human peripheral blood mononuclear cells pbmcs Search Results


  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • N/A
    High quality high viability 90 peripheral blood mononuclear cells PBMCs from patients with non small cell lung carcinoma NSCLC sourced from a world renowned blood center Used for a wide
      Buy from Supplier

    N/A
    Cryopreserved Human Peripheral Blood Mononuclear Cells are available as positive and negative controls for T cell monitoring in ELISPOT ELISA cytokine bead array tetramer pentamer and flow cytometry assays A
      Buy from Supplier

    99
    Millipore histopaque 1077
    Histopaque 1077, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 12612 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/histopaque 1077/product/Millipore
    Average 99 stars, based on 12612 article reviews
    Price from $9.99 to $1999.99
    histopaque 1077 - by Bioz Stars, 2021-01
    99/100 stars
      Buy from Supplier

    96
    Millipore ceramide
    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
    https://www.bioz.com/result/ceramide/product/Millipore
    Average 96 stars, based on 483 article reviews
    Price from $9.99 to $1999.99
    ceramide - by Bioz Stars, 2021-01
    96/100 stars
      Buy from Supplier

    pbmcs  (lonza)
    92
    lonza pbmcs
    Pbmcs, supplied by lonza, used in various techniques. Bioz Stars score: 92/100, based on 1855 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pbmcs/product/lonza
    Average 92 stars, based on 1855 article reviews
    Price from $9.99 to $1999.99
    pbmcs - by Bioz Stars, 2021-01
    92/100 stars
      Buy from Supplier

    90
    ATCC primary peripheral blood mononuclear cells pbmc normal human
    Knockdown of thymidylate synthase (TS) induces programmed death-ligand 1 (PD-L1) expression in non-small-cell lung cancer (NSCLC) cells and decreases the production of interleukin-2 (IL-2) by activated T cells in the NSCLC and T cell coculture system. (A–D), CL1-5 or CL141 cells transfected with control-siRNA (siCtrl) or TS -siRNA (siTS) oligonucleotides were lysed and analyzed by qRT-PCR (A), flow cytometry (B), or immunoblotting (C, D) 72 hours after transfection. Data are shown as means and SD for three independent experiments (n=3). (E, F) CL1-5 or CL141 cells transfected with control-siRNA (siCtrl), TS-siRNA (siTS) and RelA-siRNA (siRelA) oligonucleotides in different combination were lysed and analyzed by qRT-PCR (E) or immunoblotting (F), 72 hours after transfection. (G, H) CL1-5 or CL141 cells were transfected with siCtrl or siTS siRNA oligonucleotides for 24 hours and followed by cocultured with Jurkat T-cells or <t>PBMCs</t> at different cancer to T cell ratios in the presence of the 1×T cell stimulation cocktail for additional 48 hours. (G) IL-2 levels were measured by ELISA. (H) The levels of CD69 and intracellular IL-2 produced by Jurkat T-cells or PBMCs were measured by flow cytometry. **P
    Primary Peripheral Blood Mononuclear Cells Pbmc Normal Human, supplied by ATCC, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/primary peripheral blood mononuclear cells pbmc normal human/product/ATCC
    Average 90 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    primary peripheral blood mononuclear cells pbmc normal human - by Bioz Stars, 2021-01
    90/100 stars
      Buy from Supplier

    92
    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: 92/100, based on 112 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human peripheral blood mononuclear cells pbmcs/product/Axis-Shield Diagnostics
    Average 92 stars, based on 112 article reviews
    Price from $9.99 to $1999.99
    human peripheral blood mononuclear cells pbmcs - by Bioz Stars, 2021-01
    92/100 stars
      Buy from Supplier

    92
    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
    https://www.bioz.com/result/human pbmcs/product/AllCells LLC
    Average 92 stars, based on 50 article reviews
    Price from $9.99 to $1999.99
    human pbmcs - by Bioz Stars, 2021-01
    92/100 stars
      Buy from Supplier

    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
    https://www.bioz.com/result/human pbmcs/product/Axis-Shield Diagnostics
    Average 92 stars, based on 140 article reviews
    Price from $9.99 to $1999.99
    human pbmcs - by Bioz Stars, 2021-01
    92/100 stars
      Buy from Supplier

    97
    Millipore peripheral blood mononuclear cells 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:
    Peripheral Blood Mononuclear Cells Pbmcs, supplied by Millipore, used in various techniques. Bioz Stars score: 97/100, based on 3440 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/peripheral blood mononuclear cells pbmcs/product/Millipore
    Average 97 stars, based on 3440 article reviews
    Price from $9.99 to $1999.99
    peripheral blood mononuclear cells pbmcs - by Bioz Stars, 2021-01
    97/100 stars
      Buy from Supplier

    92
    Becton Dickinson human pbmcs
    IL-22 + upregulation occurs in <t>CD4</t> + 8 - T cells that are IL-17 negative but express IFNγ. (A) Cytokine production for live CD3 + CD4 + CD8 − T cells from human <t>PBMC</t> cultures that were stimulated with anti-CD3 and anti-CD28 antibodies and allogeneic APCs for six days with DMSO or 50 µM 3-HAA. Panel (right) depicts fold-changes in 9 donors relative to DMSO. *, p
    Human Pbmcs, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 92/100, based on 603 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human pbmcs/product/Becton Dickinson
    Average 92 stars, based on 603 article reviews
    Price from $9.99 to $1999.99
    human pbmcs - by Bioz Stars, 2021-01
    92/100 stars
      Buy from Supplier

    92
    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: 92/100, based on 1342 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/peripheral blood mononuclear cells pbmcs/product/Axis-Shield Diagnostics
    Average 92 stars, based on 1342 article reviews
    Price from $9.99 to $1999.99
    peripheral blood mononuclear cells pbmcs - by Bioz Stars, 2021-01
    92/100 stars
      Buy from Supplier

    92
    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: 92/100, based on 109 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human peripheral blood mononuclear cells pbmcs/product/Biochrom
    Average 92 stars, based on 109 article reviews
    Price from $9.99 to $1999.99
    human peripheral blood mononuclear cells pbmcs - by Bioz Stars, 2021-01
    92/100 stars
      Buy from Supplier

    92
    Thermo Fisher 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.
    Pbmcs, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 31889 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pbmcs/product/Thermo Fisher
    Average 92 stars, based on 31889 article reviews
    Price from $9.99 to $1999.99
    pbmcs - by Bioz Stars, 2021-01
    92/100 stars
      Buy from Supplier

    91
    ActivX human pbmcs
    Lu AF58786 inhibits phosphorylation of LRRK2, Rab10 and Rab12 in cultured and immune stimulated human <t>PBMCs.</t> ( A ) LRRK2 (pSer910, <t>pSer935,</t> pSer955 and pSer973) phospho-site hits plotted with mean and individual replicate values of the quantified log 10 -transformed CMPD/DMSO ratio as well as the associated q-value of the performed mean rank (n = 5–8). ( B ) Rab10 (pThr73) and Rab12 (pSer106) switch II domain phospho-site hits plotted with mean and individual replicate values of the quantified log 10 -transformed CMPD/DMSO ratio as well as the associated q-value of the performed mean rank (n = 5–8). CMPD, compound (Lu AF58786). DMSO, dimethyl sulfoxide.
    Human Pbmcs, supplied by ActivX, used in various techniques. Bioz Stars score: 91/100, based on 37 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human pbmcs/product/ActivX
    Average 91 stars, based on 37 article reviews
    Price from $9.99 to $1999.99
    human pbmcs - by Bioz Stars, 2021-01
    91/100 stars
      Buy from Supplier

