characterization mononuclear cells mncs  (Millipore)


Bioz Verified Symbol Millipore is a verified supplier  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99

    Structured Review

    Millipore characterization mononuclear cells mncs
    Characterization of isolated <t>EPCs.</t> (A) Bone marrow derived <t>MNCs</t> appear a spindle-shape and cluster arrangement after 7-day culture under endothelial-specific conditions; the scale bars represent 200 μm. (B) DiI-acLDL and FITC-UEA-1 uptake assay show that the cells are both DiI-acLDL/FITC-UEA-1 + , which indicates that the isolated MNCs are EPCs, the scale bars represent 50 μm. (C) The cultured MNCs were further characterized by immunofluorescent staining using EPCs specific markers CD133/VEGFR-2 and nuclear maker 4′,6-diamidino-2-phenylindole (DAPI); the scale bars represent 50 μm.
    Characterization Mononuclear Cells Mncs, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 18 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/characterization mononuclear cells mncs/product/Millipore
    Average 99 stars, based on 18 article reviews
    Price from $9.99 to $1999.99
    characterization mononuclear cells mncs - by Bioz Stars, 2020-09
    99/100 stars

    Images

    1) Product Images from "The role of CXCR7 on the adhesion, proliferation and angiogenesis of endothelial progenitor cells"

    Article Title: The role of CXCR7 on the adhesion, proliferation and angiogenesis of endothelial progenitor cells

    Journal: Journal of Cellular and Molecular Medicine

    doi: 10.1111/j.1582-4934.2011.01301.x

    Characterization of isolated EPCs. (A) Bone marrow derived MNCs appear a spindle-shape and cluster arrangement after 7-day culture under endothelial-specific conditions; the scale bars represent 200 μm. (B) DiI-acLDL and FITC-UEA-1 uptake assay show that the cells are both DiI-acLDL/FITC-UEA-1 + , which indicates that the isolated MNCs are EPCs, the scale bars represent 50 μm. (C) The cultured MNCs were further characterized by immunofluorescent staining using EPCs specific markers CD133/VEGFR-2 and nuclear maker 4′,6-diamidino-2-phenylindole (DAPI); the scale bars represent 50 μm.
    Figure Legend Snippet: Characterization of isolated EPCs. (A) Bone marrow derived MNCs appear a spindle-shape and cluster arrangement after 7-day culture under endothelial-specific conditions; the scale bars represent 200 μm. (B) DiI-acLDL and FITC-UEA-1 uptake assay show that the cells are both DiI-acLDL/FITC-UEA-1 + , which indicates that the isolated MNCs are EPCs, the scale bars represent 50 μm. (C) The cultured MNCs were further characterized by immunofluorescent staining using EPCs specific markers CD133/VEGFR-2 and nuclear maker 4′,6-diamidino-2-phenylindole (DAPI); the scale bars represent 50 μm.

    Techniques Used: Isolation, Derivative Assay, Cell Culture, Staining

    Related Articles

    Centrifugation:

    Article Title: Neither Lys- and DAP-type peptidoglycans stimulate mouse or human innate immune cells via Toll-like receptor 2
    Article Snippet: .. Assay of biological activity of PGN in human monocytes Peripheral blood mononuclear cells (PBMC) were prepared from heparinized blood by centrifugation through Histopaque 1077 (Sigma) according to the manufacturers protocol. .. PBMCs were washed, resupended in RPMI supplemented with 1% FBS and 0.2 mg/ml human IgG, and stimulated for 12 hours in the presence of Brefeldin A (3 μg/ml) with 10 μg/ml PGN of the initial SDS extraction, after HF treatment and PGN after final purification.

    Article Title: Cyclosporin A-Sensitive Transcription Factor Egr-3 Regulates Fas Ligand Expression
    Article Snippet: .. To generate human T-cell blasts, human peripheral blood mononuclear cells were isolated by density centrifugation with the use of a lymphocyte separation medium (Biofluids) cultured in complete medium for 2 days with 2 ng of phorbol myristate acetate (PMA) (Sigma) per ml and 1 μg of ionomycin (Sigma) per ml, washed, and cultured for an additional day with 10 U of recombinant human IL-2 (Cetus/Chiron, Emeryville, Calif.) per ml, provided by J. Wunderlich (National Institutes of Health). .. A 1.2-kb human fasL upstream genomic region fragment was isolated with the Promoterfinder PCR-based kit from Promega Corp. (Madison, Wis.) and cloned into the luciferase reporter construct pGL3 (Promega) to create the construct 1.2-FasL-GL3.

