human peripheral blood mononuclear cells pbmcs Search Results


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  • 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
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    99
    ATCC primary peripheral blood mononuclear cells pbmc normal human
    S. sonnei induces the secretion of IL-1β, IL-18, NLRP3, ASC, and active caspase-1 in macrophages. (A) J774A.1 macrophages, <t>THP-1</t> macrophages, <t>PBMCs</t> or BMDM were primed with 1 μg/ml LPS for 4 h and then infected with S. sonnei for an additional 20 h. The levels of IL-1β in the supernatants were measured by ELISA. (B–E) J774A.1 macrophages or THP-1 macrophages were primed with 1 μg/ml LPS for 4 h followed and then infected with S. sonnei for an additional 20 h or stimulated with 5 mM ATP for an additional 0.5 h. The levels of IL-1β (B) , IL-18 (C) , caspase-1 (D) , NLRP3, and ASC (E) in the supernatants were measured by Western blotting. The ELISA data are expressed as the mean ± SD of four separate experiments. The Western blotting results are representative of three different experiments. * and *** indicate significant differences at the levels of p
    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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    99
    Millipore histopaque 1077
    γH2AX foci intensity in lymphocytes isolated before (Pre-PET + 0 Gy, Control) and after (Post-PET + 0 Gy, Exposed) a PET scan procedure, as measured by the H2AX Phosphorylation Assay Kit (Flow Cytometry). Mean channel values for quadruplicates were averaged and normalized to an antibody control tube specific to each sample and donor. Error bars represent the standard error of the mean. Lymphocytes were processed for measurement directly following isolation from whole blood using <t>Histopaque-1077</t> and resuspended at 5 × 10 5 cells/mL in complete Rosewell Park Memorial Institute medium. PET indicates positron emission tomography.
    Histopaque 1077, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 13907 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore human peripheral blood mononuclear cells pbmc
    Fusion efficiency enhancement . The fusions were performed according to a standard protocol, where the culture media and PEG were reconstituted from powder forms with either NPD or DI water. For each fusion, <t>PBMC</t> from a single batch were divided into two equal fractures and used to prepare two parallel experiments, in NPD or DI based reagents. The figure presents percent of <t>hybridoma-positive</t> wells in each fusion experiment. The percent was calculated as the number of hybridoma-positive wells from 96-well plates where the cells were seeded and grown after the fusion process. The difference between the NPD- and DI-fusion results was found to be statistically significant by Chi-square analysis (p
    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
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    96
    Millipore ceramide
    Exposure to C1P or <t>ceramide</t> induces P-glycoprotein transport activity at the blood-brain barrier. (A) Dose response of 20 minutes ceramide treatment, showing that ceramide increases specific P-glycoprotein activity in a concentration-dependent manner.
    Ceramide, supplied by Millipore, used in various techniques. Bioz Stars score: 96/100, based on 483 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    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
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    97
    Millipore phytohemagglutinin l
    Effects of VAEI-treated DC on T cells. Purified human CD4 + T lymphocytes were cultured in the presence of medium (NC) or stimulated with <t>phytohemagglutinin-L</t> (PC; 10 μg/ml). Unstimulated T cells were further co-cultivated with immature DC (DC, CD4 + ), DC treated with a maturation cocktail (DC Stim, CD4 + ; 500 ng/mL LPS; 50 ng/mL TNF-alpha and 10 ng/mL IL-1beta), VAEI-treated DC (DC + VAEI, CD4 + ; Iscador® Qu Spez; 0.5 μg/ml), DC incubated with ML-depleted VAEI (DC + VAEI-ML, CD4 + ; Iscador®; 0.66 μg/ml) or VAEI and anti-ML antibody (DC + VAEI + aML-Ab, CD4 + ; 2.5 μg/ml). (A) Cell proliferation analysis was done using CFSE staining and flow cytometry. (B) CD25 surface marker expression was analyzed as a second indicator for T cell activation. Data of 6 individual experiments are presented as mean ± SD in relation to untreated T cells (NC) or untreated T cells co-cultured with immature DC (DC, CD4 + ). (C) IFN-γ release by T cells was analyzed in the supernatants of the co-cultures of 6 independent experiments using a cytokine bead array assay. Asterisks indicate significant differences between the groups (*P
    Phytohemagglutinin L, supplied by Millipore, used in various techniques. Bioz Stars score: 97/100, based on 184 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Axis-Shield Diagnostics human pbmcs
    Human PBMC subsets express receptors for p17. <t>PBMCs</t> were isolated from leukopaks from healthy donors (New York Blood Center) with <t>Lymphoprep.</t> After RBC lysis, cells were stained with p17-Alexa Fluor 430 and surface markers. Live cells were gated as follows:
    Human Pbmcs, supplied by Axis-Shield Diagnostics, used in various techniques. Bioz Stars score: 92/100, based on 140 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    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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    AllCells LLC human pbmcs
    Detection of SMN Protein in SMNA Type 1 <t>PBMCs.</t> SMA Type I patient samples (N = 4) were tested in the SMN ELISA at dilutions of 1∶4 with 1.25×10 6 and 2.5×10 6 PBMCs. SMN protein signal was detected in all samples, with an average of 8.32 pg SMN protein per 10 6 cells. Based on the average of 70.2 pg SMN protein per 10 6 cells calculated for adult normal donor PBMCs, the amount of SMN protein in PBMCs of Type I SMA patients in this patient cohort is 88% less than normal. Error bars represent standard deviations.
    Human Pbmcs, supplied by AllCells LLC, used in various techniques. Bioz Stars score: 92/100, based on 50 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    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
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    99
    Millipore red blood cell lysis buffer
    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
    Red Blood Cell Lysis Buffer, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 3539 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    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
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    93
    Biochrom human 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.
    Human Peripheral Blood Mononuclear Cells Pbmcs, supplied by Biochrom, used in various techniques. Bioz Stars score: 93/100, based on 109 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    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

