ( e ) two-way ANOVA

    Article Snippet: Human peripheral blood mononuclear cells (PBMCs) from ATB patients were isolated by Ficoll Hypaque gradient (Lymphoprep; Axis-Shield POC AS, Oslo, Norway ).

    Techniques: Infection, Isolation, Mouse Assay, Enzyme-linked Immunosorbent Assay, Luminex

    Mean (±95% CI ; ±min./max.) peripheral blood mononuclear cell ( PBMC ) nuclear concentration of DNA methyltransferase 3B ( DNMT 3B) following stimulation with pre‐ and post‐exercise plasma. A paired sample t ‐test was used to test significance (* P

    Journal: Physiological Reports

    Article Title: Exercise‐conditioned plasma attenuates nuclear concentrations of DNA methyltransferase 3B in human peripheral blood mononuclear cells. Exercise‐conditioned plasma attenuates nuclear concentrations of DNA methyltransferase 3B in human peripheral blood mononuclear cells

    doi: 10.14814/phy2.12621

    Figure Lengend Snippet: Mean (±95% CI ; ±min./max.) peripheral blood mononuclear cell ( PBMC ) nuclear concentration of DNA methyltransferase 3B ( DNMT 3B) following stimulation with pre‐ and post‐exercise plasma. A paired sample t ‐test was used to test significance (* P

    Article Snippet: The researchers investigated the effect of a 120‐min treadmill run at 60% of v V ˙ O 2 max interspersed with sprints at 90% of v V ˙ O 2 max for the last 30‐sec of every 10‐min (previously shown to induce elevations in plasma IL‐6; Walshe et al. ), followed by a 5‐km time trial, on subsequent changes in DNA methylation of peripheral blood mononuclear cells (PBMCs) as measured by the Infinium Human Methylation 27k beadchip (Illumina, San Diego, CA, USA).

    Techniques: Concentration Assay

    Mean (±95% CI ; ±min./max.) peripheral blood mononuclear cell ( PBMC ) nuclear concentration of DNA methyltransferase 3A ( DNMT 3A) following stimulation with pre‐ and post‐exercise plasma. A paired sample t ‐test was used to test significance.

    Journal: Physiological Reports

    Article Title: Exercise‐conditioned plasma attenuates nuclear concentrations of DNA methyltransferase 3B in human peripheral blood mononuclear cells. Exercise‐conditioned plasma attenuates nuclear concentrations of DNA methyltransferase 3B in human peripheral blood mononuclear cells

    doi: 10.14814/phy2.12621

    Figure Lengend Snippet: Mean (±95% CI ; ±min./max.) peripheral blood mononuclear cell ( PBMC ) nuclear concentration of DNA methyltransferase 3A ( DNMT 3A) following stimulation with pre‐ and post‐exercise plasma. A paired sample t ‐test was used to test significance.

    Article Snippet: The researchers investigated the effect of a 120‐min treadmill run at 60% of v V ˙ O 2 max interspersed with sprints at 90% of v V ˙ O 2 max for the last 30‐sec of every 10‐min (previously shown to induce elevations in plasma IL‐6; Walshe et al. ), followed by a 5‐km time trial, on subsequent changes in DNA methylation of peripheral blood mononuclear cells (PBMCs) as measured by the Infinium Human Methylation 27k beadchip (Illumina, San Diego, CA, USA).

    Techniques: Concentration Assay