Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Vasculogenic Conditioning of Peripheral Blood Mononuclear Cells Promotes Endothelial Progenitor Cell Expansion and Phenotype Transition of Anti‐Inflammatory Macrophage and T Lymphocyte to Cells With Regenerative Potential

doi: 10.1161/JAHA.113.000743

Figure Lengend Snippet: Characteristics of QQMNCs versus PBMNCs. A, The graph shows total cell counts of PBMNCs isolated from 100 mL of PB and the respective QQMNC counts. B, The left graph indicates linear regression analysis between the cell‐count ratio of QQMNCs versus PBMNCs (2×10 6 cells/well) and PBMNC counts isolated from 100 mL of PB. The right indicates linear regression analysis of cell counts between PBMNCs and QQMNCs per 100 mL of PB. C, Representative pictures of pEPC‐CFU and dEPC‐CFU. Scale bar=500 μm. D, The left and middle graphs are EPC‐CFU counts generated from PBMNCs or QQMNCs per dish (2×10 5 cells/dish) and in 100 mL of PB. The right graph shows the percentage of each EPC‐CFU count versus total EPC‐CFU count per dish. Each column in the graph represents a mean±SE. The white and gray areas in the bar graphs indicate the values of pEPC‐CFU and dEPC‐CFU. E, Linear regression graphs on the interrelation of each EPC‐CFU count per dish in QQMNCs to that in PBMNCs. * P

Article Snippet: Early EPC culture of PBMNCs As previously reported, – early EPCs (eEPCs) were acquired after 7 days of culture of isolated PBMNCs using the EGM‐2‐MV SingleQuots kit (Lonza Walkersville, Inc., Walkersville, MD).

Techniques: Isolation, Cell Counting, Generated

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

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

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

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

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

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

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:

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

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

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

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, we also show that both the abovementioned LRRK2 inhibitor compounds have non-overlapping off-target kinases in human PBMCs at 1 µM concentrations.

Techniques: Cell Culture, Transformation Assay

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, we also show that both the abovementioned LRRK2 inhibitor compounds have non-overlapping off-target kinases in human PBMCs at 1 µM concentrations.

Techniques: Activity Assay, Flow Cytometry

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, we also show that both the abovementioned LRRK2 inhibitor compounds have non-overlapping off-target kinases in human PBMCs at 1 µM concentrations.

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

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, we also show that both the abovementioned LRRK2 inhibitor compounds have non-overlapping off-target kinases in human PBMCs at 1 µM concentrations.

Techniques: Activity Assay, Cell Culture

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, we also show that both the abovementioned LRRK2 inhibitor compounds have non-overlapping off-target kinases in human PBMCs at 1 µM concentrations.

Techniques: Inhibition, Western Blot

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, we also show that both the abovementioned LRRK2 inhibitor compounds have non-overlapping off-target kinases in human PBMCs at 1 µM concentrations.

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

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:

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:

Journal: International Journal of Nanomedicine

Article Title: Synthesis of a hemin-containing copolymer as a novel immunostimulator that induces IFN-gamma production

doi: 10.2147/IJN.S166259

Figure Lengend Snippet: The evaluation of the immunostimulatory effects of NIPAM-hemin. Abbreviations: NIPAM, N -isopropylacrylamide; hemin, ferriprotoporphyrin IX chloride; IFN-γ, interferon-γ; IL, interleukin; NK, natural killer; PBMCs, peripheral blood mononuclear cells; ELISA, enzyme-linked immunosorbent assay; TLRs, toll-like receptors; SEAP, secreted embryonic alkaline phosphatase.

Article Snippet: Stimulation of human PBMCs with NIPAM-hemin Commercially available frozen PBMCs were purchased from Cellular Technology Limited (Shaker Heights, OH, USA).

Techniques: Enzyme-linked Immunosorbent Assay

Journal: International Journal of Nanomedicine

Article Title: Synthesis of a hemin-containing copolymer as a novel immunostimulator that induces IFN-gamma production

doi: 10.2147/IJN.S166259

Figure Lengend Snippet: NIPAM-hemin induced the production of IFN-γ and IL-6, and had a lesser effect on the induction of IL-1β, in PBMCs. PBMCs were stimulated with NIPAM-hemin, hemin, NIPAM, or poly-NIPAM (MW=66,400). The levels of ( A ) IFN-γ, ( B ) IL-6, and ( C ) IL-1β were determined after 48 h. The concentration of each stimulant was 500 μg/mL. Hemin was dissolved in DMSO, while the other compounds were dissolved in sterilized deionized water. Notes: Data are expressed as the mean±SD (n=3). * P

Article Snippet: Stimulation of human PBMCs with NIPAM-hemin Commercially available frozen PBMCs were purchased from Cellular Technology Limited (Shaker Heights, OH, USA).

Techniques: Concentration Assay

Journal: Clinical and Experimental Immunology

Article Title: HDAC inhibitors modulate innate immune responses to micro‐organisms relevant to chronic mucocutaneous candidiasis

doi: 10.1111/cei.13192

Figure Lengend Snippet: Histone deacetylase (HDAC) inhibitors modulate proinflammatory immune response to Candida albicans. (a–c) Peripheral blood mononuclear cells (PBMCs) were isolated from buffy coats provided by Sanquin Blood Bank, Nijmegen, the Netherlands. Cells

Article Snippet: Human PBMCs were isolated from healthy volunteers (Sanquin Blood Bank, Nijmegen, the Netherlands) by density‐gradient centrifugation over Ficoll‐Paque (GE Healthcare, Chicago, IL, USA), as described previously [ ].

Techniques: Histone Deacetylase Assay, Isolation