    Gradient Centrifugation:

    Article Title: Finasteride Enhances the Generation of Human Myeloid-Derived Suppressor Cells by Up-Regulating the COX2/PGE2 Pathway
    Article Snippet: .. PBMC isolation Human peripheral blood mononuclear cells (PBMCs) were isolated from healthy volunteer donors by density gradient centrifugation through Ficoll-Hypaque (Sigma-Aldrich, St. Louis, MO), following the procedures described by our group previously [ ]. .. Purified PBMCs were analyzed or treated immediately, and cultured at 37°C in the conditioned RPMI 1640 (Gibco) supplemented with 10% fetal bovine serum (Gibco), 2 mM L-glutamine (Gibco), 100 U/ml penicillin (Gibco), and 100 μg/ml streptomycin (Gibco) in humidified 5% CO2 incubators.

    Article Title: Novel oligodeoxynucleotide agonists of TLR9 containing N3-Me-dC or N1-Me-dG modifications
    Article Snippet: .. Human PBMC isolation Peripheral blood mononuclear cells (PBMCs) from freshly drawn healthy volunteer blood (Research Blood Components, Brighton, MA) were isolated by Ficoll density gradient centrifugation (Histopaque-1077, Sigma). .. Human cytokine ELISAs Human PBMCs were plated in 96-well plates at a concentration of 5 × 106 cells/ml.

    Article Title: α-Lactosylceramide Protects Against iNKT-Mediated Murine Airway Hyperreactivity and Liver Injury Through Competitive Inhibition of Cd1d Binding
    Article Snippet: .. In vitro Culture of Human PBMCs and iNKT Cells Human peripheral blood mononuclear cells (PBMCs) from healthy volunteers were collected from whole blood through gradient-centrifugation with Histopaque-1077 (Sigma-Aldrich). .. Human iNKT cells were isolated from PBMCs as described previously (Kim et al., ).

    Article Title: Fucoidan inhibits CCL22 production through NF-κB pathway in M2 macrophages: a potential therapeutic strategy for cancer
    Article Snippet: .. Preparation and isolation of PBMC and cell subtypes Human peripheral blood mononuclear cells (PBMCs) were isolated from leukocyte-enriched buffy coats of healthy donors by Ficoll (Sigma-Aldrich) density gradient centrifugation. ..

    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 *
    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). .. These cells were cultured for 7 days in the presence of M-CSF (100 ng/ml).

    In Vitro:

    Article Title: α-Lactosylceramide Protects Against iNKT-Mediated Murine Airway Hyperreactivity and Liver Injury Through Competitive Inhibition of Cd1d Binding
    Article Snippet: .. In vitro Culture of Human PBMCs and iNKT Cells Human peripheral blood mononuclear cells (PBMCs) from healthy volunteers were collected from whole blood through gradient-centrifugation with Histopaque-1077 (Sigma-Aldrich). .. Human iNKT cells were isolated from PBMCs as described previously (Kim et al., ).

    Isolation:

    Article Title: Finasteride Enhances the Generation of Human Myeloid-Derived Suppressor Cells by Up-Regulating the COX2/PGE2 Pathway
    Article Snippet: .. PBMC isolation Human peripheral blood mononuclear cells (PBMCs) were isolated from healthy volunteer donors by density gradient centrifugation through Ficoll-Hypaque (Sigma-Aldrich, St. Louis, MO), following the procedures described by our group previously [ ]. .. Purified PBMCs were analyzed or treated immediately, and cultured at 37°C in the conditioned RPMI 1640 (Gibco) supplemented with 10% fetal bovine serum (Gibco), 2 mM L-glutamine (Gibco), 100 U/ml penicillin (Gibco), and 100 μg/ml streptomycin (Gibco) in humidified 5% CO2 incubators.