    92
    Biochrom peripheral blood mononuclear cells pbmc
    miR-20b expression in T cells and in <t>PBMC</t> from natalizumab-associated progressive multifocal leukoencephalopathy (PML). (A) PBMC were isolated using a <t>Ficoll</t> gradient and CD4+ T cells were sorted with magnetic beads ( n = 9; randomly selected patients of our initial longitudinal cohort consisting of 17 patients). miR-20b expression (qPCR) was measured in the CD4+ and CD4-negative fraction and compared using t- test. (B) CD4+ cells were activated by plate-bound anti-CD3-antibody and soluble anti-CD28-antibody without further cytokines, termed “Th0”, and with IL-1 (10 ng/mL), IL-6 (10 ng/mL), IL-23 (10 ng/mL) and TGF- β (5 ng/mL), termed “Th17”, as well as freshly isolated CD4+ T cells, termed “fresh”. miR-20b levels were measured by qPCR. (C) miR-20b levels in PBMC from natalizumab-treated patients ( n = 9, left bar) were compared to miR-20b levels in two PBMC samples from patients with natalizumab-associated PML (each patient one bar).
    Peripheral Blood Mononuclear Cells Pbmc, supplied by Biochrom, used in various techniques. Bioz Stars score: 92/100, based on 447 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/peripheral blood mononuclear cells pbmc/product/Biochrom
    Average 92 stars, based on 447 article reviews
    Price from $9.99 to $1999.99
    peripheral blood mononuclear cells pbmc - by Bioz Stars, 2021-01
    92/100 stars
      Buy from Supplier

    92
    Lonza human pbmc human peripheral blood mononuclear cells cryopreserved
    miR-20b expression in T cells and in <t>PBMC</t> from natalizumab-associated progressive multifocal leukoencephalopathy (PML). (A) PBMC were isolated using a <t>Ficoll</t> gradient and CD4+ T cells were sorted with magnetic beads ( n = 9; randomly selected patients of our initial longitudinal cohort consisting of 17 patients). miR-20b expression (qPCR) was measured in the CD4+ and CD4-negative fraction and compared using t- test. (B) CD4+ cells were activated by plate-bound anti-CD3-antibody and soluble anti-CD28-antibody without further cytokines, termed “Th0”, and with IL-1 (10 ng/mL), IL-6 (10 ng/mL), IL-23 (10 ng/mL) and TGF- β (5 ng/mL), termed “Th17”, as well as freshly isolated CD4+ T cells, termed “fresh”. miR-20b levels were measured by qPCR. (C) miR-20b levels in PBMC from natalizumab-treated patients ( n = 9, left bar) were compared to miR-20b levels in two PBMC samples from patients with natalizumab-associated PML (each patient one bar).
    Human Pbmc Human Peripheral Blood Mononuclear Cells Cryopreserved, supplied by Lonza, used in various techniques. Bioz Stars score: 92/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human pbmc human peripheral blood mononuclear cells cryopreserved/product/Lonza
    Average 92 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    human pbmc human peripheral blood mononuclear cells cryopreserved - by Bioz Stars, 2021-01
    92/100 stars
      Buy from Supplier

    92
    Nycomed human peripheral blood mononuclear cells pbmcs
    miR-20b expression in T cells and in <t>PBMC</t> from natalizumab-associated progressive multifocal leukoencephalopathy (PML). (A) PBMC were isolated using a <t>Ficoll</t> gradient and CD4+ T cells were sorted with magnetic beads ( n = 9; randomly selected patients of our initial longitudinal cohort consisting of 17 patients). miR-20b expression (qPCR) was measured in the CD4+ and CD4-negative fraction and compared using t- test. (B) CD4+ cells were activated by plate-bound anti-CD3-antibody and soluble anti-CD28-antibody without further cytokines, termed “Th0”, and with IL-1 (10 ng/mL), IL-6 (10 ng/mL), IL-23 (10 ng/mL) and TGF- β (5 ng/mL), termed “Th17”, as well as freshly isolated CD4+ T cells, termed “fresh”. miR-20b levels were measured by qPCR. (C) miR-20b levels in PBMC from natalizumab-treated patients ( n = 9, left bar) were compared to miR-20b levels in two PBMC samples from patients with natalizumab-associated PML (each patient one bar).
    Human Peripheral Blood Mononuclear Cells Pbmcs, supplied by Nycomed, used in various techniques. Bioz Stars score: 92/100, based on 35 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human peripheral blood mononuclear cells pbmcs/product/Nycomed
    Average 92 stars, based on 35 article reviews
    Price from $9.99 to $1999.99
    human peripheral blood mononuclear cells pbmcs - by Bioz Stars, 2021-01
    92/100 stars
      Buy from Supplier

    92
    AllCells LLC pbmcs
    HIV-1 infection in humanized mice passively infused with b12-IgA or b12-IgG isotype. (A) Neutralization activity of anti-HIV human mAb b12-IgA. Reporter cell line TZM-bl cells that express CD4, CXCR4, CCR5, and a Tat-responsive reporter gene for luciferase were infected with 200 TCID 50 of replication-defective pseudovirus containing Env (SF162.LS) in the presence of various concentrations of anti-HIV monoclonal antibodies. Neutralization activity was measured by the reduction in luciferase reporter gene expression after a single round of pseudovirus infection in TZM-bl cells in triplicate. b12-IgG1 and the recombinant b12-IgA2 were compared at indicated concentrations. (B) Antibody level in circulation after passive transfer. <t>NSG-hu</t> mice were injected intravenously with various concentrations of purified b12-IgA2 (pIgA: mIgA = 1:1, mass ratio) or b12-IgG1 and the blood was collected after 4 hours. The plasma antibody concentrations were measured using ELISA. (n = 4-6, mean ± SEM). (C-D) Concentrations of b12 antibodies in plasma (C) or genital secretions (D) at the time of challenge. NSG-hu mice were injected intravenously with either 200 μg of b12-IgA2 or 20 μg of b12-IgG1 per mouse. Blood and genital secretions were collected after 4 hours. (E-I) Peripheral CD4 + T cell loss after HIV-1 challenge in NSG-hu mice injected with different b12 antibody isotypes; (E) purified human IgG/κ control antibody (hIgG/κ); (F) b12-IgA2 includes both monomeric and polymeric IgA as described in (B), (b12IgA2 [M+P]); (G) b12-IgG1; (H) b12-IgA2 monomer only (b12IgA2 [M]). Mice were challenged intravaginally with HIV-1 JR-CSF . (I) Average percent CD4 + T cells in CD3 T cells in <t>PBMCs</t> at each time points (n = 5-9). Statistical analysis was performed using unpaired t test. (* P = .0313 and *** P
    Pbmcs, supplied by AllCells LLC, used in various techniques. Bioz Stars score: 92/100, based on 153 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pbmcs/product/AllCells LLC
    Average 92 stars, based on 153 article reviews
    Price from $9.99 to $1999.99
    pbmcs - by Bioz Stars, 2021-01
    92/100 stars
      Buy from Supplier