    S. sonnei induces the secretion of IL-1β, IL-18, NLRP3, ASC, and active caspase-1 in macrophages. (A) J774A.1 macrophages, THP-1 macrophages, PBMCs or BMDM were primed with 1 μg/ml LPS for 4 h and then infected with S. sonnei for an additional 20 h. The levels of IL-1β in the supernatants were measured by ELISA. (B–E) J774A.1 macrophages or THP-1 macrophages were primed with 1 μg/ml LPS for 4 h followed and then infected with S. sonnei for an additional 20 h or stimulated with 5 mM ATP for an additional 0.5 h. The levels of IL-1β (B) , IL-18 (C) , caspase-1 (D) , NLRP3, and ASC (E) in the supernatants were measured by Western blotting. The ELISA data are expressed as the mean ± SD of four separate experiments. The Western blotting results are representative of three different experiments. * and *** indicate significant differences at the levels of p

    Journal: Frontiers in Immunology

    Article Title: Critical Role for the NLRP3 Inflammasome in Mediating IL-1β Production in Shigella sonnei-Infected Macrophages

    doi: 10.3389/fimmu.2020.01115

    Figure Lengend Snippet: S. sonnei induces the secretion of IL-1β, IL-18, NLRP3, ASC, and active caspase-1 in macrophages. (A) J774A.1 macrophages, THP-1 macrophages, PBMCs or BMDM were primed with 1 μg/ml LPS for 4 h and then infected with S. sonnei for an additional 20 h. The levels of IL-1β in the supernatants were measured by ELISA. (B–E) J774A.1 macrophages or THP-1 macrophages were primed with 1 μg/ml LPS for 4 h followed and then infected with S. sonnei for an additional 20 h or stimulated with 5 mM ATP for an additional 0.5 h. The levels of IL-1β (B) , IL-18 (C) , caspase-1 (D) , NLRP3, and ASC (E) in the supernatants were measured by Western blotting. The ELISA data are expressed as the mean ± SD of four separate experiments. The Western blotting results are representative of three different experiments. * and *** indicate significant differences at the levels of p

    Article Snippet: THP-1 macrophages were differentiated from THP-1 monocytes by treatment with 50 nM PMA for 48 h. Human peripheral blood mononuclear cells (PBMCs) were separated from whole blood from healthy volunteers by density gradient centrifugation using Histopaque-1077 , and all experimental protocols were performed in accordance with the guidelines and regulations provided and accepted by the Institutional Review Board of the Tri-Service General Hospital, National Defense Medical Center and the volunteers' informed consent (TSGH-IRB-2-106-05-190 and TSGH-IRB-2-106-05-009).