    Article Title: Novel oligodeoxynucleotide agonists of TLR9 containing N3-Me-dC or N1-Me-dG modifications
    Article Snippet: .. Human PBMC isolation Peripheral blood mononuclear cells (PBMCs) from freshly drawn healthy volunteer blood (Research Blood Components, Brighton, MA) were isolated by Ficoll density gradient centrifugation (Histopaque-1077, Sigma). .. Human cytokine ELISAs Human PBMCs were plated in 96-well plates at a concentration of 5 × 106 cells/ml.

    Article Title: Fucoidan inhibits CCL22 production through NF-κB pathway in M2 macrophages: a potential therapeutic strategy for cancer
    Article Snippet: .. Preparation and isolation of PBMC and cell subtypes Human peripheral blood mononuclear cells (PBMCs) were isolated from leukocyte-enriched buffy coats of healthy donors by Ficoll (Sigma-Aldrich) density gradient centrifugation. ..

    Article Title: Cyclosporin A-Sensitive Transcription Factor Egr-3 Regulates Fas Ligand Expression
    Article Snippet: .. To generate human T-cell blasts, human peripheral blood mononuclear cells were isolated by density centrifugation with the use of a lymphocyte separation medium (Biofluids) cultured in complete medium for 2 days with 2 ng of phorbol myristate acetate (PMA) (Sigma) per ml and 1 μg of ionomycin (Sigma) per ml, washed, and cultured for an additional day with 10 U of recombinant human IL-2 (Cetus/Chiron, Emeryville, Calif.) per ml, provided by J. Wunderlich (National Institutes of Health). .. A 1.2-kb human fasL upstream genomic region fragment was isolated with the Promoterfinder PCR-based kit from Promega Corp. (Madison, Wis.) and cloned into the luciferase reporter construct pGL3 (Promega) to create the construct 1.2-FasL-GL3.

    Article Title: Combining flagellin and human β-defensin-3 to combat bacterial infections
    Article Snippet: .. ISOLATION OF PERIPHERAL BLOOD MONONUCLEAR CELLS (PBMCs) FROM HEALTHY DONORS The human peripheral blood mononuclear cells (PBMCs) from healthy donors were isolated on a Ficoll–Isopaque gradient (d 1.077 Sigma-Aldrich, St. Louis, MO, USA). .. The isolated PBMCs were then counted and checked for viability using Trypan blue.

    Cell Culture:

    Article Title: Cyclosporin A-Sensitive Transcription Factor Egr-3 Regulates Fas Ligand Expression
    Article Snippet: .. To generate human T-cell blasts, human peripheral blood mononuclear cells were isolated by density centrifugation with the use of a lymphocyte separation medium (Biofluids) cultured in complete medium for 2 days with 2 ng of phorbol myristate acetate (PMA) (Sigma) per ml and 1 μg of ionomycin (Sigma) per ml, washed, and cultured for an additional day with 10 U of recombinant human IL-2 (Cetus/Chiron, Emeryville, Calif.) per ml, provided by J. Wunderlich (National Institutes of Health). .. A 1.2-kb human fasL upstream genomic region fragment was isolated with the Promoterfinder PCR-based kit from Promega Corp. (Madison, Wis.) and cloned into the luciferase reporter construct pGL3 (Promega) to create the construct 1.2-FasL-GL3.

    Activity Assay:

    Article Title: Neither Lys- and DAP-type peptidoglycans stimulate mouse or human innate immune cells via Toll-like receptor 2
    Article Snippet: .. Assay of biological activity of PGN in human monocytes Peripheral blood mononuclear cells (PBMC) were prepared from heparinized blood by centrifugation through Histopaque 1077 (Sigma) according to the manufacturers protocol. .. PBMCs were washed, resupended in RPMI supplemented with 1% FBS and 0.2 mg/ml human IgG, and stimulated for 12 hours in the presence of Brefeldin A (3 μg/ml) with 10 μg/ml PGN of the initial SDS extraction, after HF treatment and PGN after final purification.