    90
    ATCC human peripheral blood mononuclear cells pbmcs
    HIV-1 infection in humanized mice passively infused with b12-IgA or b12-IgG isotype. (A) Neutralization activity of anti-HIV human mAb b12-IgA. Reporter cell line TZM-bl cells that express CD4, CXCR4, CCR5, and a Tat-responsive reporter gene for luciferase were infected with 200 TCID 50 of replication-defective pseudovirus containing Env (SF162.LS) in the presence of various concentrations of anti-HIV monoclonal antibodies. Neutralization activity was measured by the reduction in luciferase reporter gene expression after a single round of pseudovirus infection in TZM-bl cells in triplicate. b12-IgG1 and the recombinant b12-IgA2 were compared at indicated concentrations. (B) Antibody level in circulation after passive transfer. <t>NSG-hu</t> mice were injected intravenously with various concentrations of purified b12-IgA2 (pIgA: mIgA = 1:1, mass ratio) or b12-IgG1 and the blood was collected after 4 hours. The plasma antibody concentrations were measured using ELISA. (n = 4-6, mean ± SEM). (C-D) Concentrations of b12 antibodies in plasma (C) or genital secretions (D) at the time of challenge. NSG-hu mice were injected intravenously with either 200 μg of b12-IgA2 or 20 μg of b12-IgG1 per mouse. Blood and genital secretions were collected after 4 hours. (E-I) Peripheral CD4 + T cell loss after HIV-1 challenge in NSG-hu mice injected with different b12 antibody isotypes; (E) purified human IgG/κ control antibody (hIgG/κ); (F) b12-IgA2 includes both monomeric and polymeric IgA as described in (B), (b12IgA2 [M+P]); (G) b12-IgG1; (H) b12-IgA2 monomer only (b12IgA2 [M]). Mice were challenged intravaginally with HIV-1 JR-CSF . (I) Average percent CD4 + T cells in CD3 T cells in <t>PBMCs</t> at each time points (n = 5-9). Statistical analysis was performed using unpaired t test. (* P = .0313 and *** P
    Human Peripheral Blood Mononuclear Cells Pbmcs, supplied by ATCC, used in various techniques. Bioz Stars score: 90/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human peripheral blood mononuclear cells pbmcs/product/ATCC
    Average 90 stars, based on 15 article reviews
    Price from $9.99 to $1999.99
    human peripheral blood mononuclear cells pbmcs - by Bioz Stars, 2021-01
    90/100 stars
      Buy from Supplier

    92
    Becton Dickinson peripheral blood mononuclear cells pbmcs
    HIV-1 infection in humanized mice passively infused with b12-IgA or b12-IgG isotype. (A) Neutralization activity of anti-HIV human mAb b12-IgA. Reporter cell line TZM-bl cells that express CD4, CXCR4, CCR5, and a Tat-responsive reporter gene for luciferase were infected with 200 TCID 50 of replication-defective pseudovirus containing Env (SF162.LS) in the presence of various concentrations of anti-HIV monoclonal antibodies. Neutralization activity was measured by the reduction in luciferase reporter gene expression after a single round of pseudovirus infection in TZM-bl cells in triplicate. b12-IgG1 and the recombinant b12-IgA2 were compared at indicated concentrations. (B) Antibody level in circulation after passive transfer. <t>NSG-hu</t> mice were injected intravenously with various concentrations of purified b12-IgA2 (pIgA: mIgA = 1:1, mass ratio) or b12-IgG1 and the blood was collected after 4 hours. The plasma antibody concentrations were measured using ELISA. (n = 4-6, mean ± SEM). (C-D) Concentrations of b12 antibodies in plasma (C) or genital secretions (D) at the time of challenge. NSG-hu mice were injected intravenously with either 200 μg of b12-IgA2 or 20 μg of b12-IgG1 per mouse. Blood and genital secretions were collected after 4 hours. (E-I) Peripheral CD4 + T cell loss after HIV-1 challenge in NSG-hu mice injected with different b12 antibody isotypes; (E) purified human IgG/κ control antibody (hIgG/κ); (F) b12-IgA2 includes both monomeric and polymeric IgA as described in (B), (b12IgA2 [M+P]); (G) b12-IgG1; (H) b12-IgA2 monomer only (b12IgA2 [M]). Mice were challenged intravaginally with HIV-1 JR-CSF . (I) Average percent CD4 + T cells in CD3 T cells in <t>PBMCs</t> at each time points (n = 5-9). Statistical analysis was performed using unpaired t test. (* P = .0313 and *** P
    Peripheral Blood Mononuclear Cells Pbmcs, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 92/100, based on 950 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/peripheral blood mononuclear cells pbmcs/product/Becton Dickinson
    Average 92 stars, based on 950 article reviews
    Price from $9.99 to $1999.99
    peripheral blood mononuclear cells pbmcs - by Bioz Stars, 2021-01
    92/100 stars
      Buy from Supplier

    92
    Nycomed peripheral blood mononuclear cells pbmcs
    HIV-1 infection in humanized mice passively infused with b12-IgA or b12-IgG isotype. (A) Neutralization activity of anti-HIV human mAb b12-IgA. Reporter cell line TZM-bl cells that express CD4, CXCR4, CCR5, and a Tat-responsive reporter gene for luciferase were infected with 200 TCID 50 of replication-defective pseudovirus containing Env (SF162.LS) in the presence of various concentrations of anti-HIV monoclonal antibodies. Neutralization activity was measured by the reduction in luciferase reporter gene expression after a single round of pseudovirus infection in TZM-bl cells in triplicate. b12-IgG1 and the recombinant b12-IgA2 were compared at indicated concentrations. (B) Antibody level in circulation after passive transfer. <t>NSG-hu</t> mice were injected intravenously with various concentrations of purified b12-IgA2 (pIgA: mIgA = 1:1, mass ratio) or b12-IgG1 and the blood was collected after 4 hours. The plasma antibody concentrations were measured using ELISA. (n = 4-6, mean ± SEM). (C-D) Concentrations of b12 antibodies in plasma (C) or genital secretions (D) at the time of challenge. NSG-hu mice were injected intravenously with either 200 μg of b12-IgA2 or 20 μg of b12-IgG1 per mouse. Blood and genital secretions were collected after 4 hours. (E-I) Peripheral CD4 + T cell loss after HIV-1 challenge in NSG-hu mice injected with different b12 antibody isotypes; (E) purified human IgG/κ control antibody (hIgG/κ); (F) b12-IgA2 includes both monomeric and polymeric IgA as described in (B), (b12IgA2 [M+P]); (G) b12-IgG1; (H) b12-IgA2 monomer only (b12IgA2 [M]). Mice were challenged intravaginally with HIV-1 JR-CSF . (I) Average percent CD4 + T cells in CD3 T cells in <t>PBMCs</t> at each time points (n = 5-9). Statistical analysis was performed using unpaired t test. (* P = .0313 and *** P
    Peripheral Blood Mononuclear Cells Pbmcs, supplied by Nycomed, used in various techniques. Bioz Stars score: 92/100, based on 375 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/peripheral blood mononuclear cells pbmcs/product/Nycomed
    Average 92 stars, based on 375 article reviews
    Price from $9.99 to $1999.99
    peripheral blood mononuclear cells pbmcs - by Bioz Stars, 2021-01
    92/100 stars
      Buy from Supplier

    92
    Valiant peripheral blood mononuclear cells pbmcs
    HIV-1 infection in humanized mice passively infused with b12-IgA or b12-IgG isotype. (A) Neutralization activity of anti-HIV human mAb b12-IgA. Reporter cell line TZM-bl cells that express CD4, CXCR4, CCR5, and a Tat-responsive reporter gene for luciferase were infected with 200 TCID 50 of replication-defective pseudovirus containing Env (SF162.LS) in the presence of various concentrations of anti-HIV monoclonal antibodies. Neutralization activity was measured by the reduction in luciferase reporter gene expression after a single round of pseudovirus infection in TZM-bl cells in triplicate. b12-IgG1 and the recombinant b12-IgA2 were compared at indicated concentrations. (B) Antibody level in circulation after passive transfer. <t>NSG-hu</t> mice were injected intravenously with various concentrations of purified b12-IgA2 (pIgA: mIgA = 1:1, mass ratio) or b12-IgG1 and the blood was collected after 4 hours. The plasma antibody concentrations were measured using ELISA. (n = 4-6, mean ± SEM). (C-D) Concentrations of b12 antibodies in plasma (C) or genital secretions (D) at the time of challenge. NSG-hu mice were injected intravenously with either 200 μg of b12-IgA2 or 20 μg of b12-IgG1 per mouse. Blood and genital secretions were collected after 4 hours. (E-I) Peripheral CD4 + T cell loss after HIV-1 challenge in NSG-hu mice injected with different b12 antibody isotypes; (E) purified human IgG/κ control antibody (hIgG/κ); (F) b12-IgA2 includes both monomeric and polymeric IgA as described in (B), (b12IgA2 [M+P]); (G) b12-IgG1; (H) b12-IgA2 monomer only (b12IgA2 [M]). Mice were challenged intravaginally with HIV-1 JR-CSF . (I) Average percent CD4 + T cells in CD3 T cells in <t>PBMCs</t> at each time points (n = 5-9). Statistical analysis was performed using unpaired t test. (* P = .0313 and *** P
    Peripheral Blood Mononuclear Cells Pbmcs, supplied by Valiant, used in various techniques. Bioz Stars score: 92/100, based on 193 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/peripheral blood mononuclear cells pbmcs/product/Valiant
    Average 92 stars, based on 193 article reviews
    Price from $9.99 to $1999.99
    peripheral blood mononuclear cells pbmcs - by Bioz Stars, 2021-01
    92/100 stars
      Buy from Supplier