    Techniques: Infection, Enzyme-linked Immunosorbent Assay, Western Blot

    γH2AX foci intensity in lymphocytes isolated before (Pre-PET + 0 Gy, Control) and after (Post-PET + 0 Gy, Exposed) a PET scan procedure, as measured by the H2AX Phosphorylation Assay Kit (Flow Cytometry). Mean channel values for quadruplicates were averaged and normalized to an antibody control tube specific to each sample and donor. Error bars represent the standard error of the mean. Lymphocytes were processed for measurement directly following isolation from whole blood using Histopaque-1077 and resuspended at 5 × 10 5 cells/mL in complete Rosewell Park Memorial Institute medium. PET indicates positron emission tomography.

    Journal: Dose-Response

    Article Title: Biological Response of Positron Emission Tomography Scan Exposure and Adaptive Response in Humans

    doi: 10.1177/1559325815611904

    Figure Lengend Snippet: γH2AX foci intensity in lymphocytes isolated before (Pre-PET + 0 Gy, Control) and after (Post-PET + 0 Gy, Exposed) a PET scan procedure, as measured by the H2AX Phosphorylation Assay Kit (Flow Cytometry). Mean channel values for quadruplicates were averaged and normalized to an antibody control tube specific to each sample and donor. Error bars represent the standard error of the mean. Lymphocytes were processed for measurement directly following isolation from whole blood using Histopaque-1077 and resuspended at 5 × 10 5 cells/mL in complete Rosewell Park Memorial Institute medium. PET indicates positron emission tomography.

    Article Snippet: Lymphocytes were isolated from whole blood using Histopaque 1077 by centrifugation at room temperature for 30 minutes at 300g , according to the manufacturer’s instructions (Sigma-Aldrich, Oakville, Ontario).

    Techniques: Isolation, Positron Emission Tomography, Phosphorylation Assay, Flow Cytometry, Cytometry

    DGC induces TXNIP down-regulation in T cells. ( A ) Representative Western blot (lower panel) and quantification (upper panel) of TXNIP with CD3ζ as loading control from T cells isolated from blood samples incubated for 4 hours (4 h blood, Procedure II, Fig. 1B ) after no treatment (−) or centrifugation (+). ( B ) Representative Western blot (lower panel) and quantification (upper panel) of TXNIP with CD3ζ as loading control from T cells immediately isolated and lysed (0 h, Procedure I, Fig. 1B ), isolated and incubated for 4 hours before lysis (4 h T cells, Procedure I, Fig. 1B ), and isolated and lysed after incubation of PBMC for 4 hours (4 h PBMC, Procedure III, Fig. 1B ) after DGC on either Lymphoprep, Histopaque or Ficoll-Paque as indicated. ( C ) Representative Western blot (lower panel) and quantification (upper panel) of TXNIP with CD3ζ as loading control from T cells isolated from blood samples incubated for 4 hours (4 h blood, Procedure II, Fig. 1B ) with no addition (Control) or with Lymphoprep, Histopaque or Ficoll-Paque as indicated. ( D ) Overview of different T cell isolation and incubation procedures used. The text in the red frames gives the procedure used and the nomenclature for the T cell lysates analyzed and shown in ( E) lanes 4–9. ( E ) Representative Western blot (lower panel) and quantification (upper panel) of TXNIP with CD3ζ as loading control from T cells immediately isolated and lysed (lane 1: 0 h, Procedure I, Fig. 1B ; lane 4: monocyte depletion, 0 h, Procedure IV, ( D) ; lane 7: T cell enrichment, 0 h, Procedure VI, D ), isolated and incubated for 4 hours before lysis (lane 2: 4 h T cells, Procedure I, Fig. 1B ; lane 5: monocyte depletion, 4 h T cells, Procedure IV, ( D) ; lane 8: T cell enrichment, 4 h T cells, Procedure VI, D ), and isolated and lysed after incubation of PBMC for 4 hours (lane 3: 4 h PBMC, Procedure III, Fig. 1B ; lane 6: monocyte depletion, 4 h PBMC, Procedure V, D ) or T cells for 5 h (lane 9: T cell enrichment, 5 h T cells, Procedure VI, D ). ( A–C , E ) Each Western blot is representative for Western blots obtained from at least 3 different biological experiments and the quantification shows the mean + SEM of the band densities of TXNIP from Western blots obtained from at least 3 different biological experiments. The positions of the relevant molecular weight markers and their molecular weight in kDa are given to the right of each Western blot.