    Recombinant:

    Article Title: Cyclosporin A-Sensitive Transcription Factor Egr-3 Regulates Fas Ligand Expression
    Article Snippet: .. To generate human T-cell blasts, human peripheral blood mononuclear cells were isolated by density centrifugation with the use of a lymphocyte separation medium (Biofluids) cultured in complete medium for 2 days with 2 ng of phorbol myristate acetate (PMA) (Sigma) per ml and 1 μg of ionomycin (Sigma) per ml, washed, and cultured for an additional day with 10 U of recombinant human IL-2 (Cetus/Chiron, Emeryville, Calif.) per ml, provided by J. Wunderlich (National Institutes of Health). .. A 1.2-kb human fasL upstream genomic region fragment was isolated with the Promoterfinder PCR-based kit from Promega Corp. (Madison, Wis.) and cloned into the luciferase reporter construct pGL3 (Promega) to create the construct 1.2-FasL-GL3.

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 89
    Millipore culture pbmcs
    Features of ssRNA40-induced LILRA3 expression. a Kinetics of LILRA3 expression was compared to other ssRNA40 inducible cytokines. <t>PBMCs</t> from two donors were stimulated with ssRNA40 or ssRNA41 and the RNA was harvested at various time points. qPCR for LILRA3 , TNF , IFNb , IFNg , and IL1b expression, normalized to RPLP0 and relatively quantified to calibrator cDNA (pooled cDNA from 10. b Effect of cytokine inhibition on LILRA3 expression. PBMCs from two donors were stimulated overnight with ssRNA40 together with neutralizing agents anakinra, certolizumab, tocilizumab and IFN-γ neutralizing antibodies and analysed for LILRA3 and IL6 expression by qPCR. Repeated measures ANOVA was used to calculate difference of ssRNA40+ neutralization to ssRNA40 alone and results displayed as mean ± SD
    Culture Pbmcs, supplied by Millipore, used in various techniques. Bioz Stars score: 89/100, based on 2549 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/culture pbmcs/product/Millipore
    Average 89 stars, based on 2549 article reviews
    Price from $9.99 to $1999.99
    culture pbmcs - by Bioz Stars, 2020-09
    89/100 stars
      Buy from Supplier

    85
    Millipore pbmc specimens against sd1 lps
    Fecal s-IgA antibody titers to <t>SD1-LPS</t> in group1 (cross symbol), group 2 (open square symbol), and group 3 (closed round symbol) monkeys, expressed as MN with MN ± SE, after challenge on day 00 and re-challenge on day 31 with SD11617 strain.
    Pbmc Specimens Against Sd1 Lps, supplied by Millipore, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pbmc specimens against sd1 lps/product/Millipore
    Average 85 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    pbmc specimens against sd1 lps - by Bioz Stars, 2020-09
    85/100 stars
      Buy from Supplier

    93
    Millipore human pbmcs
    Imatinib inhibits TNF-α production in human myeloid cells. Human <t>PBMCs,</t> monocytes, or macrophages were exposed to either saline or imatinib (1 h, 1 μM) followed by 3-h stimulation with 100 ng/ml <t>LPS.</t> ( A ) LPS-induced TNF-α levels in supernatants of all three cell types as determined by ELISA ( n = 5; * , P ≤ 0.01). ( B and C ) Imatinib dose-dependently inhibits LPS-induced TNF-α protein ( B ) ( n = 5; * , P ≤ 0.01) and TNF-α mRNA expression ( C ) ( n = 3; * , P ≤ 0.01) in human PBMCs.
    Human Pbmcs, supplied by Millipore, used in various techniques. Bioz Stars score: 93/100, based on 49 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human pbmcs/product/Millipore
    Average 93 stars, based on 49 article reviews
    Price from $9.99 to $1999.99
    human pbmcs - by Bioz Stars, 2020-09
    93/100 stars
      Buy from Supplier

    Image Search Results


    Features of ssRNA40-induced LILRA3 expression. a Kinetics of LILRA3 expression was compared to other ssRNA40 inducible cytokines. PBMCs from two donors were stimulated with ssRNA40 or ssRNA41 and the RNA was harvested at various time points. qPCR for LILRA3 , TNF , IFNb , IFNg , and IL1b expression, normalized to RPLP0 and relatively quantified to calibrator cDNA (pooled cDNA from 10. b Effect of cytokine inhibition on LILRA3 expression. PBMCs from two donors were stimulated overnight with ssRNA40 together with neutralizing agents anakinra, certolizumab, tocilizumab and IFN-γ neutralizing antibodies and analysed for LILRA3 and IL6 expression by qPCR. Repeated measures ANOVA was used to calculate difference of ssRNA40+ neutralization to ssRNA40 alone and results displayed as mean ± SD

    Journal: Retrovirology

    Article Title: TLR8 regulation of LILRA3 in monocytes is abrogated in human immunodeficiency virus infection and correlates to CD4 counts and virus loads

    doi: 10.1186/s12977-016-0248-y

    Figure Lengend Snippet: Features of ssRNA40-induced LILRA3 expression. a Kinetics of LILRA3 expression was compared to other ssRNA40 inducible cytokines. PBMCs from two donors were stimulated with ssRNA40 or ssRNA41 and the RNA was harvested at various time points. qPCR for LILRA3 , TNF , IFNb , IFNg , and IL1b expression, normalized to RPLP0 and relatively quantified to calibrator cDNA (pooled cDNA from 10. b Effect of cytokine inhibition on LILRA3 expression. PBMCs from two donors were stimulated overnight with ssRNA40 together with neutralizing agents anakinra, certolizumab, tocilizumab and IFN-γ neutralizing antibodies and analysed for LILRA3 and IL6 expression by qPCR. Repeated measures ANOVA was used to calculate difference of ssRNA40+ neutralization to ssRNA40 alone and results displayed as mean ± SD

    Article Snippet: PBMC and monocyte isolation and culture PBMCs were isolated using density gradient centrifugation by layering blood onto Biocoll (Millipore) and centrifugation at 1200×g for 20 min without brake.

    Techniques: Expressing, Real-time Polymerase Chain Reaction, Inhibition, Neutralization

    LILRA3 induction by TLRs. a Induction of LILRA3 and IL6 expression by a panel of TLR agonists. LILRA3 and IL6 expression was measured, using qPCR, as fold change to the unstimulated control from PBMCs stimulated for 24 h with Pam3CSK4 (P3C), heat killed Listeria monocytogenes (HKLM), polyinosinic-polycytidylic acid (Poly I:C) in high molecular weight (HMW) and low molecular weight (LMW) forms, lipopolysaccharide (LPS), flagellin, synthetic diacylated lipoprotein FSL-1, Imiquimod (R837), ssRNA40/LyoVec and CpG oligonucleotide ODN 2006. b Induction of LILRA3 by ssRNA40 on the transcriptional level. LILRA3 expression was measured as fold change to the unstimulated control from PBMCs stimulated for 24 h with uridine-rich HIV-derived ssRNA40, using ssRNA41 as control (n = 8). Wilcoxon matched-pairs signed rank test was used to compare the median between ssRNA41 and ssRNA40 stimulated LILRA3 expression. Scatter dot plot overlayed on bar graph displayed as median with interquartile range. c Induction of LILRA3 in PBMCS by ssRNA40 on the protein level. 30 × 10 6 /mL PBMCs from LILRA3 + / + and LILRA3 − / − donors were stimulated with 5 μg/mL ssRNA40. 40 µL from each supernatant was loaded and Ponceau S staining was used as loading control. The membrane was blotted for LILRA3 and IL6, using recombinant LILRA3 as protein control. d Expression of ssRNA40-induced LILRA3 by monocytes. PBMCs stimulated overnight with ssRNA40 and LPS were purified for CD14 + monocytes using magnetic-activated cell sorting. The resulting CD14 + positive and depleted fractions were analysed via qPCR for LILRA3 expression, normalized to RPLP0 and relatively quantified to calibrator cDNA (*p