    90
    10X Genomics human pbmc cite seq dataset
    The performance of BREM-SC with the public human <t>PBMC</t> <t>CITE-Seq</t> dataset (from 10X Genomics). The UMAP projection of cells are colored by the ground truth ( 4A ) and BREM-SC clustering results ( 4B ).
    Human Pbmc Cite Seq Dataset, supplied by 10X Genomics, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human pbmc cite seq dataset/product/10X Genomics
    Average 90 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    human pbmc cite seq dataset - by Bioz Stars, 2021-01
    90/100 stars
      Buy from Supplier

    N/A
    Human Peripheral Blood Mononuclear Cells are available as positive and negative controls for T cell monitoring in ELISPOT ELISA cytokine bead array tetramer pentamer and flow cytometry assays A peripheral
      Buy from Supplier

    Image Search Results


    Knockdown of thymidylate synthase (TS) induces programmed death-ligand 1 (PD-L1) expression in non-small-cell lung cancer (NSCLC) cells and decreases the production of interleukin-2 (IL-2) by activated T cells in the NSCLC and T cell coculture system. (A–D), CL1-5 or CL141 cells transfected with control-siRNA (siCtrl) or TS -siRNA (siTS) oligonucleotides were lysed and analyzed by qRT-PCR (A), flow cytometry (B), or immunoblotting (C, D) 72 hours after transfection. Data are shown as means and SD for three independent experiments (n=3). (E, F) CL1-5 or CL141 cells transfected with control-siRNA (siCtrl), TS-siRNA (siTS) and RelA-siRNA (siRelA) oligonucleotides in different combination were lysed and analyzed by qRT-PCR (E) or immunoblotting (F), 72 hours after transfection. (G, H) CL1-5 or CL141 cells were transfected with siCtrl or siTS siRNA oligonucleotides for 24 hours and followed by cocultured with Jurkat T-cells or PBMCs at different cancer to T cell ratios in the presence of the 1×T cell stimulation cocktail for additional 48 hours. (G) IL-2 levels were measured by ELISA. (H) The levels of CD69 and intracellular IL-2 produced by Jurkat T-cells or PBMCs were measured by flow cytometry. **P

    Journal: Journal for Immunotherapy of Cancer

    Article Title: Antimetabolite pemetrexed primes a favorable tumor microenvironment for immune checkpoint blockade therapy

    doi: 10.1136/jitc-2020-001392

    Figure Lengend Snippet: Knockdown of thymidylate synthase (TS) induces programmed death-ligand 1 (PD-L1) expression in non-small-cell lung cancer (NSCLC) cells and decreases the production of interleukin-2 (IL-2) by activated T cells in the NSCLC and T cell coculture system. (A–D), CL1-5 or CL141 cells transfected with control-siRNA (siCtrl) or TS -siRNA (siTS) oligonucleotides were lysed and analyzed by qRT-PCR (A), flow cytometry (B), or immunoblotting (C, D) 72 hours after transfection. Data are shown as means and SD for three independent experiments (n=3). (E, F) CL1-5 or CL141 cells transfected with control-siRNA (siCtrl), TS-siRNA (siTS) and RelA-siRNA (siRelA) oligonucleotides in different combination were lysed and analyzed by qRT-PCR (E) or immunoblotting (F), 72 hours after transfection. (G, H) CL1-5 or CL141 cells were transfected with siCtrl or siTS siRNA oligonucleotides for 24 hours and followed by cocultured with Jurkat T-cells or PBMCs at different cancer to T cell ratios in the presence of the 1×T cell stimulation cocktail for additional 48 hours. (G) IL-2 levels were measured by ELISA. (H) The levels of CD69 and intracellular IL-2 produced by Jurkat T-cells or PBMCs were measured by flow cytometry. **P

    Article Snippet: Human peripheral blood mononuclear cells (PBMCs) were isolated from healthy blood donors.

    Techniques: Expressing, Transfection, Quantitative RT-PCR, Flow Cytometry, Cell Stimulation, Enzyme-linked Immunosorbent Assay, Produced

    Pemetrexed (PEM) and 5-fluorouracil (5-FU) suppress the production of interleukin-2 (IL-2) and interferon (IFN)-γ by activated T cells in the non-small-cell lung cancer (NSCLC) and T cell coculture system. (A–D) CL1-5 or CL141 cells were preincubated with 100 nM PEM, 5 µM 5-FU or the vehicle control (PBS) for 48 hours, and subsequently cocultured with activated Jurkat T-cells (A, B) or PBMCs (C, D) at different cancer to T cell ratios in the presence of 1× T cell stimulation cocktail for additional 48 hours. The levels of IL-2 and IFN-γ were measured by ELISA. Data are shown as means and SD for three independent experiments (n=3). (E–G) CL1-5 or CL141 cells were preincubated with 100 nM PEM, 5 µM 5-FU or the vehicle control (PBS) for 48 hours, and subsequently cocultured with activated Jurkat T-cells in the presence of 1× T cell stimulation cocktail for additional 48 hours. The levels of CD69 and intracecllular IL-2 produced by Jurkat T-cells were measured by flow cytometry. (E) Representative dot plots for the indicated cell percentages determined by flow cytometry. (F, G) Quantitative plots for the CD69 and intracellular IL-2 staining in CD45 + T-cells. (H) T-cell-meditated killing of PD-L1-expressing CL141 NSCLC cells. CL141 cells stably expressing nuclear RFP protein were pretreated with or without 50 nM PEM for 48 hours and cocultured with activated Jurkat T-cells with or without 10 µg/mL of anti-PD-L1 antibody for additional 48 hours. Relative cell viability was measured by RFP signaling after 48 hours of coincubation and the results were normalized at the zero-time point. Data are shown as means and s.e.m. for three independent experiments (n=3). *P

    Journal: Journal for Immunotherapy of Cancer

    Article Title: Antimetabolite pemetrexed primes a favorable tumor microenvironment for immune checkpoint blockade therapy

    doi: 10.1136/jitc-2020-001392

    Figure Lengend Snippet: Pemetrexed (PEM) and 5-fluorouracil (5-FU) suppress the production of interleukin-2 (IL-2) and interferon (IFN)-γ by activated T cells in the non-small-cell lung cancer (NSCLC) and T cell coculture system. (A–D) CL1-5 or CL141 cells were preincubated with 100 nM PEM, 5 µM 5-FU or the vehicle control (PBS) for 48 hours, and subsequently cocultured with activated Jurkat T-cells (A, B) or PBMCs (C, D) at different cancer to T cell ratios in the presence of 1× T cell stimulation cocktail for additional 48 hours. The levels of IL-2 and IFN-γ were measured by ELISA. Data are shown as means and SD for three independent experiments (n=3). (E–G) CL1-5 or CL141 cells were preincubated with 100 nM PEM, 5 µM 5-FU or the vehicle control (PBS) for 48 hours, and subsequently cocultured with activated Jurkat T-cells in the presence of 1× T cell stimulation cocktail for additional 48 hours. The levels of CD69 and intracecllular IL-2 produced by Jurkat T-cells were measured by flow cytometry. (E) Representative dot plots for the indicated cell percentages determined by flow cytometry. (F, G) Quantitative plots for the CD69 and intracellular IL-2 staining in CD45 + T-cells. (H) T-cell-meditated killing of PD-L1-expressing CL141 NSCLC cells. CL141 cells stably expressing nuclear RFP protein were pretreated with or without 50 nM PEM for 48 hours and cocultured with activated Jurkat T-cells with or without 10 µg/mL of anti-PD-L1 antibody for additional 48 hours. Relative cell viability was measured by RFP signaling after 48 hours of coincubation and the results were normalized at the zero-time point. Data are shown as means and s.e.m. for three independent experiments (n=3). *P

    Article Snippet: Human peripheral blood mononuclear cells (PBMCs) were isolated from healthy blood donors.