    Journal: Scientific Reports

    Article Title: Tumor necrosis factor induces rapid down-regulation of TXNIP in human T cells

    doi: 10.1038/s41598-019-53234-x

    Figure Lengend Snippet: DGC induces TXNIP down-regulation in T cells. ( A ) Representative Western blot (lower panel) and quantification (upper panel) of TXNIP with CD3ζ as loading control from T cells isolated from blood samples incubated for 4 hours (4 h blood, Procedure II, Fig. 1B ) after no treatment (−) or centrifugation (+). ( B ) Representative Western blot (lower panel) and quantification (upper panel) of TXNIP with CD3ζ as loading control from T cells immediately isolated and lysed (0 h, Procedure I, Fig. 1B ), isolated and incubated for 4 hours before lysis (4 h T cells, Procedure I, Fig. 1B ), and isolated and lysed after incubation of PBMC for 4 hours (4 h PBMC, Procedure III, Fig. 1B ) after DGC on either Lymphoprep, Histopaque or Ficoll-Paque as indicated. ( C ) Representative Western blot (lower panel) and quantification (upper panel) of TXNIP with CD3ζ as loading control from T cells isolated from blood samples incubated for 4 hours (4 h blood, Procedure II, Fig. 1B ) with no addition (Control) or with Lymphoprep, Histopaque or Ficoll-Paque as indicated. ( D ) Overview of different T cell isolation and incubation procedures used. The text in the red frames gives the procedure used and the nomenclature for the T cell lysates analyzed and shown in ( E) lanes 4–9. ( E ) Representative Western blot (lower panel) and quantification (upper panel) of TXNIP with CD3ζ as loading control from T cells immediately isolated and lysed (lane 1: 0 h, Procedure I, Fig. 1B ; lane 4: monocyte depletion, 0 h, Procedure IV, ( D) ; lane 7: T cell enrichment, 0 h, Procedure VI, D ), isolated and incubated for 4 hours before lysis (lane 2: 4 h T cells, Procedure I, Fig. 1B ; lane 5: monocyte depletion, 4 h T cells, Procedure IV, ( D) ; lane 8: T cell enrichment, 4 h T cells, Procedure VI, D ), and isolated and lysed after incubation of PBMC for 4 hours (lane 3: 4 h PBMC, Procedure III, Fig. 1B ; lane 6: monocyte depletion, 4 h PBMC, Procedure V, D ) or T cells for 5 h (lane 9: T cell enrichment, 5 h T cells, Procedure VI, D ). ( A–C , E ) Each Western blot is representative for Western blots obtained from at least 3 different biological experiments and the quantification shows the mean + SEM of the band densities of TXNIP from Western blots obtained from at least 3 different biological experiments. The positions of the relevant molecular weight markers and their molecular weight in kDa are given to the right of each Western blot.

    Article Snippet: Where indicated, Histopaque® –1077 (10771, Sigma-Aldrich) or Ficoll-PaqueTM (GE17-5442-02, Sigma-Aldrich) was used and centrifuged for 30 min at 400 g, 24 °C with the brake off as specified by the manufacturers’ instructions.

    Techniques: Western Blot, Isolation, Incubation, Centrifugation, Lysis, Cell Isolation, Molecular Weight

    Fusion efficiency enhancement . The fusions were performed according to a standard protocol, where the culture media and PEG were reconstituted from powder forms with either NPD or DI water. For each fusion, PBMC from a single batch were divided into two equal fractures and used to prepare two parallel experiments, in NPD or DI based reagents. The figure presents percent of hybridoma-positive wells in each fusion experiment. The percent was calculated as the number of hybridoma-positive wells from 96-well plates where the cells were seeded and grown after the fusion process. The difference between the NPD- and DI-fusion results was found to be statistically significant by Chi-square analysis (p

    Journal: BMC Biotechnology

    Article Title: Enhancement of hybridoma formation, clonability and cell proliferation in a nanoparticle-doped aqueous environment

    doi: 10.1186/1472-6750-8-3

    Figure Lengend Snippet: Fusion efficiency enhancement . The fusions were performed according to a standard protocol, where the culture media and PEG were reconstituted from powder forms with either NPD or DI water. For each fusion, PBMC from a single batch were divided into two equal fractures and used to prepare two parallel experiments, in NPD or DI based reagents. The figure presents percent of hybridoma-positive wells in each fusion experiment. The percent was calculated as the number of hybridoma-positive wells from 96-well plates where the cells were seeded and grown after the fusion process. The difference between the NPD- and DI-fusion results was found to be statistically significant by Chi-square analysis (p

    Article Snippet: For production of hybridoma cells, human peripheral blood mononuclear cells (PBMC) were isolated from 40 mL of freshly drawn whole blood, purified with Histopaque 1077 (Sigma) as previously described, and washed 4 times in DI based culture medium without serum.

    Techniques:

    Exposure to C1P or ceramide induces P-glycoprotein transport activity at the blood-brain barrier. (A) Dose response of 20 minutes ceramide treatment, showing that ceramide increases specific P-glycoprotein activity in a concentration-dependent manner.

    Journal: Molecular Pharmacology

    Article Title:

    doi: 10.1124/mol.116.107169

    Figure Lengend Snippet: Exposure to C1P or ceramide induces P-glycoprotein transport activity at the blood-brain barrier. (A) Dose response of 20 minutes ceramide treatment, showing that ceramide increases specific P-glycoprotein activity in a concentration-dependent manner.

    Article Snippet: Ceramide from bovine spinal cord, mouse monoclonal β -actin antibody, chlorpromazine hydrochloride, Ficoll PM 400, cycloheximide, SC-51089 hydrate [3-chloro- N ʹ-(3-pyridin-4-ylpropanoyl)-6 H -benzo[ b ][1,4]benzoxazepine-5-carbohydrazide;hydrate;hydrochloride], PF-04418948 [1-(4-fluorobenzoyl)-3-[(6-methoxynaphthalen-2-yl)oxymethyl]azetidine-3-carboxylic acid], celecoxib, NVP-231 [ N -(2-benzamido-1,3-benzothiazol-6-yl)adamantane-1-carboxamide], and all other chemicals were obtained from Sigma-Aldrich (St. Louis, MO).

    Techniques: Activity Assay, Concentration Assay

    Proposed signaling cascade for the induction of P-glycoprotein activity by ceramide and C1P.

    Journal: Molecular Pharmacology

    Article Title:

    doi: 10.1124/mol.116.107169

    Figure Lengend Snippet: Proposed signaling cascade for the induction of P-glycoprotein activity by ceramide and C1P.

    Article Snippet: Ceramide from bovine spinal cord, mouse monoclonal β -actin antibody, chlorpromazine hydrochloride, Ficoll PM 400, cycloheximide, SC-51089 hydrate [3-chloro- N ʹ-(3-pyridin-4-ylpropanoyl)-6 H -benzo[ b ][1,4]benzoxazepine-5-carbohydrazide;hydrate;hydrochloride], PF-04418948 [1-(4-fluorobenzoyl)-3-[(6-methoxynaphthalen-2-yl)oxymethyl]azetidine-3-carboxylic acid], celecoxib, NVP-231 [ N -(2-benzamido-1,3-benzothiazol-6-yl)adamantane-1-carboxamide], and all other chemicals were obtained from Sigma-Aldrich (St. Louis, MO).

    Techniques: Activity Assay

    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

    Effects of VAEI-treated DC on T cells. Purified human CD4 + T lymphocytes were cultured in the presence of medium (NC) or stimulated with phytohemagglutinin-L (PC; 10 μg/ml). Unstimulated T cells were further co-cultivated with immature DC (DC, CD4 + ), DC treated with a maturation cocktail (DC Stim, CD4 + ; 500 ng/mL LPS; 50 ng/mL TNF-alpha and 10 ng/mL IL-1beta), VAEI-treated DC (DC + VAEI, CD4 + ; Iscador® Qu Spez; 0.5 μg/ml), DC incubated with ML-depleted VAEI (DC + VAEI-ML, CD4 + ; Iscador®; 0.66 μg/ml) or VAEI and anti-ML antibody (DC + VAEI + aML-Ab, CD4 + ; 2.5 μg/ml). (A) Cell proliferation analysis was done using CFSE staining and flow cytometry. (B) CD25 surface marker expression was analyzed as a second indicator for T cell activation. Data of 6 individual experiments are presented as mean ± SD in relation to untreated T cells (NC) or untreated T cells co-cultured with immature DC (DC, CD4 + ). (C) IFN-γ release by T cells was analyzed in the supernatants of the co-cultures of 6 independent experiments using a cytokine bead array assay. Asterisks indicate significant differences between the groups (*P

    Journal: PLoS ONE

    Article Title: Viscum album neutralizes tumor-induced immunosuppression in a human in vitro cell model

    doi: 10.1371/journal.pone.0181553

    Figure Lengend Snippet: Effects of VAEI-treated DC on T cells. Purified human CD4 + T lymphocytes were cultured in the presence of medium (NC) or stimulated with phytohemagglutinin-L (PC; 10 μg/ml). Unstimulated T cells were further co-cultivated with immature DC (DC, CD4 + ), DC treated with a maturation cocktail (DC Stim, CD4 + ; 500 ng/mL LPS; 50 ng/mL TNF-alpha and 10 ng/mL IL-1beta), VAEI-treated DC (DC + VAEI, CD4 + ; Iscador® Qu Spez; 0.5 μg/ml), DC incubated with ML-depleted VAEI (DC + VAEI-ML, CD4 + ; Iscador®; 0.66 μg/ml) or VAEI and anti-ML antibody (DC + VAEI + aML-Ab, CD4 + ; 2.5 μg/ml). (A) Cell proliferation analysis was done using CFSE staining and flow cytometry. (B) CD25 surface marker expression was analyzed as a second indicator for T cell activation. Data of 6 individual experiments are presented as mean ± SD in relation to untreated T cells (NC) or untreated T cells co-cultured with immature DC (DC, CD4 + ). (C) IFN-γ release by T cells was analyzed in the supernatants of the co-cultures of 6 independent experiments using a cytokine bead array assay. Asterisks indicate significant differences between the groups (*P

    Article Snippet: As controls, CFSE+ CD4+ T cells were cultured with medium alone (NC) or in the presence of phytohemagglutinin-L (PC; PHA-L; 10 μg/mL; Sigma-Aldrich, Taufkirchen, Germany).

    Techniques: Purification, Cell Culture, Incubation, Staining, Flow Cytometry, Cytometry, Marker, Expressing, Activation Assay

    Effects of VAEI-treated DC on T cells after tumor-induced immunosuppression. Purified human CD4 + T lymphocytes were cultured in the presence of medium (NC) or stimulated with phytohemagglutinin-L (PC; 10 μg/ml). Co-cultivation of T cells was carried out with unstimulated DC treated with 10% RPMI 1640 medium (CD4 + + DC + Medium), 10% tumor supernatant of an RCC line (CD4 + + DC + TS + ), TS and VAEI (CD4 + + DC + TS + VAEI; Iscador® Qu Spez; 0.5 μg/ml), TS and ML-depleted VAEI (CD4 + + DC + TS + VAEI-ML; Iscador®; 0.66 μg/ml) or TS, VAEI and anti-ML antibody (CD4 + + DC + TS + VAEI + aML-Ab, CD4 + ; 2.5 μg/ml). (A) Cell division analysis was done using CFSE staining and flow cytometry. (B) CD25 surface marker expression was analyzed as a second indicator for T cell activation. Data from 6 individual experiments are presented as mean ± SD in relation to untreated T cells (NC) or T cells co-cultured with TS-treated DC (DC + TS). (C) IFN-γ release by T cells was analyzed in the supernatants of 6 independent co-culture experiments using cytokine bead array assay. Asterisks indicate significant differences between the groups (*P

    Journal: PLoS ONE

    Article Title: Viscum album neutralizes tumor-induced immunosuppression in a human in vitro cell model

    doi: 10.1371/journal.pone.0181553

    Figure Lengend Snippet: Effects of VAEI-treated DC on T cells after tumor-induced immunosuppression. Purified human CD4 + T lymphocytes were cultured in the presence of medium (NC) or stimulated with phytohemagglutinin-L (PC; 10 μg/ml). Co-cultivation of T cells was carried out with unstimulated DC treated with 10% RPMI 1640 medium (CD4 + + DC + Medium), 10% tumor supernatant of an RCC line (CD4 + + DC + TS + ), TS and VAEI (CD4 + + DC + TS + VAEI; Iscador® Qu Spez; 0.5 μg/ml), TS and ML-depleted VAEI (CD4 + + DC + TS + VAEI-ML; Iscador®; 0.66 μg/ml) or TS, VAEI and anti-ML antibody (CD4 + + DC + TS + VAEI + aML-Ab, CD4 + ; 2.5 μg/ml). (A) Cell division analysis was done using CFSE staining and flow cytometry. (B) CD25 surface marker expression was analyzed as a second indicator for T cell activation. Data from 6 individual experiments are presented as mean ± SD in relation to untreated T cells (NC) or T cells co-cultured with TS-treated DC (DC + TS). (C) IFN-γ release by T cells was analyzed in the supernatants of 6 independent co-culture experiments using cytokine bead array assay. Asterisks indicate significant differences between the groups (*P

    Article Snippet: As controls, CFSE+ CD4+ T cells were cultured with medium alone (NC) or in the presence of phytohemagglutinin-L (PC; PHA-L; 10 μg/mL; Sigma-Aldrich, Taufkirchen, Germany).

    Techniques: Purification, Cell Culture, Staining, Flow Cytometry, Cytometry, Marker, Expressing, Activation Assay, Co-Culture Assay

    Human PBMC subsets express receptors for p17. PBMCs were isolated from leukopaks from healthy donors (New York Blood Center) with Lymphoprep. After RBC lysis, cells were stained with p17-Alexa Fluor 430 and surface markers. Live cells were gated as follows:

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

    Article Title: Expression of HIV-1 matrix protein p17 and association with B-cell lymphoma in HIV-1 transgenic mice

    doi: 10.1073/pnas.1615258113

    Figure Lengend Snippet: Human PBMC subsets express receptors for p17. PBMCs were isolated from leukopaks from healthy donors (New York Blood Center) with Lymphoprep. After RBC lysis, cells were stained with p17-Alexa Fluor 430 and surface markers. Live cells were gated as follows:

    Article Snippet: Human PBMCs were isolated using Lymphoprep (Axis-Shield) from healthy blood donors' leukopaks obtained from the New York Blood Center, Long Island, NY, in accordance with their guidelines and those of the Institutional Review Board of the University of Maryland School of Medicine.

    Techniques: Isolation, Lysis, Staining

    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

    Detection of SMN Protein in SMNA Type 1 PBMCs. SMA Type I patient samples (N = 4) were tested in the SMN ELISA at dilutions of 1∶4 with 1.25×10 6 and 2.5×10 6 PBMCs. SMN protein signal was detected in all samples, with an average of 8.32 pg SMN protein per 10 6 cells. Based on the average of 70.2 pg SMN protein per 10 6 cells calculated for adult normal donor PBMCs, the amount of SMN protein in PBMCs of Type I SMA patients in this patient cohort is 88% less than normal. Error bars represent standard deviations.

    Journal: PLoS ONE

    Article Title: Utility of Survival Motor Neuron ELISA for Spinal Muscular Atrophy Clinical and Preclinical Analyses

    doi: 10.1371/journal.pone.0024269

    Figure Lengend Snippet: Detection of SMN Protein in SMNA Type 1 PBMCs. SMA Type I patient samples (N = 4) were tested in the SMN ELISA at dilutions of 1∶4 with 1.25×10 6 and 2.5×10 6 PBMCs. SMN protein signal was detected in all samples, with an average of 8.32 pg SMN protein per 10 6 cells. Based on the average of 70.2 pg SMN protein per 10 6 cells calculated for adult normal donor PBMCs, the amount of SMN protein in PBMCs of Type I SMA patients in this patient cohort is 88% less than normal. Error bars represent standard deviations.

    Article Snippet: Non-human primate PBMC lysates were run alongside lysates from normal human PBMCs (from AllCells) and mouse PBMCs harvested from 8 week old FVBn animals at PharmOptima.

    Techniques: Enzyme-linked Immunosorbent Assay

    Quantitation of SMN by PBMC subtype. There was no significant difference in SMN protein levels between human lymphocytes and monocytes fractions separated from PBMCs. SMN protein was detectable in the majority of samples for both cell types, ranging from 6.8–25.8 pg/10 6 cells for lymphocytes and 8–17.5 pg/10 6 cells for monocytes. Error bars represent standard deviations.

    Journal: PLoS ONE

    Article Title: Utility of Survival Motor Neuron ELISA for Spinal Muscular Atrophy Clinical and Preclinical Analyses

    doi: 10.1371/journal.pone.0024269

    Figure Lengend Snippet: Quantitation of SMN by PBMC subtype. There was no significant difference in SMN protein levels between human lymphocytes and monocytes fractions separated from PBMCs. SMN protein was detectable in the majority of samples for both cell types, ranging from 6.8–25.8 pg/10 6 cells for lymphocytes and 8–17.5 pg/10 6 cells for monocytes. Error bars represent standard deviations.

    Article Snippet: Non-human primate PBMC lysates were run alongside lysates from normal human PBMCs (from AllCells) and mouse PBMCs harvested from 8 week old FVBn animals at PharmOptima.

    Techniques: Quantitation Assay

    ELISA Performance, parallelism and detection of SMN in human PBMCs. A : The recombinant hSMN standard curve data developed from N = 6 curves were highly reproducible with standard deviations of about ±0.23 OD unit variations. The dotted lines surrounding the dose-response curve represent 2 standard deviations. B : Comparison of SMN signal detection between recombinant human SMN (dilutions from 1∶4 to 1∶256) and SMN extracted from HeLa cell lysates (1∶4–1∶32) revealed a high degree of parallelism between reagents, allowing for accurate evaluation of native SMN protein using the recombinant SMN ELISA standard. C : The SMN ELISA detected protein in adult donor PBMCs; values ranged from 86 to 229 pg SMN per well at a dilution of 1∶4 and an average of 70.2 pg SMN protein per 10 6 cells. 10 7 PBMCs diluted 1∶4 through 1∶32 produced linear SMN protein levels. Even with only N = 6 normal samples, SMN levels in PBMCs varied by nearly 3-fold. Error bars represent standard deviations.

    Journal: PLoS ONE

    Article Title: Utility of Survival Motor Neuron ELISA for Spinal Muscular Atrophy Clinical and Preclinical Analyses

    doi: 10.1371/journal.pone.0024269

    Figure Lengend Snippet: ELISA Performance, parallelism and detection of SMN in human PBMCs. A : The recombinant hSMN standard curve data developed from N = 6 curves were highly reproducible with standard deviations of about ±0.23 OD unit variations. The dotted lines surrounding the dose-response curve represent 2 standard deviations. B : Comparison of SMN signal detection between recombinant human SMN (dilutions from 1∶4 to 1∶256) and SMN extracted from HeLa cell lysates (1∶4–1∶32) revealed a high degree of parallelism between reagents, allowing for accurate evaluation of native SMN protein using the recombinant SMN ELISA standard. C : The SMN ELISA detected protein in adult donor PBMCs; values ranged from 86 to 229 pg SMN per well at a dilution of 1∶4 and an average of 70.2 pg SMN protein per 10 6 cells. 10 7 PBMCs diluted 1∶4 through 1∶32 produced linear SMN protein levels. Even with only N = 6 normal samples, SMN levels in PBMCs varied by nearly 3-fold. Error bars represent standard deviations.

    Article Snippet: Non-human primate PBMC lysates were run alongside lysates from normal human PBMCs (from AllCells) and mouse PBMCs harvested from 8 week old FVBn animals at PharmOptima.

    Techniques: Enzyme-linked Immunosorbent Assay, Recombinant, Produced

    Smn protein levels by tissue in WT mice. Smn protein levels are distinct across the an array of tissues and regions of the brain. A : Smn levels varied by as much as 10-fold across tissue types in adult FVB mice (14 weeks old). SMN levels by tissue were distributed on a basis loosely ordered by having lesser to greater dividing cell populations. Smn levels for tissues are represented as Smn pg/mg total protein. PBMCs are represented as Smn pg/mg total protein in the lysis buffer extract. On a per cell basis the average Smn level was 67.2 pg/10 6 PBMCs. B : Smn protein in the brain, spinal cord, sciatic nerve and quadriceps muscle of wildtype mice shows marked variations, with spinal cord having 6-fold greater levels than muscle and nerve. In the brain, Smn protein signal in the thalamus was nearly 2.5-fold less than hippocampal levels. Error bars represent standard deviations.

    Journal: PLoS ONE

    Article Title: Utility of Survival Motor Neuron ELISA for Spinal Muscular Atrophy Clinical and Preclinical Analyses

    doi: 10.1371/journal.pone.0024269

    Figure Lengend Snippet: Smn protein levels by tissue in WT mice. Smn protein levels are distinct across the an array of tissues and regions of the brain. A : Smn levels varied by as much as 10-fold across tissue types in adult FVB mice (14 weeks old). SMN levels by tissue were distributed on a basis loosely ordered by having lesser to greater dividing cell populations. Smn levels for tissues are represented as Smn pg/mg total protein. PBMCs are represented as Smn pg/mg total protein in the lysis buffer extract. On a per cell basis the average Smn level was 67.2 pg/10 6 PBMCs. B : Smn protein in the brain, spinal cord, sciatic nerve and quadriceps muscle of wildtype mice shows marked variations, with spinal cord having 6-fold greater levels than muscle and nerve. In the brain, Smn protein signal in the thalamus was nearly 2.5-fold less than hippocampal levels. Error bars represent standard deviations.

    Article Snippet: Non-human primate PBMC lysates were run alongside lysates from normal human PBMCs (from AllCells) and mouse PBMCs harvested from 8 week old FVBn animals at PharmOptima.

    Techniques: Mouse Assay, Lysis

    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:

    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