    Journal: Retrovirology

    Article Title: TLR8 regulation of LILRA3 in monocytes is abrogated in human immunodeficiency virus infection and correlates to CD4 counts and virus loads

    doi: 10.1186/s12977-016-0248-y

    Figure Lengend Snippet: LILRA3 induction by TLRs. a Induction of LILRA3 and IL6 expression by a panel of TLR agonists. LILRA3 and IL6 expression was measured, using qPCR, as fold change to the unstimulated control from PBMCs stimulated for 24 h with Pam3CSK4 (P3C), heat killed Listeria monocytogenes (HKLM), polyinosinic-polycytidylic acid (Poly I:C) in high molecular weight (HMW) and low molecular weight (LMW) forms, lipopolysaccharide (LPS), flagellin, synthetic diacylated lipoprotein FSL-1, Imiquimod (R837), ssRNA40/LyoVec and CpG oligonucleotide ODN 2006. b Induction of LILRA3 by ssRNA40 on the transcriptional level. LILRA3 expression was measured as fold change to the unstimulated control from PBMCs stimulated for 24 h with uridine-rich HIV-derived ssRNA40, using ssRNA41 as control (n = 8). Wilcoxon matched-pairs signed rank test was used to compare the median between ssRNA41 and ssRNA40 stimulated LILRA3 expression. Scatter dot plot overlayed on bar graph displayed as median with interquartile range. c Induction of LILRA3 in PBMCS by ssRNA40 on the protein level. 30 × 10 6 /mL PBMCs from LILRA3 + / + and LILRA3 − / − donors were stimulated with 5 μg/mL ssRNA40. 40 µL from each supernatant was loaded and Ponceau S staining was used as loading control. The membrane was blotted for LILRA3 and IL6, using recombinant LILRA3 as protein control. d Expression of ssRNA40-induced LILRA3 by monocytes. PBMCs stimulated overnight with ssRNA40 and LPS were purified for CD14 + monocytes using magnetic-activated cell sorting. The resulting CD14 + positive and depleted fractions were analysed via qPCR for LILRA3 expression, normalized to RPLP0 and relatively quantified to calibrator cDNA (*p

    Article Snippet: PBMC and monocyte isolation and culture PBMCs were isolated using density gradient centrifugation by layering blood onto Biocoll (Millipore) and centrifugation at 1200×g for 20 min without brake.

    Techniques: Expressing, Real-time Polymerase Chain Reaction, Molecular Weight, Derivative Assay, Staining, Recombinant, Purification, FACS

    LILRA3 stimulation of cytokines and antigen presenting cells. a qPCR of LILRA3 induced gene expression of LILRA3 , IL - 6 , IL - 1A , IL - 1B and IL - 10 . Due to the different optimal concentrations of LILRA3 for various donors, the gene expression of all four donors were shown, with the corresponding mean ± SD. b Effect of LILRA3 on the antigen presentation mechanism of monocytes and B-cells. PBMCs were stimulated with varying concentrations of LILRA3 for 48 h and analysed by flow cytometry for expression of CD80, CD86, HLA-DR and HLA-ABC on monocytes (CD14 + CD33 + ) and B-cells (CD3 + CD19 + ). On monocytes, upregulation of HLA-ABC and CD80 was observed, whereas CD86 was downregulated. On B-cells, there was a slight but significant upregulation of HLA-DR and CD86. 1-way ANOVA with repeated measures Dunett post test was used to compare to 0 ng/mL LILRA3 control. Results expressed as mean ± SD from six donors. (*p

    Journal: Retrovirology

    Article Title: TLR8 regulation of LILRA3 in monocytes is abrogated in human immunodeficiency virus infection and correlates to CD4 counts and virus loads

    doi: 10.1186/s12977-016-0248-y

    Figure Lengend Snippet: LILRA3 stimulation of cytokines and antigen presenting cells. a qPCR of LILRA3 induced gene expression of LILRA3 , IL - 6 , IL - 1A , IL - 1B and IL - 10 . Due to the different optimal concentrations of LILRA3 for various donors, the gene expression of all four donors were shown, with the corresponding mean ± SD. b Effect of LILRA3 on the antigen presentation mechanism of monocytes and B-cells. PBMCs were stimulated with varying concentrations of LILRA3 for 48 h and analysed by flow cytometry for expression of CD80, CD86, HLA-DR and HLA-ABC on monocytes (CD14 + CD33 + ) and B-cells (CD3 + CD19 + ). On monocytes, upregulation of HLA-ABC and CD80 was observed, whereas CD86 was downregulated. On B-cells, there was a slight but significant upregulation of HLA-DR and CD86. 1-way ANOVA with repeated measures Dunett post test was used to compare to 0 ng/mL LILRA3 control. Results expressed as mean ± SD from six donors. (*p

    Article Snippet: PBMC and monocyte isolation and culture PBMCs were isolated using density gradient centrifugation by layering blood onto Biocoll (Millipore) and centrifugation at 1200×g for 20 min without brake.

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

    Validation of chemokine induction in TLR2 stimulated PBMCs. Human PBMCs were stimulated with TLR2 agonists for 16 h, and examined for CCL20 and CXCL6 expression by ELISA. PBMCs responded to both agonists that signal through TLR1/2 heterodimers (PAM 3 CSK 4 ) and TLR2/6 heterodimers (PAM 2 CSK 4 and DBS-2-217C). Means and standard deviations of triplicate samples are shown.

    Journal: Human Vaccines & Immunotherapeutics

    Article Title: Transcriptomal signatures of vaccine adjuvants and accessory immunostimulation of sentinel cells by toll-like receptor 2/6 agonists

    doi: 10.1080/21645515.2018.1480284

    Figure Lengend Snippet: Validation of chemokine induction in TLR2 stimulated PBMCs. Human PBMCs were stimulated with TLR2 agonists for 16 h, and examined for CCL20 and CXCL6 expression by ELISA. PBMCs responded to both agonists that signal through TLR1/2 heterodimers (PAM 3 CSK 4 ) and TLR2/6 heterodimers (PAM 2 CSK 4 and DBS-2-217C). Means and standard deviations of triplicate samples are shown.

    Article Snippet: Multiplexed cytokine analysis in PBMCs, SkMC, HMEC-1 and HFFs Cytokine and chemokine responses of PBMCs, SkMCs, HMEC-1, and HFFs were measured using methods previously reported by us , , with the following Milliplex kits: HCYTMAG-60K-PX41, HCYPMAG-63K, and HCYP2MAG-62K (EMD Millipore, Billerica MA).

    Techniques: Expressing, Enzyme-linked Immunosorbent Assay

    Chemotaxis of PBMCs toward TLR2/6- stimulated human foreskin fibroblasts. HFFs stimulated with TLR2/6, but not TLR1/2 agonists elicited functional chemotactic responses from lymphocytic populations in human PBMCs. Resting, adherent fibroblasts were stimulated with graded concentrations of agonists for 24 h in a chemotaxis plate. Chemotaxis of freshly isolated PBMCs was quantified by flow cytometry using lineage-specific antibodies. Means and SD on triplicate samples are shown.

    Journal: Human Vaccines & Immunotherapeutics

    Article Title: Transcriptomal signatures of vaccine adjuvants and accessory immunostimulation of sentinel cells by toll-like receptor 2/6 agonists

    doi: 10.1080/21645515.2018.1480284

    Figure Lengend Snippet: Chemotaxis of PBMCs toward TLR2/6- stimulated human foreskin fibroblasts. HFFs stimulated with TLR2/6, but not TLR1/2 agonists elicited functional chemotactic responses from lymphocytic populations in human PBMCs. Resting, adherent fibroblasts were stimulated with graded concentrations of agonists for 24 h in a chemotaxis plate. Chemotaxis of freshly isolated PBMCs was quantified by flow cytometry using lineage-specific antibodies. Means and SD on triplicate samples are shown.

    Article Snippet: Multiplexed cytokine analysis in PBMCs, SkMC, HMEC-1 and HFFs Cytokine and chemokine responses of PBMCs, SkMCs, HMEC-1, and HFFs were measured using methods previously reported by us , , with the following Milliplex kits: HCYTMAG-60K-PX41, HCYPMAG-63K, and HCYP2MAG-62K (EMD Millipore, Billerica MA).

    Techniques: Chemotaxis Assay, Functional Assay, Isolation, Flow Cytometry, Cytometry

    Fecal s-IgA antibody titers to SD1-LPS in group1 (cross symbol), group 2 (open square symbol), and group 3 (closed round symbol) monkeys, expressed as MN with MN ± SE, after challenge on day 00 and re-challenge on day 31 with SD11617 strain.

    Journal: Apmis

    Article Title: Evaluation of an intragastric challenge model for Shigella dysenteriae 1 in rhesus monkeys (Macaca mulatta) for the pre-clinical assessment of Shigella vaccine formulations

    doi: 10.1111/apm.12168

    Figure Lengend Snippet: Fecal s-IgA antibody titers to SD1-LPS in group1 (cross symbol), group 2 (open square symbol), and group 3 (closed round symbol) monkeys, expressed as MN with MN ± SE, after challenge on day 00 and re-challenge on day 31 with SD11617 strain.

    Article Snippet: Antibody-secreting cell assay Enzyme-linked immunospot (ELISPOT) assay was performed to enumerate IgA, IgG, and IgM ASCs on fresh PBMC specimens against SD1 LPS and SD1 Invaplex antigens using Multiscreen Immobilon-P filtration, 96-well plates, EMD Millipore Corporation, Billerica, MA, USA .

    Techniques:

    IgA, IgG, and IgM antibody titers against SD1-LPS in group 1 monkeys (cross symbol), group 2 (open square symbol), and group 3 (closed round symbol), expressed as geometric mean titer (GMT) with GMT±SE after challenge on day 00 and re-challenge on day 31 with SD11617 strain.

    Journal: Apmis

    Article Title: Evaluation of an intragastric challenge model for Shigella dysenteriae 1 in rhesus monkeys (Macaca mulatta) for the pre-clinical assessment of Shigella vaccine formulations

    doi: 10.1111/apm.12168

    Figure Lengend Snippet: IgA, IgG, and IgM antibody titers against SD1-LPS in group 1 monkeys (cross symbol), group 2 (open square symbol), and group 3 (closed round symbol), expressed as geometric mean titer (GMT) with GMT±SE after challenge on day 00 and re-challenge on day 31 with SD11617 strain.

    Article Snippet: Antibody-secreting cell assay Enzyme-linked immunospot (ELISPOT) assay was performed to enumerate IgA, IgG, and IgM ASCs on fresh PBMC specimens against SD1 LPS and SD1 Invaplex antigens using Multiscreen Immobilon-P filtration, 96-well plates, EMD Millipore Corporation, Billerica, MA, USA .

    Techniques:

    Imatinib inhibits TNF-α production in human myeloid cells. Human PBMCs, monocytes, or macrophages were exposed to either saline or imatinib (1 h, 1 μM) followed by 3-h stimulation with 100 ng/ml LPS. ( A ) LPS-induced TNF-α levels in supernatants of all three cell types as determined by ELISA ( n = 5; * , P ≤ 0.01). ( B and C ) Imatinib dose-dependently inhibits LPS-induced TNF-α protein ( B ) ( n = 5; * , P ≤ 0.01) and TNF-α mRNA expression ( C ) ( n = 3; * , P ≤ 0.01) in human PBMCs.

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

    Article Title: The kinase inhibitor imatinib mesylate inhibits TNF-? production in vitro and prevents TNF-dependent acute hepatic inflammation

    doi: 10.1073/pnas.0501758102

    Figure Lengend Snippet: Imatinib inhibits TNF-α production in human myeloid cells. Human PBMCs, monocytes, or macrophages were exposed to either saline or imatinib (1 h, 1 μM) followed by 3-h stimulation with 100 ng/ml LPS. ( A ) LPS-induced TNF-α levels in supernatants of all three cell types as determined by ELISA ( n = 5; * , P ≤ 0.01). ( B and C ) Imatinib dose-dependently inhibits LPS-induced TNF-α protein ( B ) ( n = 5; * , P ≤ 0.01) and TNF-α mRNA expression ( C ) ( n = 3; * , P ≤ 0.01) in human PBMCs.

    Article Snippet: Human PBMCs or CD14-selected monocytes were stimulated with LPS at a concentration of 100 ng/ml for 12 h in the presence of either solvent or the indicated kinase inhibitor ( , SB203580, PD98059, and JNK-Inhibitor II, all used in a final concentration of 35 μM and purchased from Calbiochem).

    Techniques: Enzyme-linked Immunosorbent Assay, Expressing