    Techniques: Cell Stimulation, Enzyme-linked Immunosorbent Assay, Produced, Flow Cytometry, Staining, Expressing, Stable Transfection

    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

    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

    IL-22 + upregulation occurs in CD4 + 8 - T cells that are IL-17 negative but express IFNγ. (A) Cytokine production for live CD3 + CD4 + CD8 − T cells from human PBMC cultures that were stimulated with anti-CD3 and anti-CD28 antibodies and allogeneic APCs for six days with DMSO or 50 µM 3-HAA. Panel (right) depicts fold-changes in 9 donors relative to DMSO. *, p

    Journal: PLoS ONE

    Article Title: Identification of Cinnabarinic Acid as a Novel Endogenous Aryl Hydrocarbon Receptor Ligand That Drives IL-22 Production

    doi: 10.1371/journal.pone.0087877

    Figure Lengend Snippet: IL-22 + upregulation occurs in CD4 + 8 - T cells that are IL-17 negative but express IFNγ. (A) Cytokine production for live CD3 + CD4 + CD8 − T cells from human PBMC cultures that were stimulated with anti-CD3 and anti-CD28 antibodies and allogeneic APCs for six days with DMSO or 50 µM 3-HAA. Panel (right) depicts fold-changes in 9 donors relative to DMSO. *, p

    Article Snippet: For naïve CD4+ T cell sorting, human PBMCs from adult donors or from de-identified cord blood (MD Anderson) were stained with anti-CD3-Alexa700 (BD), anti-CD4-ECD (Invitrogen), anti-CD8-PeCy5.5 (Invitrogen), anti-CD45RA-FITC (BD), anti-CD95-APC (BD), anti-CD25-PE (BD), anti-CCR7-PeCy7 (BD), and anti-CD27-APCCy7 (eBioscience).

    Techniques:

    CA is an endogenous AHR agonist generated by oxidative dimerization of 3-HAA. (A) Metabolic pathways downstream of 3-HAA catalyzed by the enzyme, 3-hydroxyanthranilate 3,4-dioxygenase (HAAO), including intermediates upstream of PA and QA. (B) Flow cytometric analysis of CD4 + T cells in human PBMCs stimulated in the presence of varying concentrations of 3-HAA (50 µM or 100 µM) with or without the HAAO inhibitor, 4-F-3-HAA (50 µM). Data are representative of three experiments. (C) An alternative pathway of 3-HAA metabolism can generate CA either by non-enzymatic (oxidation) or enzymatic (such as laccase or ceruloplasmin) processes. (D) Fluorescence [measured in relative fluorescence units (RFUs)] of the AHR-responsive reporter construct in cells incubated with varying concentrations of 3-HAA or CA. Tryptamine was a positive control. *, p

    Journal: PLoS ONE

    Article Title: Identification of Cinnabarinic Acid as a Novel Endogenous Aryl Hydrocarbon Receptor Ligand That Drives IL-22 Production

    doi: 10.1371/journal.pone.0087877

    Figure Lengend Snippet: CA is an endogenous AHR agonist generated by oxidative dimerization of 3-HAA. (A) Metabolic pathways downstream of 3-HAA catalyzed by the enzyme, 3-hydroxyanthranilate 3,4-dioxygenase (HAAO), including intermediates upstream of PA and QA. (B) Flow cytometric analysis of CD4 + T cells in human PBMCs stimulated in the presence of varying concentrations of 3-HAA (50 µM or 100 µM) with or without the HAAO inhibitor, 4-F-3-HAA (50 µM). Data are representative of three experiments. (C) An alternative pathway of 3-HAA metabolism can generate CA either by non-enzymatic (oxidation) or enzymatic (such as laccase or ceruloplasmin) processes. (D) Fluorescence [measured in relative fluorescence units (RFUs)] of the AHR-responsive reporter construct in cells incubated with varying concentrations of 3-HAA or CA. Tryptamine was a positive control. *, p

    Article Snippet: For naïve CD4+ T cell sorting, human PBMCs from adult donors or from de-identified cord blood (MD Anderson) were stained with anti-CD3-Alexa700 (BD), anti-CD4-ECD (Invitrogen), anti-CD8-PeCy5.5 (Invitrogen), anti-CD45RA-FITC (BD), anti-CD95-APC (BD), anti-CD25-PE (BD), anti-CCR7-PeCy7 (BD), and anti-CD27-APCCy7 (eBioscience).

    Techniques: Generated, Flow Cytometry, Fluorescence, Construct, Incubation, Positive Control

    3-HKA and 3-HAA promote IL-22 expression in stimulated human CD4 + T cells. (A) Flow cytometric analysis of CD4 + T cells following stimulation of human PBMCs in the presence of 100 µM 3-HKA, 3-HAA, PA, or QA for six days. Data represent at least three independent experiments. (B) Flow cytometric analysis of CD4 + T cells from individual and aggregate donors following stimulation of PBMCs in the presence of increasing concentrations of 3-HAA (µM) for six days. Individual donor data are pooled from at least three independent experiments. Error bars indicate SD. Fold change in IL-22 expression versus vehicle control is statistically different from 1 (Wilcoxon signed rank test; *, p = 0.0312; **, p = 0.0078; N = 8 donors). (C) Flow cytometric analysis of CD4 + T cells following stimulation of human PBMCs in the presence of 3-HAA +/− the AHR antagonist, CH-223191. Data are representative of at least three independent experiments. (D) Comparison of IL-22 production in CD4 + T cells following stimulation of human PBMCs in the presence of DMSO, 3-HKA (50 µM), or 3-HAA (50 µM), with or without an AHR antagonist, N = 6. P values were calculated by Mann-Whitney. Error bars indicate SD.

    Journal: PLoS ONE

    Article Title: Identification of Cinnabarinic Acid as a Novel Endogenous Aryl Hydrocarbon Receptor Ligand That Drives IL-22 Production

    doi: 10.1371/journal.pone.0087877

    Figure Lengend Snippet: 3-HKA and 3-HAA promote IL-22 expression in stimulated human CD4 + T cells. (A) Flow cytometric analysis of CD4 + T cells following stimulation of human PBMCs in the presence of 100 µM 3-HKA, 3-HAA, PA, or QA for six days. Data represent at least three independent experiments. (B) Flow cytometric analysis of CD4 + T cells from individual and aggregate donors following stimulation of PBMCs in the presence of increasing concentrations of 3-HAA (µM) for six days. Individual donor data are pooled from at least three independent experiments. Error bars indicate SD. Fold change in IL-22 expression versus vehicle control is statistically different from 1 (Wilcoxon signed rank test; *, p = 0.0312; **, p = 0.0078; N = 8 donors). (C) Flow cytometric analysis of CD4 + T cells following stimulation of human PBMCs in the presence of 3-HAA +/− the AHR antagonist, CH-223191. Data are representative of at least three independent experiments. (D) Comparison of IL-22 production in CD4 + T cells following stimulation of human PBMCs in the presence of DMSO, 3-HKA (50 µM), or 3-HAA (50 µM), with or without an AHR antagonist, N = 6. P values were calculated by Mann-Whitney. Error bars indicate SD.

    Article Snippet: For naïve CD4+ T cell sorting, human PBMCs from adult donors or from de-identified cord blood (MD Anderson) were stained with anti-CD3-Alexa700 (BD), anti-CD4-ECD (Invitrogen), anti-CD8-PeCy5.5 (Invitrogen), anti-CD45RA-FITC (BD), anti-CD95-APC (BD), anti-CD25-PE (BD), anti-CCR7-PeCy7 (BD), and anti-CD27-APCCy7 (eBioscience).

    Techniques: Expressing, Flow Cytometry, MANN-WHITNEY

    CA increases the differentiation of IL-22 + human CD4 + T cells in vitro . (A) Flow cytometric analysis of CD4 + T cells from human PBMCs stimulated in the presence of DMSO or increasing doses of CA (left). Fold change in IL-22 production in CD4 + T cells from human PBMCs from multiple donors stimulated in the presence of CA versus DMSO control (right panel). Data were analyzed by Wilcoxon signed rank test for significant deviation from a theoretical median of 1.000. *p

    Journal: PLoS ONE

    Article Title: Identification of Cinnabarinic Acid as a Novel Endogenous Aryl Hydrocarbon Receptor Ligand That Drives IL-22 Production

    doi: 10.1371/journal.pone.0087877

    Figure Lengend Snippet: CA increases the differentiation of IL-22 + human CD4 + T cells in vitro . (A) Flow cytometric analysis of CD4 + T cells from human PBMCs stimulated in the presence of DMSO or increasing doses of CA (left). Fold change in IL-22 production in CD4 + T cells from human PBMCs from multiple donors stimulated in the presence of CA versus DMSO control (right panel). Data were analyzed by Wilcoxon signed rank test for significant deviation from a theoretical median of 1.000. *p

    Article Snippet: For naïve CD4+ T cell sorting, human PBMCs from adult donors or from de-identified cord blood (MD Anderson) were stained with anti-CD3-Alexa700 (BD), anti-CD4-ECD (Invitrogen), anti-CD8-PeCy5.5 (Invitrogen), anti-CD45RA-FITC (BD), anti-CD95-APC (BD), anti-CD25-PE (BD), anti-CCR7-PeCy7 (BD), and anti-CD27-APCCy7 (eBioscience).

    Techniques: In Vitro, Flow Cytometry

    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

    Lu AF58786 inhibits phosphorylation of LRRK2, Rab10 and Rab12 in cultured and immune stimulated human PBMCs. ( A ) LRRK2 (pSer910, pSer935, pSer955 and pSer973) phospho-site hits plotted with mean and individual replicate values of the quantified log 10 -transformed CMPD/DMSO ratio as well as the associated q-value of the performed mean rank (n = 5–8). ( B ) Rab10 (pThr73) and Rab12 (pSer106) switch II domain phospho-site hits plotted with mean and individual replicate values of the quantified log 10 -transformed CMPD/DMSO ratio as well as the associated q-value of the performed mean rank (n = 5–8). CMPD, compound (Lu AF58786). DMSO, dimethyl sulfoxide.

    Journal: Scientific Reports

    Article Title: Selective LRRK2 kinase inhibition reduces phosphorylation of endogenous Rab10 and Rab12 in human peripheral mononuclear blood cells

    doi: 10.1038/s41598-017-10501-z

    Figure Lengend Snippet: Lu AF58786 inhibits phosphorylation of LRRK2, Rab10 and Rab12 in cultured and immune stimulated human PBMCs. ( A ) LRRK2 (pSer910, pSer935, pSer955 and pSer973) phospho-site hits plotted with mean and individual replicate values of the quantified log 10 -transformed CMPD/DMSO ratio as well as the associated q-value of the performed mean rank (n = 5–8). ( B ) Rab10 (pThr73) and Rab12 (pSer106) switch II domain phospho-site hits plotted with mean and individual replicate values of the quantified log 10 -transformed CMPD/DMSO ratio as well as the associated q-value of the performed mean rank (n = 5–8). CMPD, compound (Lu AF58786). DMSO, dimethyl sulfoxide.

    Article Snippet: Importantly, this study highlights that phosphorylation of Rab10 in human primary cells can provide an alternative option to pSer910 and pSer935 target engagement assays for determining pharmacodynamic effects of LRRK2 inhibition in human PBMCs.

    Techniques: Cell Culture, Transformation Assay

    Proteomic and phosphoproteomic study to identify LRRK2 kinase activity-dependent substrates in immune-stimulated human PBMCs. ( A ) % viability after 3 DIV of human immune-stimulated PBMCs treated for 24 hrs with or without 100 nM Lu AF58786 (n = 10 donors; 2 conditions). ( B ) PBMC total protein yield (µg) after 3 DIV of human immune-stimulated PBMCs treated for 24 hrs with or without 100 nM Lu AF58786 (n = 10 donors; 2 conditions). ( C ) Experimental set-up with information on mTRAQ labelling and pairing of samples as well as amount of protein used. ( D ) Experimental flow chart of the proteomic studies. Data was analyzed by either one-way ANOVA with Dunnett’s multiple comparisons test or unpaired t-test. Data is presented as means ± SEM. PMA, phorbol 12-myristate 13-acetate; INF-γ, interferon-γ; DMSO, dimethyl sulfoxide.

    Journal: Scientific Reports

    Article Title: Selective LRRK2 kinase inhibition reduces phosphorylation of endogenous Rab10 and Rab12 in human peripheral mononuclear blood cells

    doi: 10.1038/s41598-017-10501-z

    Figure Lengend Snippet: Proteomic and phosphoproteomic study to identify LRRK2 kinase activity-dependent substrates in immune-stimulated human PBMCs. ( A ) % viability after 3 DIV of human immune-stimulated PBMCs treated for 24 hrs with or without 100 nM Lu AF58786 (n = 10 donors; 2 conditions). ( B ) PBMC total protein yield (µg) after 3 DIV of human immune-stimulated PBMCs treated for 24 hrs with or without 100 nM Lu AF58786 (n = 10 donors; 2 conditions). ( C ) Experimental set-up with information on mTRAQ labelling and pairing of samples as well as amount of protein used. ( D ) Experimental flow chart of the proteomic studies. Data was analyzed by either one-way ANOVA with Dunnett’s multiple comparisons test or unpaired t-test. Data is presented as means ± SEM. PMA, phorbol 12-myristate 13-acetate; INF-γ, interferon-γ; DMSO, dimethyl sulfoxide.

    Article Snippet: Importantly, this study highlights that phosphorylation of Rab10 in human primary cells can provide an alternative option to pSer910 and pSer935 target engagement assays for determining pharmacodynamic effects of LRRK2 inhibition in human PBMCs.

    Techniques: Activity Assay, Flow Cytometry

    PFE-360 inhibits LRRK2-Ser935 and Rab10-Thr73 phosphorylation in a concentration-dependent manner in non-stimulated PBMCs from human healthy subjects. Determination of LRRK2 inhibitor IC 50 values based on either Rab10-pThr73 or LRRK2-pSer935 levels in human non-stimulated PBMCs. ( A ) Odyssey CLx scan image showing Western Blot analysis of crude lysates from a pool of PBMCs from two donors treated for 1 hour with concentrations of PFE-360 ranging from 4nM-1µM. in duplicate. LRRK2 and Rab10 immunoreactivity ( red panels ), LRRK2-pSer935 and Rab10-pThr73 immunoreactivity ( green panels ) as well as overlay in non-stimulated human PBMCs. Full-length blots are presented in Supplementary Figure 5 . Non-linear regression plot of percentage ( B ) LRRK2-pSer935 inhibition and ( C ) Rab10-pThr73 as a function of log10-transformed PFE-360 concentration. The experiment was repeated three times and the resulting IC50 determination is summarized in Table 2 .

    Journal: Scientific Reports

    Article Title: Selective LRRK2 kinase inhibition reduces phosphorylation of endogenous Rab10 and Rab12 in human peripheral mononuclear blood cells

    doi: 10.1038/s41598-017-10501-z

    Figure Lengend Snippet: PFE-360 inhibits LRRK2-Ser935 and Rab10-Thr73 phosphorylation in a concentration-dependent manner in non-stimulated PBMCs from human healthy subjects. Determination of LRRK2 inhibitor IC 50 values based on either Rab10-pThr73 or LRRK2-pSer935 levels in human non-stimulated PBMCs. ( A ) Odyssey CLx scan image showing Western Blot analysis of crude lysates from a pool of PBMCs from two donors treated for 1 hour with concentrations of PFE-360 ranging from 4nM-1µM. in duplicate. LRRK2 and Rab10 immunoreactivity ( red panels ), LRRK2-pSer935 and Rab10-pThr73 immunoreactivity ( green panels ) as well as overlay in non-stimulated human PBMCs. Full-length blots are presented in Supplementary Figure 5 . Non-linear regression plot of percentage ( B ) LRRK2-pSer935 inhibition and ( C ) Rab10-pThr73 as a function of log10-transformed PFE-360 concentration. The experiment was repeated three times and the resulting IC50 determination is summarized in Table 2 .

    Article Snippet: Importantly, this study highlights that phosphorylation of Rab10 in human primary cells can provide an alternative option to pSer910 and pSer935 target engagement assays for determining pharmacodynamic effects of LRRK2 inhibition in human PBMCs.

    Techniques: Concentration Assay, Western Blot, Inhibition, Transformation Assay

    LRRK2 kinase-activity dependent LRRK2-Ser935, Rab10-Thr73 and Rab12-Ser106 phosphorylation in cultured and immune stimulated human PBMCs. Quantification of ( A , B ) normalized LRRK2-pSer935/total LRRK2 ( C , D ) Rab10-pThr73/total Rab10 and ( E , F ) Rab12-pSer106/total Rab10 ratios in cultured and immune stimulated human PBMCs treated with either DMSO, 100 nM Lu AF58786 or 100 nM PFE-360 (n = 10 donors; each donor in 3 conditions at left panel , 1hr and right panel , 24hrs). Data was analyzed by one-way ANOVA with Holm-Sidak’s multiple comparisons test. Data is presented as DMSO-normalized means ± SEM; p-values presented are vs. DMSO.

    Journal: Scientific Reports

    Article Title: Selective LRRK2 kinase inhibition reduces phosphorylation of endogenous Rab10 and Rab12 in human peripheral mononuclear blood cells

    doi: 10.1038/s41598-017-10501-z

    Figure Lengend Snippet: LRRK2 kinase-activity dependent LRRK2-Ser935, Rab10-Thr73 and Rab12-Ser106 phosphorylation in cultured and immune stimulated human PBMCs. Quantification of ( A , B ) normalized LRRK2-pSer935/total LRRK2 ( C , D ) Rab10-pThr73/total Rab10 and ( E , F ) Rab12-pSer106/total Rab10 ratios in cultured and immune stimulated human PBMCs treated with either DMSO, 100 nM Lu AF58786 or 100 nM PFE-360 (n = 10 donors; each donor in 3 conditions at left panel , 1hr and right panel , 24hrs). Data was analyzed by one-way ANOVA with Holm-Sidak’s multiple comparisons test. Data is presented as DMSO-normalized means ± SEM; p-values presented are vs. DMSO.

    Article Snippet: Importantly, this study highlights that phosphorylation of Rab10 in human primary cells can provide an alternative option to pSer910 and pSer935 target engagement assays for determining pharmacodynamic effects of LRRK2 inhibition in human PBMCs.

    Techniques: Activity Assay, Cell Culture

    Acute LRRK2 inhibition with PFE-360 reduces LRRK2-pSer935 and Rab10-Thr73 phosphorylation in non-stimulated PBMCs from human healthy subjects. Odyssey CLx scan Western Blot images showing ( A ) LRRK2 and ( C ) Rab10 immunoreactivity (red panels), LRRK2-pSer935 and Rab10-pThr73 immunoreactivity (green panels) as well as overlay in non-stimulated human PBMCs treated with 2 µM PFE-360. Full-length blots are presented in Supplementary Figure 4 . Quantification of ( B ) relative LRRK2-pSer935/total LRRK2 ratio and ( D ) relative Rab10-pThr73/total Rab10 ratio (n = 6 donors; 2 conditions). Data was analyzed by paired t-test. Data is presented as means ± SEM; ****p

    Journal: Scientific Reports

    Article Title: Selective LRRK2 kinase inhibition reduces phosphorylation of endogenous Rab10 and Rab12 in human peripheral mononuclear blood cells

    doi: 10.1038/s41598-017-10501-z

    Figure Lengend Snippet: Acute LRRK2 inhibition with PFE-360 reduces LRRK2-pSer935 and Rab10-Thr73 phosphorylation in non-stimulated PBMCs from human healthy subjects. Odyssey CLx scan Western Blot images showing ( A ) LRRK2 and ( C ) Rab10 immunoreactivity (red panels), LRRK2-pSer935 and Rab10-pThr73 immunoreactivity (green panels) as well as overlay in non-stimulated human PBMCs treated with 2 µM PFE-360. Full-length blots are presented in Supplementary Figure 4 . Quantification of ( B ) relative LRRK2-pSer935/total LRRK2 ratio and ( D ) relative Rab10-pThr73/total Rab10 ratio (n = 6 donors; 2 conditions). Data was analyzed by paired t-test. Data is presented as means ± SEM; ****p

    Article Snippet: Importantly, this study highlights that phosphorylation of Rab10 in human primary cells can provide an alternative option to pSer910 and pSer935 target engagement assays for determining pharmacodynamic effects of LRRK2 inhibition in human PBMCs.

    Techniques: Inhibition, Western Blot

    LRRK2 expression and inhibition in cultured human PBMCs immune stimulated with PMA and interferon- γ . ( A ) Schematic representation of the three different treatment conditions. ( B ) Chemical structure of the LRRK2 inhibitor Lu AF58786. ( C ) Determination of LRRK2, G2019S and A2016T IC 50 values using the Odyssey CLx 96-well ICW assay (mean IC 50 , n = 3 experiments). ( D ) Total PBMC yield obtained from human healthy donors after 3 days in vitro (3 DIV) (n = 10 donors; 3 different conditions). ( E ) % viability of human PBMCs (n = 10 donors; 3 different conditions) after 3 DIV. ( F ) Odyssey CLx scan image of Western Blot used for estimation of LRRK2 protein levels (total LRRK2) and LRRK2 phosphorylation (pSer935). The concentration of Lu AF58786 was 100 nM. Full-length blots are presented in Supplementary Figure 1 . ( G ) Quantification of total LRRK2 levels (raw signal). ( H ) Relative LRRK2-pSer935 phosphorylation (pSer935/total LRRK2). Data was analyzed by one-way ANOVA with Dunnett’s multiple comparisons test. Data presented as means ± SEM; ****p

    Journal: Scientific Reports

    Article Title: Selective LRRK2 kinase inhibition reduces phosphorylation of endogenous Rab10 and Rab12 in human peripheral mononuclear blood cells

    doi: 10.1038/s41598-017-10501-z

    Figure Lengend Snippet: LRRK2 expression and inhibition in cultured human PBMCs immune stimulated with PMA and interferon- γ . ( A ) Schematic representation of the three different treatment conditions. ( B ) Chemical structure of the LRRK2 inhibitor Lu AF58786. ( C ) Determination of LRRK2, G2019S and A2016T IC 50 values using the Odyssey CLx 96-well ICW assay (mean IC 50 , n = 3 experiments). ( D ) Total PBMC yield obtained from human healthy donors after 3 days in vitro (3 DIV) (n = 10 donors; 3 different conditions). ( E ) % viability of human PBMCs (n = 10 donors; 3 different conditions) after 3 DIV. ( F ) Odyssey CLx scan image of Western Blot used for estimation of LRRK2 protein levels (total LRRK2) and LRRK2 phosphorylation (pSer935). The concentration of Lu AF58786 was 100 nM. Full-length blots are presented in Supplementary Figure 1 . ( G ) Quantification of total LRRK2 levels (raw signal). ( H ) Relative LRRK2-pSer935 phosphorylation (pSer935/total LRRK2). Data was analyzed by one-way ANOVA with Dunnett’s multiple comparisons test. Data presented as means ± SEM; ****p

    Article Snippet: Importantly, this study highlights that phosphorylation of Rab10 in human primary cells can provide an alternative option to pSer910 and pSer935 target engagement assays for determining pharmacodynamic effects of LRRK2 inhibition in human PBMCs.

    Techniques: Expressing, Inhibition, Cell Culture, In Vitro, Western Blot, Concentration Assay

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

    Journal: Annals of Clinical and Translational Neurology

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

    doi: 10.1002/acn3.152

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

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

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

    HIV-1 infection in humanized mice passively infused with b12-IgA or b12-IgG isotype. (A) Neutralization activity of anti-HIV human mAb b12-IgA. Reporter cell line TZM-bl cells that express CD4, CXCR4, CCR5, and a Tat-responsive reporter gene for luciferase were infected with 200 TCID 50 of replication-defective pseudovirus containing Env (SF162.LS) in the presence of various concentrations of anti-HIV monoclonal antibodies. Neutralization activity was measured by the reduction in luciferase reporter gene expression after a single round of pseudovirus infection in TZM-bl cells in triplicate. b12-IgG1 and the recombinant b12-IgA2 were compared at indicated concentrations. (B) Antibody level in circulation after passive transfer. NSG-hu mice were injected intravenously with various concentrations of purified b12-IgA2 (pIgA: mIgA = 1:1, mass ratio) or b12-IgG1 and the blood was collected after 4 hours. The plasma antibody concentrations were measured using ELISA. (n = 4-6, mean ± SEM). (C-D) Concentrations of b12 antibodies in plasma (C) or genital secretions (D) at the time of challenge. NSG-hu mice were injected intravenously with either 200 μg of b12-IgA2 or 20 μg of b12-IgG1 per mouse. Blood and genital secretions were collected after 4 hours. (E-I) Peripheral CD4 + T cell loss after HIV-1 challenge in NSG-hu mice injected with different b12 antibody isotypes; (E) purified human IgG/κ control antibody (hIgG/κ); (F) b12-IgA2 includes both monomeric and polymeric IgA as described in (B), (b12IgA2 [M+P]); (G) b12-IgG1; (H) b12-IgA2 monomer only (b12IgA2 [M]). Mice were challenged intravaginally with HIV-1 JR-CSF . (I) Average percent CD4 + T cells in CD3 T cells in PBMCs at each time points (n = 5-9). Statistical analysis was performed using unpaired t test. (* P = .0313 and *** P

    Journal: Blood

    Article Title: Inhibitory effect of HIV-specific neutralizing IgA on mucosal transmission of HIV in humanized mice

    doi: 10.1182/blood-2012-04-422303

    Figure Lengend Snippet: HIV-1 infection in humanized mice passively infused with b12-IgA or b12-IgG isotype. (A) Neutralization activity of anti-HIV human mAb b12-IgA. Reporter cell line TZM-bl cells that express CD4, CXCR4, CCR5, and a Tat-responsive reporter gene for luciferase were infected with 200 TCID 50 of replication-defective pseudovirus containing Env (SF162.LS) in the presence of various concentrations of anti-HIV monoclonal antibodies. Neutralization activity was measured by the reduction in luciferase reporter gene expression after a single round of pseudovirus infection in TZM-bl cells in triplicate. b12-IgG1 and the recombinant b12-IgA2 were compared at indicated concentrations. (B) Antibody level in circulation after passive transfer. NSG-hu mice were injected intravenously with various concentrations of purified b12-IgA2 (pIgA: mIgA = 1:1, mass ratio) or b12-IgG1 and the blood was collected after 4 hours. The plasma antibody concentrations were measured using ELISA. (n = 4-6, mean ± SEM). (C-D) Concentrations of b12 antibodies in plasma (C) or genital secretions (D) at the time of challenge. NSG-hu mice were injected intravenously with either 200 μg of b12-IgA2 or 20 μg of b12-IgG1 per mouse. Blood and genital secretions were collected after 4 hours. (E-I) Peripheral CD4 + T cell loss after HIV-1 challenge in NSG-hu mice injected with different b12 antibody isotypes; (E) purified human IgG/κ control antibody (hIgG/κ); (F) b12-IgA2 includes both monomeric and polymeric IgA as described in (B), (b12IgA2 [M+P]); (G) b12-IgG1; (H) b12-IgA2 monomer only (b12IgA2 [M]). Mice were challenged intravaginally with HIV-1 JR-CSF . (I) Average percent CD4 + T cells in CD3 T cells in PBMCs at each time points (n = 5-9). Statistical analysis was performed using unpaired t test. (* P = .0313 and *** P

    Article Snippet: For passive antibody transfer experiments, NOD.Cg- Prkd scid IL2rg tm1Wjl /SzJ (NOD/SCID IL2r γ−/− , NSG) mice were transplanted intraperitoneally with 3 × 106 human PBMCs (AllCells) that were activated by PHA (5 μg/mL) for 4 days before transplantation (NSG-hu mice).

    Techniques: Infection, Mouse Assay, Neutralization, Activity Assay, Luciferase, Expressing, Recombinant, Injection, Purification, Enzyme-linked Immunosorbent Assay

    The performance of BREM-SC with the public human PBMC CITE-Seq dataset (from 10X Genomics). The UMAP projection of cells are colored by the ground truth ( 4A ) and BREM-SC clustering results ( 4B ).

    Journal: bioRxiv

    Article Title: BREM-SC: A Bayesian Random Effects Mixture Model for Joint Clustering Single Cell Multi-omics Data

    doi: 10.1101/2020.01.18.911461

    Figure Lengend Snippet: The performance of BREM-SC with the public human PBMC CITE-Seq dataset (from 10X Genomics). The UMAP projection of cells are colored by the ground truth ( 4A ) and BREM-SC clustering results ( 4B ).

    Article Snippet: Analysis of a public human PBMC CITE-Seq dataset To evaluate the clustering performance of BREM-SC on real data, we first used a published human PBMC CITE-Seq dataset downloaded from 10X Genomics website.

    Techniques:

    The performance of BREM-SC for in-house human PBMC CITE-Seq dataset. The UMAP projection of cells are colored by the ground truth ( 6A ) and BREM-SC clustering results ( 6B ).

    Journal: bioRxiv

    Article Title: BREM-SC: A Bayesian Random Effects Mixture Model for Joint Clustering Single Cell Multi-omics Data

    doi: 10.1101/2020.01.18.911461

    Figure Lengend Snippet: The performance of BREM-SC for in-house human PBMC CITE-Seq dataset. The UMAP projection of cells are colored by the ground truth ( 6A ) and BREM-SC clustering results ( 6B ).

    Article Snippet: Analysis of a public human PBMC CITE-Seq dataset To evaluate the clustering performance of BREM-SC on real data, we first used a published human PBMC CITE-Seq dataset downloaded from 10X Genomics website.

    Techniques: