peripheral blood mononuclear cells pbmcs  (Millipore)


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    Millipore peripheral blood mononuclear cells pbmcs
    4a-b Flow <t>cytometric</t> analysis of changes in IL-33 expression in <t>PBMC</t> after relapse. (a) Profile of IL-33 in patients showing highest level of expression of IL-33 at month 0, month 6–9 and month 12–15 (b) cross sectional analysis of IL-33 levels between MS patients after relapse and in OND patients.
    Peripheral Blood Mononuclear Cells Pbmcs, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 614 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Longitudinal changes in the expression of IL-33 and IL-33 regulated genes in relapsing remitting MS"

    Article Title: Longitudinal changes in the expression of IL-33 and IL-33 regulated genes in relapsing remitting MS

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0208755

    4a-b Flow cytometric analysis of changes in IL-33 expression in PBMC after relapse. (a) Profile of IL-33 in patients showing highest level of expression of IL-33 at month 0, month 6–9 and month 12–15 (b) cross sectional analysis of IL-33 levels between MS patients after relapse and in OND patients.
    Figure Legend Snippet: 4a-b Flow cytometric analysis of changes in IL-33 expression in PBMC after relapse. (a) Profile of IL-33 in patients showing highest level of expression of IL-33 at month 0, month 6–9 and month 12–15 (b) cross sectional analysis of IL-33 levels between MS patients after relapse and in OND patients.

    Techniques Used: Flow Cytometry, Expressing, Mass Spectrometry

    2) Product Images from "Effects of ReadiSorb L-GSH in Altering Granulomatous Responses against Mycobacterium tuberculosis Infection"

    Article Title: Effects of ReadiSorb L-GSH in Altering Granulomatous Responses against Mycobacterium tuberculosis Infection

    Journal: Journal of Clinical Medicine

    doi: 10.3390/jcm7030040

    ( A ) Quantification of Erdman within acidified compartments in PBMCs of healthy individuals treated with L-GSH. Green fluorescent protein (GFP)-labeled M. tb was used with lysotracker red DND99, which labeled acidified compartments, as well as DAPI for nucleus staining. When treated with L-GSH, granulomas showed an increase in bacterial number in acidified compartments compared to the bacteria present in non-acidified compartments. Yellow areas correspond to acidified compartments with GFP-labeled bacteria; ( B ) fluorescent staining of granulomas from T2DM individuals infected with M. tb . GFP-labeled M. tb was used with lysotracker red DND99, which labeled acidified compartments, as well as DAPI for nucleus staining. When treated with L-GSH, granulomas showed increased numbers of M. tb in acidified compartments compared to the bacteria present in non-acidified compartments. Yellow areas correspond to acidified compartments with GFP labeled bacteria present. Data represent means ± SE from five T2DM individuals and eight healthy individuals. * p
    Figure Legend Snippet: ( A ) Quantification of Erdman within acidified compartments in PBMCs of healthy individuals treated with L-GSH. Green fluorescent protein (GFP)-labeled M. tb was used with lysotracker red DND99, which labeled acidified compartments, as well as DAPI for nucleus staining. When treated with L-GSH, granulomas showed an increase in bacterial number in acidified compartments compared to the bacteria present in non-acidified compartments. Yellow areas correspond to acidified compartments with GFP-labeled bacteria; ( B ) fluorescent staining of granulomas from T2DM individuals infected with M. tb . GFP-labeled M. tb was used with lysotracker red DND99, which labeled acidified compartments, as well as DAPI for nucleus staining. When treated with L-GSH, granulomas showed increased numbers of M. tb in acidified compartments compared to the bacteria present in non-acidified compartments. Yellow areas correspond to acidified compartments with GFP labeled bacteria present. Data represent means ± SE from five T2DM individuals and eight healthy individuals. * p

    Techniques Used: Labeling, Staining, Infection

    ( A ) Survival of M. tb in untreated and L-GSH granulomas of healthy individuals. All samples were separated through density dependent centrifugation from peripheral blood of volunteers and PBMCs were isolated after washes with PBS. In each category 6 × 10 4 bacteria and 6 × 10 5 immune cells were used for an MOI of 0.1:1. CFU counts of granulomas formed from healthy individuals showed a statistically significant decrease at 15 days when treated with L-GSH; ( B ) Survival of M. tb in granulomas of T2DM individuals. All samples were separated through density-dependent centrifugation from peripheral blood of volunteers and PBMCs were isolated after washes with PBS. In each category 6 × 10 4 bacteria and 6 × 10 5 immune cells were used for an MOI of 0.1:1. CFU counts of granulomas formed from individuals with T2DM showed a statistically significant decrease at 15 days when treated with L-GSH. Data represent means ± SE from eight healthy individuals and six T2DM individuals. *** p
    Figure Legend Snippet: ( A ) Survival of M. tb in untreated and L-GSH granulomas of healthy individuals. All samples were separated through density dependent centrifugation from peripheral blood of volunteers and PBMCs were isolated after washes with PBS. In each category 6 × 10 4 bacteria and 6 × 10 5 immune cells were used for an MOI of 0.1:1. CFU counts of granulomas formed from healthy individuals showed a statistically significant decrease at 15 days when treated with L-GSH; ( B ) Survival of M. tb in granulomas of T2DM individuals. All samples were separated through density-dependent centrifugation from peripheral blood of volunteers and PBMCs were isolated after washes with PBS. In each category 6 × 10 4 bacteria and 6 × 10 5 immune cells were used for an MOI of 0.1:1. CFU counts of granulomas formed from individuals with T2DM showed a statistically significant decrease at 15 days when treated with L-GSH. Data represent means ± SE from eight healthy individuals and six T2DM individuals. *** p

    Techniques Used: Centrifugation, Isolation

    ( A ) Hematoxylin and Eosin staining of granulomas from healthy individuals infected with M. tb . There was an aggregation of cells in both treated and untreated groups of granulomas formed by PBMCs of healthy individuals infected with Erdman strain of M. tb . But there was an apparent increase in the size of granulomas when treated with L-GSH; ( B ) Hematoxylin and Eosin staining of granulomas from T2DM individuals infected with M. tb . Granulomas were formed from PBMCs from T2DM patients. There was a great extent of cell aggregation when treated with L-GSH versus untreated. Microscopy work was done with a light microscope at 1000× magnification under oil immersion.
    Figure Legend Snippet: ( A ) Hematoxylin and Eosin staining of granulomas from healthy individuals infected with M. tb . There was an aggregation of cells in both treated and untreated groups of granulomas formed by PBMCs of healthy individuals infected with Erdman strain of M. tb . But there was an apparent increase in the size of granulomas when treated with L-GSH; ( B ) Hematoxylin and Eosin staining of granulomas from T2DM individuals infected with M. tb . Granulomas were formed from PBMCs from T2DM patients. There was a great extent of cell aggregation when treated with L-GSH versus untreated. Microscopy work was done with a light microscope at 1000× magnification under oil immersion.

    Techniques Used: Staining, Infection, Microscopy, Light Microscopy

    3) Product Images from "Nasal Epithelial Cells Activated with Alternaria and House Dust Mite Induce Not Only Th2 but Also Th1 Immune Responses"

    Article Title: Nasal Epithelial Cells Activated with Alternaria and House Dust Mite Induce Not Only Th2 but Also Th1 Immune Responses

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms21082693

    Effects of nasal epithelial cell conditioned media (NECM) on the production of chemical mediators in peripheral blood mononuclear cells. TNF-α and INF-γ productions were significantly increased by NECM treated with Alternaria , Dermatophagoides pteronyssinus (DP), and Dermatophagoides farina (DF). Negative control (NC), nasal epithelial cell-conditioned media (NECM) treated with 100 ug/mL of Alternaria (Alt100), NECM treated with 100 ug/mL of DP (DP100), NECM treated with 100 ug/mL of DF (DF100), * p
    Figure Legend Snippet: Effects of nasal epithelial cell conditioned media (NECM) on the production of chemical mediators in peripheral blood mononuclear cells. TNF-α and INF-γ productions were significantly increased by NECM treated with Alternaria , Dermatophagoides pteronyssinus (DP), and Dermatophagoides farina (DF). Negative control (NC), nasal epithelial cell-conditioned media (NECM) treated with 100 ug/mL of Alternaria (Alt100), NECM treated with 100 ug/mL of DP (DP100), NECM treated with 100 ug/mL of DF (DF100), * p

    Techniques Used: Negative Control

    4) Product Images from "Identification of a Human SOCS1 Polymorphism That Predicts Rheumatoid Arthritis Severity"

    Article Title: Identification of a Human SOCS1 Polymorphism That Predicts Rheumatoid Arthritis Severity

    Journal: Frontiers in Immunology

    doi: 10.3389/fimmu.2020.01336

    Effects of rs4780355-linked genomic DNA features on SOCS1 expression. (A) Peripheral blood mononuclear cells from 18 patients with RA (five T/C heterozygous, seven C/C homozygous and six T/T homozygous for rs4780355) were PHA-activated for 3 h. SOCS1 levels were determined by qRT-PCR (see Methods) before and after 3 h treatment. Values (2 −ΔΔCt ) are normalized to ACTB and the mean value from heterozygous samples at t = 0; p -values from unpaired t -tests are indicated. (B) Luciferase assays. Two independent constructs for each genotype, which included the SOCS1 polyadenylation site and rs4780355 SNP, were used for Jurkat cell transfection; the Renilla /Firefly luciferase ratio was measured 24 h later. (C) miR-498 or control RNA (MISSION® siRNA Universal Negative Control #2, Sigma) were co-transfected with plasmid constructs. Renilla /Firefly ratios obtained with miR-498 were normalized to control miRNA values. Results from two independent clones and triplicate measurements. (D) Two independent clones, including the polyadenylation site, rs4780355 SNP (C or T) and its associated microsatellite [[TTTTC] 3 or [TTTTC] 5 , respectively], were tested for each haplotype in two experiments with triplicate samples. Statistical significance was determined by the Mann-Whitney U test.
    Figure Legend Snippet: Effects of rs4780355-linked genomic DNA features on SOCS1 expression. (A) Peripheral blood mononuclear cells from 18 patients with RA (five T/C heterozygous, seven C/C homozygous and six T/T homozygous for rs4780355) were PHA-activated for 3 h. SOCS1 levels were determined by qRT-PCR (see Methods) before and after 3 h treatment. Values (2 −ΔΔCt ) are normalized to ACTB and the mean value from heterozygous samples at t = 0; p -values from unpaired t -tests are indicated. (B) Luciferase assays. Two independent constructs for each genotype, which included the SOCS1 polyadenylation site and rs4780355 SNP, were used for Jurkat cell transfection; the Renilla /Firefly luciferase ratio was measured 24 h later. (C) miR-498 or control RNA (MISSION® siRNA Universal Negative Control #2, Sigma) were co-transfected with plasmid constructs. Renilla /Firefly ratios obtained with miR-498 were normalized to control miRNA values. Results from two independent clones and triplicate measurements. (D) Two independent clones, including the polyadenylation site, rs4780355 SNP (C or T) and its associated microsatellite [[TTTTC] 3 or [TTTTC] 5 , respectively], were tested for each haplotype in two experiments with triplicate samples. Statistical significance was determined by the Mann-Whitney U test.

    Techniques Used: Expressing, Quantitative RT-PCR, Luciferase, Construct, Transfection, Negative Control, Plasmid Preparation, Clone Assay, MANN-WHITNEY

    5) Product Images from "DNA Methylation Signature of Post-injury Neointimal Cells During Vascular Remodeling in the Rat Balloon Injury Model"

    Article Title: DNA Methylation Signature of Post-injury Neointimal Cells During Vascular Remodeling in the Rat Balloon Injury Model

    Journal: Molecular biology (Los Angeles, Calif.)

    doi: 10.4172/2168-9547.1000163

    Global CpG methylation analysis Averaged CpG methylation rate and the percentage of highly methylated CpGs are shown for neointimal cells, resident VSMCs, PBMCs and BMMCs. The average methylation shows the ratio of the CpG methylated dinucleotides in each sample to the total number of matched reads. The highly methylated percentage shows the percentage of methylated CpGs with b-values > 90%.
    Figure Legend Snippet: Global CpG methylation analysis Averaged CpG methylation rate and the percentage of highly methylated CpGs are shown for neointimal cells, resident VSMCs, PBMCs and BMMCs. The average methylation shows the ratio of the CpG methylated dinucleotides in each sample to the total number of matched reads. The highly methylated percentage shows the percentage of methylated CpGs with b-values > 90%.

    Techniques Used: CpG Methylation Assay, Methylation

    Histograms of methylation percentage per cytosine The distribution of methylation levels (%) across all CpGs is shown for neointimal cells (A), residential VSMCs (B), PBMCs (C) and BMMCs (D). Methylation levels are bimodal denoting that the majority of bases have either high or low methylation.
    Figure Legend Snippet: Histograms of methylation percentage per cytosine The distribution of methylation levels (%) across all CpGs is shown for neointimal cells (A), residential VSMCs (B), PBMCs (C) and BMMCs (D). Methylation levels are bimodal denoting that the majority of bases have either high or low methylation.

    Techniques Used: Methylation

    Hierarchical clustering of the neointimal cells, residential VSMCs, PBMCs and BMMCs using Pearson’s correlation distance and “Wards method”. Pair-wise Pearson’s correlation scores are shown in the chart.
    Figure Legend Snippet: Hierarchical clustering of the neointimal cells, residential VSMCs, PBMCs and BMMCs using Pearson’s correlation distance and “Wards method”. Pair-wise Pearson’s correlation scores are shown in the chart.

    Techniques Used:

    6) Product Images from "Azaspirane (N-N-diethyl-8,8-dipropyl-2-azaspiro [4.5] decane-2-propanamine) inhibits human multiple myeloma cell growth in the bone marrow milieu in vitro and in vivo"

    Article Title: Azaspirane (N-N-diethyl-8,8-dipropyl-2-azaspiro [4.5] decane-2-propanamine) inhibits human multiple myeloma cell growth in the bone marrow milieu in vitro and in vivo

    Journal: Blood

    doi: 10.1182/blood-2004-09-3794

    Azaspirane induces growth inhibition in MM cell lines and patient MM cells. (A) MM.1S (♦), U266 (▪), and RPMI8226 (▴) MM cells; (B) Dex-sensitive MM.1S (♦) and Dex-resistant MM.1R (▪) MM cells; (C) drug-sensitive RPMI8226 (♦), melphalan-resistant RPMI-LR5 (▪), and doxorubicin-resistant RPMI-Dox40 (▴) cells; (D) OPM1 (▴), INA-6 (▪), and MM.1S (♦) cells; (E) freshly isolated tumor cells from patients with MM (n = 3; ▪, , □), as well as (F) peripheral blood mononuclear cells from healthy volunteers (n = 3; ▪, ▴, ♦) were cultured for 48 hours in the presence of azaspirane (0-5 μM). Cell growth was assessed by MTT assay, and data represent mean (± SD) of quadruplicate cultures.
    Figure Legend Snippet: Azaspirane induces growth inhibition in MM cell lines and patient MM cells. (A) MM.1S (♦), U266 (▪), and RPMI8226 (▴) MM cells; (B) Dex-sensitive MM.1S (♦) and Dex-resistant MM.1R (▪) MM cells; (C) drug-sensitive RPMI8226 (♦), melphalan-resistant RPMI-LR5 (▪), and doxorubicin-resistant RPMI-Dox40 (▴) cells; (D) OPM1 (▴), INA-6 (▪), and MM.1S (♦) cells; (E) freshly isolated tumor cells from patients with MM (n = 3; ▪, , □), as well as (F) peripheral blood mononuclear cells from healthy volunteers (n = 3; ▪, ▴, ♦) were cultured for 48 hours in the presence of azaspirane (0-5 μM). Cell growth was assessed by MTT assay, and data represent mean (± SD) of quadruplicate cultures.

    Techniques Used: Inhibition, Isolation, Cell Culture, MTT Assay

    7) Product Images from "Isolated Rearing at Lactation Increases Gut Microbial Diversity and Post-weaning Performance in Pigs"

    Article Title: Isolated Rearing at Lactation Increases Gut Microbial Diversity and Post-weaning Performance in Pigs

    Journal: Frontiers in Microbiology

    doi: 10.3389/fmicb.2018.02889

    Effect of rearing environment and source of nutrients during lactation on absolute CD3+CD4+ T helper cells (A) , CD3+CD8+ cytotoxic T cells (B) , and CD3+CD4+Foxp3+ regulatory T cells (C) in blood [×10 6 /ml] in 4-, 11-, 20-, 32-, and 55-day-old pigs (LS means). One pig from each litter (10 litter per treatments) was selected for blood sampling. The same pigs were sampled throughout the entire study. The whole blood samples (3 mL) were obtained at day 4 (initial), 11, 20 (weanling), 32, 55, and 103 of ages. PBMCs were isolated using Histopaque ® . Two stars refer to ages having significant differences between treatments when compared to other ages at probability value
    Figure Legend Snippet: Effect of rearing environment and source of nutrients during lactation on absolute CD3+CD4+ T helper cells (A) , CD3+CD8+ cytotoxic T cells (B) , and CD3+CD4+Foxp3+ regulatory T cells (C) in blood [×10 6 /ml] in 4-, 11-, 20-, 32-, and 55-day-old pigs (LS means). One pig from each litter (10 litter per treatments) was selected for blood sampling. The same pigs were sampled throughout the entire study. The whole blood samples (3 mL) were obtained at day 4 (initial), 11, 20 (weanling), 32, 55, and 103 of ages. PBMCs were isolated using Histopaque ® . Two stars refer to ages having significant differences between treatments when compared to other ages at probability value

    Techniques Used: Sampling, Isolation

    8) Product Images from "Epigenetic changes around the pX region and spontaneous HTLV-1 transcription are CTCF-independent"

    Article Title: Epigenetic changes around the pX region and spontaneous HTLV-1 transcription are CTCF-independent

    Journal: Wellcome Open Research

    doi: 10.12688/wellcomeopenres.14741.2

    DNA methylation across the body of the HTLV-1 provirus. ( a ) Upper panel: count of CpG dinucleotides in a window of 350 bp in the HTLV-1 reference genome (L36905). Lower panel: schematic diagram of HTLV-1 provirus indicating the two LTRs and the 9 loci examined by MeDIP. ( b ) DNA methylation on the HTLV-1 provirus in the Tax + and Tax – populations from two HTLV-1-infected T cell clones (Clones TBX4B and 11.65). ( c ) DNA methylation on the HTLV-1 provirus in the CADM1 + Tax + and CADM1 + Tax – populations in PBMCs from two unrelated individuals (Patients TDZ and TED). The asterisk (*) indicates that the PCR failed to amplify.
    Figure Legend Snippet: DNA methylation across the body of the HTLV-1 provirus. ( a ) Upper panel: count of CpG dinucleotides in a window of 350 bp in the HTLV-1 reference genome (L36905). Lower panel: schematic diagram of HTLV-1 provirus indicating the two LTRs and the 9 loci examined by MeDIP. ( b ) DNA methylation on the HTLV-1 provirus in the Tax + and Tax – populations from two HTLV-1-infected T cell clones (Clones TBX4B and 11.65). ( c ) DNA methylation on the HTLV-1 provirus in the CADM1 + Tax + and CADM1 + Tax – populations in PBMCs from two unrelated individuals (Patients TDZ and TED). The asterisk (*) indicates that the PCR failed to amplify.

    Techniques Used: DNA Methylation Assay, Methylated DNA Immunoprecipitation, Infection, Clone Assay, Polymerase Chain Reaction

    DNA methylation in the HTLV-1 LTR of patient-derived PBMCs. ( a ) The HTLV-1 LTR sequence (Accession no. L36905) with CpG dinucleotides highlighted in bold. The three TREs are coloured in red; the TATA box is indicated in the rectangle. ( b ) Schematic diagram of HTLV-1 provirus and the regions amplified with indicated sets of primers for bisulfite-sequencing. Sequencing results for each region are shown in the corresponding panels ( c – f ). The three TREs are indicated by red bars. ( c – f ) Schematic representation of DNA methylation for each clone sequenced. Open circles indicate unmethylated cytosine; closed circles methylated cytosine. The numbers indicate the corresponding CpG sites in panel ( a ).
    Figure Legend Snippet: DNA methylation in the HTLV-1 LTR of patient-derived PBMCs. ( a ) The HTLV-1 LTR sequence (Accession no. L36905) with CpG dinucleotides highlighted in bold. The three TREs are coloured in red; the TATA box is indicated in the rectangle. ( b ) Schematic diagram of HTLV-1 provirus and the regions amplified with indicated sets of primers for bisulfite-sequencing. Sequencing results for each region are shown in the corresponding panels ( c – f ). The three TREs are indicated by red bars. ( c – f ) Schematic representation of DNA methylation for each clone sequenced. Open circles indicate unmethylated cytosine; closed circles methylated cytosine. The numbers indicate the corresponding CpG sites in panel ( a ).

    Techniques Used: DNA Methylation Assay, Derivative Assay, Sequencing, Amplification, Methylation Sequencing, Methylation

    Overview of the cell preparations. ( a ) Preparation of Tax + and Tax – populations from PBMCs obtained from HTLV-1-infected patients. PBMCs were stained for CD4, CADM1 and Tax after overnight culture. Tax + and Tax – fractions were collected from the CADM1 + population. ( b ) Preparation of the Tax + and Tax – populations from HTLV-1-infected T cell clones. HTLV-1-infected T cell clones were stained for intracellular Tax and sorted according to Tax expression.
    Figure Legend Snippet: Overview of the cell preparations. ( a ) Preparation of Tax + and Tax – populations from PBMCs obtained from HTLV-1-infected patients. PBMCs were stained for CD4, CADM1 and Tax after overnight culture. Tax + and Tax – fractions were collected from the CADM1 + population. ( b ) Preparation of the Tax + and Tax – populations from HTLV-1-infected T cell clones. HTLV-1-infected T cell clones were stained for intracellular Tax and sorted according to Tax expression.

    Techniques Used: Infection, Staining, Clone Assay, Expressing

    HTLV-1 transcription in two distinct models. a ) Schematic diagram of HTLV-1 provirus inserted in the host genome. The HTLV-1 provirus has two identical LTRs, one at each end of the provirus. As well as genes encoding the canonical retroviral structural components Gag, Pol and Env, the provirus contains a group of regulatory genes in the pX region on the plus-strand. The plus-strand transcripts, represented by tax , are coloured in red, and the minus-strand transcript HBZ in yellow. ( b ) In PBMCs freshly isolated from HTLV-1 carriers, HTLV-1 reactivates and expresses the plus-strand transcripts within a few hours of culture; but these transcripts remain transcriptionally silent for most of the time in vivo . ( c ) In HTLV-1-infected T cell clones cultured in vitro , the promoter activity for plus-strand transcripts shuttles between the on and off state. The plus-strand transcripts are only produced when the promoter activity is on, yielding only a limited fraction of cells that are positive for the plus-strand transcripts at a given time.
    Figure Legend Snippet: HTLV-1 transcription in two distinct models. a ) Schematic diagram of HTLV-1 provirus inserted in the host genome. The HTLV-1 provirus has two identical LTRs, one at each end of the provirus. As well as genes encoding the canonical retroviral structural components Gag, Pol and Env, the provirus contains a group of regulatory genes in the pX region on the plus-strand. The plus-strand transcripts, represented by tax , are coloured in red, and the minus-strand transcript HBZ in yellow. ( b ) In PBMCs freshly isolated from HTLV-1 carriers, HTLV-1 reactivates and expresses the plus-strand transcripts within a few hours of culture; but these transcripts remain transcriptionally silent for most of the time in vivo . ( c ) In HTLV-1-infected T cell clones cultured in vitro , the promoter activity for plus-strand transcripts shuttles between the on and off state. The plus-strand transcripts are only produced when the promoter activity is on, yielding only a limited fraction of cells that are positive for the plus-strand transcripts at a given time.

    Techniques Used: Isolation, In Vivo, Infection, Clone Assay, Cell Culture, In Vitro, Activity Assay, Produced

    Histone modifications and CTCF-binding in the HTLV-1 provirus. Chromatin immunoprecipitation (ChIP) was used to identify ( a ) histone modifications and CTCF-binding in the Tax + and Tax – populations from an HTLV-1-infected T cell clone (TBX4B); and ( b ) histone modifications in the CADM1 + Tax + and CADM1 + Tax – populations from PBMCs obtained from HTLV-1-infected patients (Patients TW and TCD). The horizontal axis indicates the nucleotide position in the full-length HTLV-1 provirus (J02029), and the vertical axis the read depth (arbitrary units). The reads that aligned within either one of the LTRs are greyed out. The black bars on the horizontal axis indicates the LTRs.
    Figure Legend Snippet: Histone modifications and CTCF-binding in the HTLV-1 provirus. Chromatin immunoprecipitation (ChIP) was used to identify ( a ) histone modifications and CTCF-binding in the Tax + and Tax – populations from an HTLV-1-infected T cell clone (TBX4B); and ( b ) histone modifications in the CADM1 + Tax + and CADM1 + Tax – populations from PBMCs obtained from HTLV-1-infected patients (Patients TW and TCD). The horizontal axis indicates the nucleotide position in the full-length HTLV-1 provirus (J02029), and the vertical axis the read depth (arbitrary units). The reads that aligned within either one of the LTRs are greyed out. The black bars on the horizontal axis indicates the LTRs.

    Techniques Used: Binding Assay, Chromatin Immunoprecipitation, Infection

    CTCF occupancy in the HTLV-1 provirus in patient-derived PBMCs. CTCF occupancy was examined by droplet digital PCR following ChIP for CTCF. The experiment was carried out on PBMCs after overnight incubation in vitro . Replicate 1 is obtained from pooled samples of 4 patients (TCR, TEJ, TED and TW) and Replicate 2 from 3 patients (TED, TCR and TEJ).
    Figure Legend Snippet: CTCF occupancy in the HTLV-1 provirus in patient-derived PBMCs. CTCF occupancy was examined by droplet digital PCR following ChIP for CTCF. The experiment was carried out on PBMCs after overnight incubation in vitro . Replicate 1 is obtained from pooled samples of 4 patients (TCR, TEJ, TED and TW) and Replicate 2 from 3 patients (TED, TCR and TEJ).

    Techniques Used: Derivative Assay, Digital PCR, Chromatin Immunoprecipitation, Incubation, In Vitro

    9) Product Images from "Isolated Rearing at Lactation Increases Gut Microbial Diversity and Post-weaning Performance in Pigs"

    Article Title: Isolated Rearing at Lactation Increases Gut Microbial Diversity and Post-weaning Performance in Pigs

    Journal: Frontiers in Microbiology

    doi: 10.3389/fmicb.2018.02889

    Effect of rearing environment and source of nutrients during lactation on absolute CD3+CD4+ T helper cells (A) , CD3+CD8+ cytotoxic T cells (B) , and CD3+CD4+Foxp3+ regulatory T cells (C) in blood [×10 6 /ml] in 4-, 11-, 20-, 32-, and 55-day-old pigs (LS means). One pig from each litter (10 litter per treatments) was selected for blood sampling. The same pigs were sampled throughout the entire study. The whole blood samples (3 mL) were obtained at day 4 (initial), 11, 20 (weanling), 32, 55, and 103 of ages. PBMCs were isolated using Histopaque ® 1077 (Sigma-Aldrich, St. Louis, MO, United States) density gradient centrifugation, counted by hemocytometer, and expressed as blood concentration based on the volume of whole blood used to isolate the PBMCs, the total number of PBMCs isolated, and the proportions of lymphocyte subsets determined by immunofluorescent staining and flow cytometry. Results on day 103 are presented in Supplementary Table S4 . Two stars refer to ages having significant differences between treatments when compared to other ages at probability value
    Figure Legend Snippet: Effect of rearing environment and source of nutrients during lactation on absolute CD3+CD4+ T helper cells (A) , CD3+CD8+ cytotoxic T cells (B) , and CD3+CD4+Foxp3+ regulatory T cells (C) in blood [×10 6 /ml] in 4-, 11-, 20-, 32-, and 55-day-old pigs (LS means). One pig from each litter (10 litter per treatments) was selected for blood sampling. The same pigs were sampled throughout the entire study. The whole blood samples (3 mL) were obtained at day 4 (initial), 11, 20 (weanling), 32, 55, and 103 of ages. PBMCs were isolated using Histopaque ® 1077 (Sigma-Aldrich, St. Louis, MO, United States) density gradient centrifugation, counted by hemocytometer, and expressed as blood concentration based on the volume of whole blood used to isolate the PBMCs, the total number of PBMCs isolated, and the proportions of lymphocyte subsets determined by immunofluorescent staining and flow cytometry. Results on day 103 are presented in Supplementary Table S4 . Two stars refer to ages having significant differences between treatments when compared to other ages at probability value

    Techniques Used: Sampling, Isolation, Gradient Centrifugation, Concentration Assay, Staining, Flow Cytometry, Cytometry

    10) Product Images from "Peripheral Blood B-Cell Death Compensates for Excessive Proliferation in Lymphoid Tissues and Maintains Homeostasis in Bovine Leukemia Virus-Infected Sheep †"

    Article Title: Peripheral Blood B-Cell Death Compensates for Excessive Proliferation in Lymphoid Tissues and Maintains Homeostasis in Bovine Leukemia Virus-Infected Sheep †

    Journal: Journal of Virology

    doi: 10.1128/JVI.01022-06

    B-cell CFSE kinetics in peripheral blood. (A) CFSE was injected into the jugular vein of a control sheep (117), and blood was recovered at different times (before and 1 min, 2 h, and 6 days after injection) from the other jugular vein. PBMCs were isolated by Percoll gradient centrifugation, and fluorescence was measured by flow cytometry of 10,000 cells ( x axis = CFSE; y axis = number of events). (B) One BLV-infected (2091) and one control (2147) sheep were injected intravenously with a bolus of CFSE, and an aliquot of blood was collected by jugular venipuncture before injection and at 1, 6, or 34 days postinjection. PBMCs were purified, and B cells were labeled with the anti-IgM 1H4 monoclonal antibody and stained with a phycoerythrin conjugate. Finally, 10,000 cells were analyzed by flow cytometry ( x axis = CFSE; y axis = B lymphocytes). The percentages of CFSE + B cells within the total B-lymphocyte population are indicated in the upper right quadrants. (C) Kinetics of the CFSE + B-cell population in the peripheral blood of three BLV-infected (2091, 4535, and 4536; solid lines) and three control (2147, 4533, and 4534; dotted lines) sheep. The arrows indicate key times of CFSE kinetics (day 27 and day 83) at which the percentages of labeled B cells reached baseline levels in BLV-infected sheep and in the controls, respectively.
    Figure Legend Snippet: B-cell CFSE kinetics in peripheral blood. (A) CFSE was injected into the jugular vein of a control sheep (117), and blood was recovered at different times (before and 1 min, 2 h, and 6 days after injection) from the other jugular vein. PBMCs were isolated by Percoll gradient centrifugation, and fluorescence was measured by flow cytometry of 10,000 cells ( x axis = CFSE; y axis = number of events). (B) One BLV-infected (2091) and one control (2147) sheep were injected intravenously with a bolus of CFSE, and an aliquot of blood was collected by jugular venipuncture before injection and at 1, 6, or 34 days postinjection. PBMCs were purified, and B cells were labeled with the anti-IgM 1H4 monoclonal antibody and stained with a phycoerythrin conjugate. Finally, 10,000 cells were analyzed by flow cytometry ( x axis = CFSE; y axis = B lymphocytes). The percentages of CFSE + B cells within the total B-lymphocyte population are indicated in the upper right quadrants. (C) Kinetics of the CFSE + B-cell population in the peripheral blood of three BLV-infected (2091, 4535, and 4536; solid lines) and three control (2147, 4533, and 4534; dotted lines) sheep. The arrows indicate key times of CFSE kinetics (day 27 and day 83) at which the percentages of labeled B cells reached baseline levels in BLV-infected sheep and in the controls, respectively.

    Techniques Used: Injection, Isolation, Gradient Centrifugation, Fluorescence, Flow Cytometry, Cytometry, Infection, Purification, Labeling, Staining

    11) Product Images from "Expression of Latent HIV Induced by the Potent HDAC Inhibitor Suberoylanilide Hydroxamic Acid"

    Article Title: Expression of Latent HIV Induced by the Potent HDAC Inhibitor Suberoylanilide Hydroxamic Acid

    Journal: AIDS Research and Human Retroviruses

    doi: 10.1089/aid.2008.0191

    Toxicity of SAHA in seronegative donor PBMC and J89 cells. PBMC or J89 cells were cultured in the absence or presence of SAHA or VPA at the indicated concentrations. MTT assays were performed as described in methods. The percentage of proliferating cells
    Figure Legend Snippet: Toxicity of SAHA in seronegative donor PBMC and J89 cells. PBMC or J89 cells were cultured in the absence or presence of SAHA or VPA at the indicated concentrations. MTT assays were performed as described in methods. The percentage of proliferating cells

    Techniques Used: Cell Culture, MTT Assay

    12) Product Images from "Effects of ReadiSorb L-GSH in Altering Granulomatous Responses against Mycobacterium tuberculosis Infection"

    Article Title: Effects of ReadiSorb L-GSH in Altering Granulomatous Responses against Mycobacterium tuberculosis Infection

    Journal: Journal of Clinical Medicine

    doi: 10.3390/jcm7030040

    ( A ) Quantification of Erdman within acidified compartments in PBMCs of healthy individuals treated with L-GSH. Green fluorescent protein (GFP)-labeled M. tb was used with lysotracker red DND99, which labeled acidified compartments, as well as DAPI for nucleus staining. When treated with L-GSH, granulomas showed an increase in bacterial number in acidified compartments compared to the bacteria present in non-acidified compartments. Yellow areas correspond to acidified compartments with GFP-labeled bacteria; ( B ) fluorescent staining of granulomas from T2DM individuals infected with M. tb . GFP-labeled M. tb was used with lysotracker red DND99, which labeled acidified compartments, as well as DAPI for nucleus staining. When treated with L-GSH, granulomas showed increased numbers of M. tb in acidified compartments compared to the bacteria present in non-acidified compartments. Yellow areas correspond to acidified compartments with GFP labeled bacteria present. Data represent means ± SE from five T2DM individuals and eight healthy individuals. * p
    Figure Legend Snippet: ( A ) Quantification of Erdman within acidified compartments in PBMCs of healthy individuals treated with L-GSH. Green fluorescent protein (GFP)-labeled M. tb was used with lysotracker red DND99, which labeled acidified compartments, as well as DAPI for nucleus staining. When treated with L-GSH, granulomas showed an increase in bacterial number in acidified compartments compared to the bacteria present in non-acidified compartments. Yellow areas correspond to acidified compartments with GFP-labeled bacteria; ( B ) fluorescent staining of granulomas from T2DM individuals infected with M. tb . GFP-labeled M. tb was used with lysotracker red DND99, which labeled acidified compartments, as well as DAPI for nucleus staining. When treated with L-GSH, granulomas showed increased numbers of M. tb in acidified compartments compared to the bacteria present in non-acidified compartments. Yellow areas correspond to acidified compartments with GFP labeled bacteria present. Data represent means ± SE from five T2DM individuals and eight healthy individuals. * p

    Techniques Used: Labeling, Staining, Infection

    ( A ) Survival of M. tb in untreated and L-GSH granulomas of healthy individuals. All samples were separated through density dependent centrifugation from peripheral blood of volunteers and PBMCs were isolated after washes with PBS. In each category 6 × 10 4 bacteria and 6 × 10 5 immune cells were used for an MOI of 0.1:1. CFU counts of granulomas formed from healthy individuals showed a statistically significant decrease at 15 days when treated with L-GSH; ( B ) Survival of M. tb in granulomas of T2DM individuals. All samples were separated through density-dependent centrifugation from peripheral blood of volunteers and PBMCs were isolated after washes with PBS. In each category 6 × 10 4 bacteria and 6 × 10 5 immune cells were used for an MOI of 0.1:1. CFU counts of granulomas formed from individuals with T2DM showed a statistically significant decrease at 15 days when treated with L-GSH. Data represent means ± SE from eight healthy individuals and six T2DM individuals. *** p
    Figure Legend Snippet: ( A ) Survival of M. tb in untreated and L-GSH granulomas of healthy individuals. All samples were separated through density dependent centrifugation from peripheral blood of volunteers and PBMCs were isolated after washes with PBS. In each category 6 × 10 4 bacteria and 6 × 10 5 immune cells were used for an MOI of 0.1:1. CFU counts of granulomas formed from healthy individuals showed a statistically significant decrease at 15 days when treated with L-GSH; ( B ) Survival of M. tb in granulomas of T2DM individuals. All samples were separated through density-dependent centrifugation from peripheral blood of volunteers and PBMCs were isolated after washes with PBS. In each category 6 × 10 4 bacteria and 6 × 10 5 immune cells were used for an MOI of 0.1:1. CFU counts of granulomas formed from individuals with T2DM showed a statistically significant decrease at 15 days when treated with L-GSH. Data represent means ± SE from eight healthy individuals and six T2DM individuals. *** p

    Techniques Used: Centrifugation, Isolation

    ( A ) Hematoxylin and Eosin staining of granulomas from healthy individuals infected with M. tb . There was an aggregation of cells in both treated and untreated groups of granulomas formed by PBMCs of healthy individuals infected with Erdman strain of M. tb . But there was an apparent increase in the size of granulomas when treated with L-GSH; ( B ) Hematoxylin and Eosin staining of granulomas from T2DM individuals infected with M. tb . Granulomas were formed from PBMCs from T2DM patients. There was a great extent of cell aggregation when treated with L-GSH versus untreated. Microscopy work was done with a light microscope at 1000× magnification under oil immersion.
    Figure Legend Snippet: ( A ) Hematoxylin and Eosin staining of granulomas from healthy individuals infected with M. tb . There was an aggregation of cells in both treated and untreated groups of granulomas formed by PBMCs of healthy individuals infected with Erdman strain of M. tb . But there was an apparent increase in the size of granulomas when treated with L-GSH; ( B ) Hematoxylin and Eosin staining of granulomas from T2DM individuals infected with M. tb . Granulomas were formed from PBMCs from T2DM patients. There was a great extent of cell aggregation when treated with L-GSH versus untreated. Microscopy work was done with a light microscope at 1000× magnification under oil immersion.

    Techniques Used: Staining, Infection, Microscopy, Light Microscopy

    13) Product Images from "CD8+ clonality is associated with prolonged acute plasma viremia and altered mRNA cytokine profiles during the course of Feline Immunodeficiency Virus infection"

    Article Title: CD8+ clonality is associated with prolonged acute plasma viremia and altered mRNA cytokine profiles during the course of Feline Immunodeficiency Virus infection

    Journal: Veterinary immunology and immunopathology

    doi: 10.1016/j.vetimm.2012.12.005

    Proliferative response of CD8 + T cells to mitogen stimulation PBMCs from FIV-infected PARR + (N = 4) or PARR − (N = 6) cats were CFSE stained, incubated for 72 hours with ConA (5 ug/mL) and IL-2 (100 U/mL) stimulation, and analyzed for proliferative response by flow cytometry. ModFit LT software was used to calculate the proliferation index (PrI) and the stimulation index (SI). Statistics were performed using the Mann-Whitney test with p
    Figure Legend Snippet: Proliferative response of CD8 + T cells to mitogen stimulation PBMCs from FIV-infected PARR + (N = 4) or PARR − (N = 6) cats were CFSE stained, incubated for 72 hours with ConA (5 ug/mL) and IL-2 (100 U/mL) stimulation, and analyzed for proliferative response by flow cytometry. ModFit LT software was used to calculate the proliferation index (PrI) and the stimulation index (SI). Statistics were performed using the Mann-Whitney test with p

    Techniques Used: Infection, Staining, Incubation, Flow Cytometry, Cytometry, Software, MANN-WHITNEY

    14) Product Images from "Partially Randomized, Non-Blinded Trial of DNA and MVA Therapeutic Vaccines Based on Hepatitis B Virus Surface Protein for Chronic HBV Infection"

    Article Title: Partially Randomized, Non-Blinded Trial of DNA and MVA Therapeutic Vaccines Based on Hepatitis B Virus Surface Protein for Chronic HBV Infection

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0014626

    Background and net ELISpot results normalized to per million PBMCs. where RN10 is the average of the spots from the two negative control wells. The summation is over all 14 wells in the plate layout which contained overlapping pooled HBsAg peptides for each volunteer, and the factor of 1/2 normalizes for each peptide appearing twice in the matrix layout. Because of the 14 summations the effect of a slightly low or slightly high background (measured over only 2 wells) gets amplified in the final net spots count. The immune response would be expected to be strongest in the post-treatment time interval and to wane in the follow-up period, but in fact variability was often high in the follow-up period. This may reflect non-specific immune activation due to other maladies such as malaria or to the natural course of engaging a chronic HBV infection.
    Figure Legend Snippet: Background and net ELISpot results normalized to per million PBMCs. where RN10 is the average of the spots from the two negative control wells. The summation is over all 14 wells in the plate layout which contained overlapping pooled HBsAg peptides for each volunteer, and the factor of 1/2 normalizes for each peptide appearing twice in the matrix layout. Because of the 14 summations the effect of a slightly low or slightly high background (measured over only 2 wells) gets amplified in the final net spots count. The immune response would be expected to be strongest in the post-treatment time interval and to wane in the follow-up period, but in fact variability was often high in the follow-up period. This may reflect non-specific immune activation due to other maladies such as malaria or to the natural course of engaging a chronic HBV infection.

    Techniques Used: Enzyme-linked Immunospot, Negative Control, Amplification, Activation Assay, Infection

    15) Product Images from "Progranulin and a Five Transmembrane Domain-Containing Receptor-like Gene Are the Key Components in Receptor Activator of Nuclear Factor κB (RANK)-dependent Formation of Multinucleated Osteoclasts *"

    Article Title: Progranulin and a Five Transmembrane Domain-Containing Receptor-like Gene Are the Key Components in Receptor Activator of Nuclear Factor κB (RANK)-dependent Formation of Multinucleated Osteoclasts *

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M114.608786

    Effects of PGRN on the differentiation of human OCs and PGRN as a predictor of osteoporosis. A , HBMCs and PBMCs were differentiated with RANKL in the absence or presence of PGRN. TRAP + MNCs were counted as described above. B , four sh-RNAs targeting human
    Figure Legend Snippet: Effects of PGRN on the differentiation of human OCs and PGRN as a predictor of osteoporosis. A , HBMCs and PBMCs were differentiated with RANKL in the absence or presence of PGRN. TRAP + MNCs were counted as described above. B , four sh-RNAs targeting human

    Techniques Used:

    16) Product Images from "Celiac Disease Monocytes Induce a Barrier Defect in Intestinal Epithelial Cells"

    Article Title: Celiac Disease Monocytes Induce a Barrier Defect in Intestinal Epithelial Cells

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms20225597

    Epithelial barrier function after co-culture of intestinal epithelial cells (IECs) with monocytes derived from celiac disease patients. After peripheral blood mononuclear cell (PBMC) isolation and CD14+ cell-sorting, epithelial cells were co-cultured with monocytes from healthy donors. celiac patients on a gluten-free diet (GFD), or AC (active celiac disease (CeD)) patients. Subsequently, the TER was measured after 48 h of co-culture (% of TER prior to addition of monocytes). Mean of n = 36 (healthy donors), n = 15 (GFD), and n = 20 (AC) individual filters measurements. Monocytes used for these experiments were isolated from n = 8 (healthy donors), n = 4 (GFD) and n = 5 (active CeD). Mann-Whitney U * p
    Figure Legend Snippet: Epithelial barrier function after co-culture of intestinal epithelial cells (IECs) with monocytes derived from celiac disease patients. After peripheral blood mononuclear cell (PBMC) isolation and CD14+ cell-sorting, epithelial cells were co-cultured with monocytes from healthy donors. celiac patients on a gluten-free diet (GFD), or AC (active celiac disease (CeD)) patients. Subsequently, the TER was measured after 48 h of co-culture (% of TER prior to addition of monocytes). Mean of n = 36 (healthy donors), n = 15 (GFD), and n = 20 (AC) individual filters measurements. Monocytes used for these experiments were isolated from n = 8 (healthy donors), n = 4 (GFD) and n = 5 (active CeD). Mann-Whitney U * p

    Techniques Used: Co-Culture Assay, Derivative Assay, Isolation, FACS, Cell Culture, MANN-WHITNEY

    Increased levels of pro-inflammatory cytokines in the supernatant of monocytes derived from celiac disease patients. ( A ) PBMCs were isolated from 10 controls, 10 GFD, and 4 active CeD patients, and monocytes were sorted using CD14 MACS MicroBeads. Subsequently, cells were kept in the incubator for 24 h in the presence of GM-CSF (10 ng/mL). Supernatants of monocyte cultures were collected and cytokine levels were determined. Data are illustrated as a heat map revealing color-coded concentrations of cytokines (green: low concentration; red: high concentration). ( B – F ) Results for individual cytokine measurements are shown: Interleukin- (IL-)8, IL-10, IL-1β, TNF-α, IL-6, and MCP-1. Mean values ± SEM are shown. Mann–Whitney test: *, p
    Figure Legend Snippet: Increased levels of pro-inflammatory cytokines in the supernatant of monocytes derived from celiac disease patients. ( A ) PBMCs were isolated from 10 controls, 10 GFD, and 4 active CeD patients, and monocytes were sorted using CD14 MACS MicroBeads. Subsequently, cells were kept in the incubator for 24 h in the presence of GM-CSF (10 ng/mL). Supernatants of monocyte cultures were collected and cytokine levels were determined. Data are illustrated as a heat map revealing color-coded concentrations of cytokines (green: low concentration; red: high concentration). ( B – F ) Results for individual cytokine measurements are shown: Interleukin- (IL-)8, IL-10, IL-1β, TNF-α, IL-6, and MCP-1. Mean values ± SEM are shown. Mann–Whitney test: *, p

    Techniques Used: Derivative Assay, Isolation, Magnetic Cell Separation, Concentration Assay, MANN-WHITNEY

    Expression of surface markers in peripheral monocytes from celiac patients. PBMCs were isolated from 10 controls, 10 GFD and 4 active CeD (AC) patients and sorted for CD14. Subsequently, cells were cultured in the presence of granulocyte macrophage colony stimulating factor (GM-CSF; 10 ng/ml) for 24 h and evaluated by flow cytometry. ( A ) Gating strategy used to determine surface marker expression. The stepwise gating approach is highlighted by various steps of analysis that are interconnected by red arrows. Representative plots from a healthy control are shown. ( B – G ) Results for the expression of single surface markers are shown. Each dot represents the surface marker expression result of a single patient. Mean values ± standard error of the mean (SEM) are shown. The Mann–Whitney U test revealed no significant differences.
    Figure Legend Snippet: Expression of surface markers in peripheral monocytes from celiac patients. PBMCs were isolated from 10 controls, 10 GFD and 4 active CeD (AC) patients and sorted for CD14. Subsequently, cells were cultured in the presence of granulocyte macrophage colony stimulating factor (GM-CSF; 10 ng/ml) for 24 h and evaluated by flow cytometry. ( A ) Gating strategy used to determine surface marker expression. The stepwise gating approach is highlighted by various steps of analysis that are interconnected by red arrows. Representative plots from a healthy control are shown. ( B – G ) Results for the expression of single surface markers are shown. Each dot represents the surface marker expression result of a single patient. Mean values ± standard error of the mean (SEM) are shown. The Mann–Whitney U test revealed no significant differences.

    Techniques Used: Expressing, Isolation, Cell Culture, Flow Cytometry, Cytometry, Marker, MANN-WHITNEY

    17) Product Images from "Trastuzumab Increases HER2 Uptake and Cross-Presentation by Dendritic Cells"

    Article Title: Trastuzumab Increases HER2 Uptake and Cross-Presentation by Dendritic Cells

    Journal: Cancer research

    doi: 10.1158/0008-5472.CAN-16-2774

    Trastuzumab increases cross-presentation of E75 by DCs. A, mature DCs were cultured with SKBR3 cells +/− trastzumab (10 µg/mL) for 24 hours after which the cell surface was stained for DC markers (CD11c + , HLA-DR + ) as well as an Fab targeting E75 peptide/HLA-A2. Cells were analyzed with flow cytometry and results are expressed as average E75/HLA-A2 Fab MFI. DCs cultured with SKBR3 cells + trastuzmab resulted in increased E75 cross-presentation compared to DCs cultured with SKBR3 cells alone. B, C mature DCs were cultured with SKBR3 cells +/− trastuzumab (10–40 µg/mL) to activate and expand E75-CTLs from HLA-A2 + PBMCs. After 1 week of co-culture, enumeration of E75-CTLs was performed using an E75/HLA-A2 dextramer. Results are expressed as percent CD8 + /E75 + cells from a parent gating of Live/CD16 − /CD19 − /CD56 − /CD14 − /CD3 + /CD8 + cells. DCs that had been cultured with SKBR3 cells + trastuzumab resulted in more efficient expansion of E75-CTLs from donor PBMCs. B, representative scatter plots. C, aggregate results of three experiments. * indicates P
    Figure Legend Snippet: Trastuzumab increases cross-presentation of E75 by DCs. A, mature DCs were cultured with SKBR3 cells +/− trastzumab (10 µg/mL) for 24 hours after which the cell surface was stained for DC markers (CD11c + , HLA-DR + ) as well as an Fab targeting E75 peptide/HLA-A2. Cells were analyzed with flow cytometry and results are expressed as average E75/HLA-A2 Fab MFI. DCs cultured with SKBR3 cells + trastuzmab resulted in increased E75 cross-presentation compared to DCs cultured with SKBR3 cells alone. B, C mature DCs were cultured with SKBR3 cells +/− trastuzumab (10–40 µg/mL) to activate and expand E75-CTLs from HLA-A2 + PBMCs. After 1 week of co-culture, enumeration of E75-CTLs was performed using an E75/HLA-A2 dextramer. Results are expressed as percent CD8 + /E75 + cells from a parent gating of Live/CD16 − /CD19 − /CD56 − /CD14 − /CD3 + /CD8 + cells. DCs that had been cultured with SKBR3 cells + trastuzumab resulted in more efficient expansion of E75-CTLs from donor PBMCs. B, representative scatter plots. C, aggregate results of three experiments. * indicates P

    Techniques Used: Cell Culture, Staining, Flow Cytometry, Cytometry, Co-Culture Assay

    18) Product Images from "First-in-human PeriCord cardiac bioimplant: Scalability and GMP manufacturing of an allogeneic engineered tissue graft"

    Article Title: First-in-human PeriCord cardiac bioimplant: Scalability and GMP manufacturing of an allogeneic engineered tissue graft

    Journal: EBioMedicine

    doi: 10.1016/j.ebiom.2020.102729

    Clinical-grade WJ-MSC characterisation. (a) Representative microscopic image of expanded WJ-MSCs culture. Scale bar = 100 µm. (b) Normal karyotype of a DP, corresponding to a representative cell line. (c) Immunophenotypic characteristics of four clinical-grade cell batches and average expression (mean ± SEM). (d) Immunomodulation potential measured in four batches of clinical-grade WJ-MSCs, as the capacity to inhibit proliferation of polyclonal stimulated lymphocytes. (e) Microscopic images showing stimulated PBMC clumping, and clump reduction when co-cultured with WJ-MSCs. Arrowheads indicate clumps. Scale bars = 100 µm. PBMCs, peripheral blood mononuclear cells; WJ-MSCs, Wharton's jelly mesenchymal stromal cells.
    Figure Legend Snippet: Clinical-grade WJ-MSC characterisation. (a) Representative microscopic image of expanded WJ-MSCs culture. Scale bar = 100 µm. (b) Normal karyotype of a DP, corresponding to a representative cell line. (c) Immunophenotypic characteristics of four clinical-grade cell batches and average expression (mean ± SEM). (d) Immunomodulation potential measured in four batches of clinical-grade WJ-MSCs, as the capacity to inhibit proliferation of polyclonal stimulated lymphocytes. (e) Microscopic images showing stimulated PBMC clumping, and clump reduction when co-cultured with WJ-MSCs. Arrowheads indicate clumps. Scale bars = 100 µm. PBMCs, peripheral blood mononuclear cells; WJ-MSCs, Wharton's jelly mesenchymal stromal cells.

    Techniques Used: Expressing, Cell Culture

    19) Product Images from "Effect of Recombinant ?1-Antitrypsin Fc-Fused (AAT-Fc)Protein on the Inhibition of Inflammatory Cytokine Production and Streptozotocin-Induced Diabetes"

    Article Title: Effect of Recombinant ?1-Antitrypsin Fc-Fused (AAT-Fc)Protein on the Inhibition of Inflammatory Cytokine Production and Streptozotocin-Induced Diabetes

    Journal: Molecular Medicine

    doi: 10.2119/molmed.2012.00308

    Dose-dependent suppression of TNFα-induced IL-6 production in human PBMCs. (A) AAT-Fc inhibited TNFα-induced IL-6 production in a dose-dependent manner, and AAT-Fc used at a concentration of 13 nmol/L abolished IL-6 production completely.
    Figure Legend Snippet: Dose-dependent suppression of TNFα-induced IL-6 production in human PBMCs. (A) AAT-Fc inhibited TNFα-induced IL-6 production in a dose-dependent manner, and AAT-Fc used at a concentration of 13 nmol/L abolished IL-6 production completely.

    Techniques Used: Concentration Assay

    20) Product Images from "The HIV-1 Nef Protein Interacts with two components of the 40S small ribosomal subunit, the RPS10 protein and the 18S rRNA"

    Article Title: The HIV-1 Nef Protein Interacts with two components of the 40S small ribosomal subunit, the RPS10 protein and the 18S rRNA

    Journal: Virology Journal

    doi: 10.1186/1743-422X-9-103

    Expression of RPS10/rNef complexes in primary PBMCs, PBLs and MDMs treated with rNef. ( A ) TotalTotal cellular extracts from PBMCs treated with rNef (100 ng/ml) for 2 hours were immunoprecipitated with an anti-RPS10 mAb or an anti-Nef mAb. Immunoprecipitated material was analyzed by western blotting with an anti-Nef mAb or an anti-RPS10 mAb. Results are representative of three independent experiments. ( B ) Cytoplasmic and nuclear extracts from several cell lines (Vero cells, U937 cells), PBLs and MDMs treated with rNef (100 ng/ml) for 2 hours were immunoprecipitated with an anti-RPS10 mAb. Immunoprecipitated material was analyzed by western blotting with an anti-Nef mAb. The controls are input controls from total cell lysates (for IP RPS10 and mouse IgG control). Results are representative of three independent experiments. ( C ) Cytoplasmic and nuclear extracts from U937 cells treated with increasing concentrations of rNef (100–1500 ng/ml) for 2 hours or mock-treated were immunoprecipitated with an anti-RPS10 mAb. Immunoprecipitated material was analyzed by western blotting with an anti-Nef mAb. Results are representative of three independent experiments. β-actin and TBP (TATA binding protein) detection represents input loading controls of the lysates which were used in binding reactions.
    Figure Legend Snippet: Expression of RPS10/rNef complexes in primary PBMCs, PBLs and MDMs treated with rNef. ( A ) TotalTotal cellular extracts from PBMCs treated with rNef (100 ng/ml) for 2 hours were immunoprecipitated with an anti-RPS10 mAb or an anti-Nef mAb. Immunoprecipitated material was analyzed by western blotting with an anti-Nef mAb or an anti-RPS10 mAb. Results are representative of three independent experiments. ( B ) Cytoplasmic and nuclear extracts from several cell lines (Vero cells, U937 cells), PBLs and MDMs treated with rNef (100 ng/ml) for 2 hours were immunoprecipitated with an anti-RPS10 mAb. Immunoprecipitated material was analyzed by western blotting with an anti-Nef mAb. The controls are input controls from total cell lysates (for IP RPS10 and mouse IgG control). Results are representative of three independent experiments. ( C ) Cytoplasmic and nuclear extracts from U937 cells treated with increasing concentrations of rNef (100–1500 ng/ml) for 2 hours or mock-treated were immunoprecipitated with an anti-RPS10 mAb. Immunoprecipitated material was analyzed by western blotting with an anti-Nef mAb. Results are representative of three independent experiments. β-actin and TBP (TATA binding protein) detection represents input loading controls of the lysates which were used in binding reactions.

    Techniques Used: Expressing, Immunoprecipitation, Western Blot, Binding Assay

    21) Product Images from "Combining flagellin and human β-defensin-3 to combat bacterial infections"

    Article Title: Combining flagellin and human β-defensin-3 to combat bacterial infections

    Journal: Frontiers in Microbiology

    doi: 10.3389/fmicb.2014.00673

    Effect of F protein on expression levels of IL12A and IFNγ in CD4+ T cells from healthy donors. (A–C) Effect of F protein on expression levels of IL12A and IFNγ in naïve and activated CD4+ T cells. (A) Experimental design for naïve cells – CD4+ T cells were isolated from peripheral blood of healthy donors using CD4+ magnetic-beads. CD4+ T cells were seeded and incubated with the F protein (2 μg/ml) for 18 or 24 h. (B) Experimental design for activated cells – mononuclear cells were activated by PHA (20 μg/ml) for 72 h and then grown with IL2 (10 units/ml recombinant IL-2). Following activation, CD4+ T cells were separated using CD4 magnetic beads. CD4+ T cells were seeded and incubated with the F protein (2 μg/ml) for 24 or 48 h. Following the incubation, cDNA samples were prepared and subjected to real-time-PCR analysis using SYBR green reagent (as described in Methods). (C) Changes in IFNγ-mRNA and IL12A-mRNA levels upon F stimuli, compared to control cells. I = one F stimulation. Data represents the mean ± SD of 3–5 separate experiments. (D,E) Effect of the F protein on expression levels of IL12A and IFNγ in naïve CD4+ T cells after triggering all mononuclear cells. (D) Experimental design – Naïve mononuclear cells were incubated with flagellin for different time periods, after which CD4+ T cells were separated using CD4 magnetic beads. The separated CD4+ T cells were taken and cDNA samples were prepared and subjected to real time-PCR analysis using SYBR green reagent (as described in Methods). (E) The changes of IFNγ-mRNA and IL12A-mRNA in CD4+ T cells levels upon flagellin stimuli in the presence of mononuclear cells after 2, 6, 12, 24, and 36 h. I = one F stimulation. Data represents the mean ± SD of 3–5 separate experiments. (F,G) Effect of the F protein on expression levels of IL12A and IFNγ in naïve CD4+ T cells after triggering the mononuclear cells three times. (F) Experimental design – Naïve mononuclear cells were triggered three times with flagellin for 2 or 12 h stimuli. Following stimulation, CD4+ T cells were separated using cd4 magnetic beads. The separated CD4+ T cells were removed and cDNA samples were prepared and subjected to real time PCR analysis using SYBR green reagent (as described in Methods). (G) Changes of IFNγ- and IL12A mRNA levels upon repeated flagellin stimulations for 2 or 12 h each. III = three F stimulations. Data represents the mean ± SD of 3–5 separate experiments.
    Figure Legend Snippet: Effect of F protein on expression levels of IL12A and IFNγ in CD4+ T cells from healthy donors. (A–C) Effect of F protein on expression levels of IL12A and IFNγ in naïve and activated CD4+ T cells. (A) Experimental design for naïve cells – CD4+ T cells were isolated from peripheral blood of healthy donors using CD4+ magnetic-beads. CD4+ T cells were seeded and incubated with the F protein (2 μg/ml) for 18 or 24 h. (B) Experimental design for activated cells – mononuclear cells were activated by PHA (20 μg/ml) for 72 h and then grown with IL2 (10 units/ml recombinant IL-2). Following activation, CD4+ T cells were separated using CD4 magnetic beads. CD4+ T cells were seeded and incubated with the F protein (2 μg/ml) for 24 or 48 h. Following the incubation, cDNA samples were prepared and subjected to real-time-PCR analysis using SYBR green reagent (as described in Methods). (C) Changes in IFNγ-mRNA and IL12A-mRNA levels upon F stimuli, compared to control cells. I = one F stimulation. Data represents the mean ± SD of 3–5 separate experiments. (D,E) Effect of the F protein on expression levels of IL12A and IFNγ in naïve CD4+ T cells after triggering all mononuclear cells. (D) Experimental design – Naïve mononuclear cells were incubated with flagellin for different time periods, after which CD4+ T cells were separated using CD4 magnetic beads. The separated CD4+ T cells were taken and cDNA samples were prepared and subjected to real time-PCR analysis using SYBR green reagent (as described in Methods). (E) The changes of IFNγ-mRNA and IL12A-mRNA in CD4+ T cells levels upon flagellin stimuli in the presence of mononuclear cells after 2, 6, 12, 24, and 36 h. I = one F stimulation. Data represents the mean ± SD of 3–5 separate experiments. (F,G) Effect of the F protein on expression levels of IL12A and IFNγ in naïve CD4+ T cells after triggering the mononuclear cells three times. (F) Experimental design – Naïve mononuclear cells were triggered three times with flagellin for 2 or 12 h stimuli. Following stimulation, CD4+ T cells were separated using cd4 magnetic beads. The separated CD4+ T cells were removed and cDNA samples were prepared and subjected to real time PCR analysis using SYBR green reagent (as described in Methods). (G) Changes of IFNγ- and IL12A mRNA levels upon repeated flagellin stimulations for 2 or 12 h each. III = three F stimulations. Data represents the mean ± SD of 3–5 separate experiments.

    Techniques Used: Expressing, Isolation, Magnetic Beads, Incubation, Recombinant, Activation Assay, Real-time Polymerase Chain Reaction, SYBR Green Assay

    22) Product Images from "Newcastle Disease Virus Vectored Bivalent Vaccine against Virulent Infectious Bursal Disease and Newcastle Disease of Chickens"

    Article Title: Newcastle Disease Virus Vectored Bivalent Vaccine against Virulent Infectious Bursal Disease and Newcastle Disease of Chickens

    Journal: Vaccines

    doi: 10.3390/vaccines5040031

    Flow cytometric analysis of CD4 + and CD8 + T cells in peripheral blood mono-nuclear cells (PBMCs) of vaccinated and unvaccinated birds at 14 ( A ) and 21 ( B ) dpi. PBMCs were collected from the birds and 1 × 10 5 cells were analyzed after addition of chicken specific Mabs conjugated with fluorescent dyes. Data represent the percentage of cells ± SEM. The values depicted in different lowercase superscripts differ significantly ( p
    Figure Legend Snippet: Flow cytometric analysis of CD4 + and CD8 + T cells in peripheral blood mono-nuclear cells (PBMCs) of vaccinated and unvaccinated birds at 14 ( A ) and 21 ( B ) dpi. PBMCs were collected from the birds and 1 × 10 5 cells were analyzed after addition of chicken specific Mabs conjugated with fluorescent dyes. Data represent the percentage of cells ± SEM. The values depicted in different lowercase superscripts differ significantly ( p

    Techniques Used: Flow Cytometry

    23) Product Images from "Increased Th17 cells and IL-17A exist in patients with B cell acute lymphoblastic leukemia and promote proliferation and resistance to daunorubicin through activation of Akt signaling"

    Article Title: Increased Th17 cells and IL-17A exist in patients with B cell acute lymphoblastic leukemia and promote proliferation and resistance to daunorubicin through activation of Akt signaling

    Journal: Journal of Translational Medicine

    doi: 10.1186/s12967-016-0894-9

    Th17 cells increase with reduced Th1 cells in freshly isolated PBMCs and BMMCs from patients with B-ALL. a PBMCs and BMMCs were separated and stimulated with PMA and ionomycin for 5 h in the presence of brefeldin A and subsequently stained with antibodies against CD3, CD8, intracellular IL-17A or IFN-γ. Flow cytometric analysis was used to determine the Th17 and Th1 cell frequencies. Representative dot plots using matching peripheral blood and bone marrow samples from B-ALL patients and healthy donors (HD) were shown. b Statistical data for frequencies of Th17 and Th1 cells within CD4 + T population were shown. c Total RNA was extracted from CD4 + T cells isolated from B-ALL patients and HDs and reverse transcribed into cDNA and subsequently determined for IL-17A and IFN-γ mRNA expression using quantitative PCR. d The frequencies of Th17 cells were significantly decreased in BM when B-ALL patients achieved complete remission (CR). e CD4 + T cells were cultured with or without Nalm-6 cells for 14 days in the presence of OKT3 plus IL-2 (300 units/ml). Then, frequencies of Th17 cells were determined after stimulation with PMA plus ionomycin
    Figure Legend Snippet: Th17 cells increase with reduced Th1 cells in freshly isolated PBMCs and BMMCs from patients with B-ALL. a PBMCs and BMMCs were separated and stimulated with PMA and ionomycin for 5 h in the presence of brefeldin A and subsequently stained with antibodies against CD3, CD8, intracellular IL-17A or IFN-γ. Flow cytometric analysis was used to determine the Th17 and Th1 cell frequencies. Representative dot plots using matching peripheral blood and bone marrow samples from B-ALL patients and healthy donors (HD) were shown. b Statistical data for frequencies of Th17 and Th1 cells within CD4 + T population were shown. c Total RNA was extracted from CD4 + T cells isolated from B-ALL patients and HDs and reverse transcribed into cDNA and subsequently determined for IL-17A and IFN-γ mRNA expression using quantitative PCR. d The frequencies of Th17 cells were significantly decreased in BM when B-ALL patients achieved complete remission (CR). e CD4 + T cells were cultured with or without Nalm-6 cells for 14 days in the presence of OKT3 plus IL-2 (300 units/ml). Then, frequencies of Th17 cells were determined after stimulation with PMA plus ionomycin

    Techniques Used: Isolation, Staining, Flow Cytometry, Expressing, Real-time Polymerase Chain Reaction, Cell Culture

    24) Product Images from "Synthetic Peptides Elicit Strong Cellular Immunity in Visceral Leishmaniasis Natural Reservoir and Contribute to Long-Lasting Polyfunctional T-Cells in BALB/c Mice"

    Article Title: Synthetic Peptides Elicit Strong Cellular Immunity in Visceral Leishmaniasis Natural Reservoir and Contribute to Long-Lasting Polyfunctional T-Cells in BALB/c Mice

    Journal: Vaccines

    doi: 10.3390/vaccines7040162

    In vitro proliferation and IFN-γ production by T-lymphocytes in peripheral blood mononuclear cells (PBMC) of L. infantum naturally infected dogs after stimuli with predicted peptides. The spots represent indexes of T-cell responses to Leishmania infantum peptide pools in PBMC of naturally infected and asymptomatic dogs ( n = 5 in gray circles). Proliferation index of CD4 + ( a ) and CD8 + ( b ) lymphocytes after peptide pool stimuli. ( c ) Index of IFN-γ producing by CD4 + and CD8 + ( d ) lymphocytes after peptide pool stimuli. Indexes were calculated based on SLA (soluble Leishmania antigen)-stimulated (SC) cultures divided by the control culture (CC). Cut-off (dashed line) for responses was calculated by the control culture indexes (cut-off = 1).
    Figure Legend Snippet: In vitro proliferation and IFN-γ production by T-lymphocytes in peripheral blood mononuclear cells (PBMC) of L. infantum naturally infected dogs after stimuli with predicted peptides. The spots represent indexes of T-cell responses to Leishmania infantum peptide pools in PBMC of naturally infected and asymptomatic dogs ( n = 5 in gray circles). Proliferation index of CD4 + ( a ) and CD8 + ( b ) lymphocytes after peptide pool stimuli. ( c ) Index of IFN-γ producing by CD4 + and CD8 + ( d ) lymphocytes after peptide pool stimuli. Indexes were calculated based on SLA (soluble Leishmania antigen)-stimulated (SC) cultures divided by the control culture (CC). Cut-off (dashed line) for responses was calculated by the control culture indexes (cut-off = 1).

    Techniques Used: In Vitro, Infection

    25) Product Images from "Potent In Vitro Activity of Citrus aurantium Essential Oil and Vitis vinifera Hydrolate Against Gut Yeast Isolates from Irritable Bowel Syndrome Patients—The Right Mix for Potential Therapeutic Use"

    Article Title: Potent In Vitro Activity of Citrus aurantium Essential Oil and Vitis vinifera Hydrolate Against Gut Yeast Isolates from Irritable Bowel Syndrome Patients—The Right Mix for Potential Therapeutic Use

    Journal: Nutrients

    doi: 10.3390/nu12051329

    Variation of the amount of cytokines IL-10 and TNF-α in the medium culture of PBMCs treated with scalar dilutions of the V. vinifera cv Italia Hy alone or in combination with EO and/or LPS. The circle-shaped indicators refer to the samples treated with the Hy only; the squares are referred to the samples treated with the Hy plus LPS, while the triangles show the samples treated with Hy plus the EO of C. aurantium var. amara plus LPS. The white rhombus is the untreated control and the black rhombus is the control treated with LPS. Letters a , b , c and d indicate samples treated with scalar dilution of the Hy (50% v / v , 25% v / v , 12.50% v / v and 0.06% v / v respectively). Letters e , f , g and h are referred to samples treated with scalar dilution of the Hy (50% v / v , 25% v / v , 12.50% v / v and 0.06% v / v respectively) and LPS (1 μgr/mL). Letters i , l , m and n are referred to PBMCs treated with a 1:100 v / v mixture of the Hy, (50% v / v , 25% v / v , 12.50% v / v and 0.06% v / v respectively) and the EO (0.50% v / v , 0.25% v / v , 0.12% v / v and 0.06% v / v respectively) plus LPS.
    Figure Legend Snippet: Variation of the amount of cytokines IL-10 and TNF-α in the medium culture of PBMCs treated with scalar dilutions of the V. vinifera cv Italia Hy alone or in combination with EO and/or LPS. The circle-shaped indicators refer to the samples treated with the Hy only; the squares are referred to the samples treated with the Hy plus LPS, while the triangles show the samples treated with Hy plus the EO of C. aurantium var. amara plus LPS. The white rhombus is the untreated control and the black rhombus is the control treated with LPS. Letters a , b , c and d indicate samples treated with scalar dilution of the Hy (50% v / v , 25% v / v , 12.50% v / v and 0.06% v / v respectively). Letters e , f , g and h are referred to samples treated with scalar dilution of the Hy (50% v / v , 25% v / v , 12.50% v / v and 0.06% v / v respectively) and LPS (1 μgr/mL). Letters i , l , m and n are referred to PBMCs treated with a 1:100 v / v mixture of the Hy, (50% v / v , 25% v / v , 12.50% v / v and 0.06% v / v respectively) and the EO (0.50% v / v , 0.25% v / v , 0.12% v / v and 0.06% v / v respectively) plus LPS.

    Techniques Used:

    26) Product Images from "Pancreatic Cancer Cell Lines Can Induce Prostaglandin E2 Production from Human Blood Mononuclear Cells"

    Article Title: Pancreatic Cancer Cell Lines Can Induce Prostaglandin E2 Production from Human Blood Mononuclear Cells

    Journal: Journal of Oncology

    doi: 10.1155/2011/741868

    Release of PGE 2 from pancreatic carcinoma cells and peripheral blood mononuclear cells. (a) PGE 2 production from PDAC monocultures of cells. Cells (AsPC-1, MiaPaCa-2, T3M-4, BxPC-3) were plated in 48-well plates at a density of 2.5 × 10 4 cells/well in 500 μ L of medium/well, and supernatants were collected at 48 hrs for PGE 2 ELISA measurement. (b) PGE 2 production from PBMCs. After purification over a Histopaque gradient mononuclear cells were plated in a 48-well plate (5 × 10 5 cells/well) either alone or onto preseeded PDAC cultures (2.5 × 10 4 cells/well). 48 hours after coculturing the cells, supernatants were collected and analysed for PGE 2 expression. PBMCs treated with LPS and ConA served as controls. All experiments were repeated in triplicate with three different healthy blood donors. *Standard deviations were calculated, and results were considered significant with P values from Student's t -test below 0.05.
    Figure Legend Snippet: Release of PGE 2 from pancreatic carcinoma cells and peripheral blood mononuclear cells. (a) PGE 2 production from PDAC monocultures of cells. Cells (AsPC-1, MiaPaCa-2, T3M-4, BxPC-3) were plated in 48-well plates at a density of 2.5 × 10 4 cells/well in 500 μ L of medium/well, and supernatants were collected at 48 hrs for PGE 2 ELISA measurement. (b) PGE 2 production from PBMCs. After purification over a Histopaque gradient mononuclear cells were plated in a 48-well plate (5 × 10 5 cells/well) either alone or onto preseeded PDAC cultures (2.5 × 10 4 cells/well). 48 hours after coculturing the cells, supernatants were collected and analysed for PGE 2 expression. PBMCs treated with LPS and ConA served as controls. All experiments were repeated in triplicate with three different healthy blood donors. *Standard deviations were calculated, and results were considered significant with P values from Student's t -test below 0.05.

    Techniques Used: Enzyme-linked Immunosorbent Assay, Purification, Expressing

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    Gradient Centrifugation:

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    Article Title: Progranulin and a Five Transmembrane Domain-Containing Receptor-like Gene Are the Key Components in Receptor Activator of Nuclear Factor κB (RANK)-dependent Formation of Multinucleated Osteoclasts *
    Article Snippet: .. Human bone marrow cells (HBMCs) and peripheral blood mononuclear cells (PBMCs) were obtained from healthy volunteers and were separated by density gradient centrifugation using Ficoll-Histopaque (Sigma-Aldrich). .. These cells were cultured for 7 days in the presence of M-CSF (100 ng/ml).

    In Vitro:

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    Isolation:

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    Article Title: Novel oligodeoxynucleotide agonists of TLR9 containing N3-Me-dC or N1-Me-dG modifications
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    Article Title: Fucoidan inhibits CCL22 production through NF-κB pathway in M2 macrophages: a potential therapeutic strategy for cancer
    Article Snippet: .. Preparation and isolation of PBMC and cell subtypes Human peripheral blood mononuclear cells (PBMCs) were isolated from leukocyte-enriched buffy coats of healthy donors by Ficoll (Sigma-Aldrich) density gradient centrifugation. ..

    Article Title: Cyclosporin A-Sensitive Transcription Factor Egr-3 Regulates Fas Ligand Expression
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    Cell Culture:

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    Activity Assay:

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    Recombinant:

    Article Title: Cyclosporin A-Sensitive Transcription Factor Egr-3 Regulates Fas Ligand Expression
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  • 89
    Millipore culture pbmcs
    Features of ssRNA40-induced LILRA3 expression. a Kinetics of LILRA3 expression was compared to other ssRNA40 inducible cytokines. <t>PBMCs</t> from two donors were stimulated with ssRNA40 or ssRNA41 and the RNA was harvested at various time points. qPCR for LILRA3 , TNF , IFNb , IFNg , and IL1b expression, normalized to RPLP0 and relatively quantified to calibrator cDNA (pooled cDNA from 10. b Effect of cytokine inhibition on LILRA3 expression. PBMCs from two donors were stimulated overnight with ssRNA40 together with neutralizing agents anakinra, certolizumab, tocilizumab and IFN-γ neutralizing antibodies and analysed for LILRA3 and IL6 expression by qPCR. Repeated measures ANOVA was used to calculate difference of ssRNA40+ neutralization to ssRNA40 alone and results displayed as mean ± SD
    Culture Pbmcs, supplied by Millipore, used in various techniques. Bioz Stars score: 89/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore pbmc specimens against sd1 lps
    Fecal s-IgA antibody titers to <t>SD1-LPS</t> in group1 (cross symbol), group 2 (open square symbol), and group 3 (closed round symbol) monkeys, expressed as MN with MN ± SE, after challenge on day 00 and re-challenge on day 31 with SD11617 strain.
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    Millipore human pbmcs
    Imatinib inhibits TNF-α production in human myeloid cells. Human <t>PBMCs,</t> monocytes, or macrophages were exposed to either saline or imatinib (1 h, 1 μM) followed by 3-h stimulation with 100 ng/ml <t>LPS.</t> ( A ) LPS-induced TNF-α levels in supernatants of all three cell types as determined by ELISA ( n = 5; * , P ≤ 0.01). ( B and C ) Imatinib dose-dependently inhibits LPS-induced TNF-α protein ( B ) ( n = 5; * , P ≤ 0.01) and TNF-α mRNA expression ( C ) ( n = 3; * , P ≤ 0.01) in human PBMCs.
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    Millipore host pbmcs
    Effects of the various concentrations of <t>rHcMTF-12</t> on <t>PBMC</t> migration in vitro . Cells were treated with control buffer, pET32a protein and multiple concentrations of rHcMTF-12. The migration percentage was determined randomly. The difference between the mean values was calculated using ANOVA. PBMC used for all replicates of distinct treatments in each experimental repetition were derived from the same goat. Data are representative of three independent experiments. * P
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    Features of ssRNA40-induced LILRA3 expression. a Kinetics of LILRA3 expression was compared to other ssRNA40 inducible cytokines. PBMCs from two donors were stimulated with ssRNA40 or ssRNA41 and the RNA was harvested at various time points. qPCR for LILRA3 , TNF , IFNb , IFNg , and IL1b expression, normalized to RPLP0 and relatively quantified to calibrator cDNA (pooled cDNA from 10. b Effect of cytokine inhibition on LILRA3 expression. PBMCs from two donors were stimulated overnight with ssRNA40 together with neutralizing agents anakinra, certolizumab, tocilizumab and IFN-γ neutralizing antibodies and analysed for LILRA3 and IL6 expression by qPCR. Repeated measures ANOVA was used to calculate difference of ssRNA40+ neutralization to ssRNA40 alone and results displayed as mean ± SD

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

    doi: 10.1186/s12977-016-0248-y

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

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

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

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

    Journal: Retrovirology

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

    doi: 10.1186/s12977-016-0248-y

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

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

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

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

    Journal: Retrovirology

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

    doi: 10.1186/s12977-016-0248-y

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

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

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

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

    Journal: Apmis

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

    doi: 10.1111/apm.12168

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

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

    Techniques:

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

    Journal: Apmis

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

    doi: 10.1111/apm.12168

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

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

    Techniques:

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

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

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

    doi: 10.1073/pnas.0501758102

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

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

    Techniques: Enzyme-linked Immunosorbent Assay, Expressing

    Effects of the various concentrations of rHcMTF-12 on PBMC migration in vitro . Cells were treated with control buffer, pET32a protein and multiple concentrations of rHcMTF-12. The migration percentage was determined randomly. The difference between the mean values was calculated using ANOVA. PBMC used for all replicates of distinct treatments in each experimental repetition were derived from the same goat. Data are representative of three independent experiments. * P

    Journal: Parasites & Vectors

    Article Title: Identification of a novel methyltransferase-type 12 protein from Haemonchus contortus and its effects on functions of goat PBMCs

    doi: 10.1186/s13071-020-04028-y

    Figure Lengend Snippet: Effects of the various concentrations of rHcMTF-12 on PBMC migration in vitro . Cells were treated with control buffer, pET32a protein and multiple concentrations of rHcMTF-12. The migration percentage was determined randomly. The difference between the mean values was calculated using ANOVA. PBMC used for all replicates of distinct treatments in each experimental repetition were derived from the same goat. Data are representative of three independent experiments. * P

    Article Snippet: rHcMTF-12 promoted migration efficiency of PBMCs The change in migratory efficiency of host PBMCs in response to rHcMTF-12 was assessed by the percentile of shifted PBMCs through the Millipore polycarbonate membrane into the lower chamber.

    Techniques: Migration, In Vitro, Derivative Assay

    Relative expression of multiple cytokines in goat PBMCs stimulated by rHcMTF-12. Cells were incubated with rHcMTF-12 for 48 h, and the mRNAs encoding IL-2, IL-4, IL-6, IL-10, TGF-β1 and IFN-γ were quantified by real-time PCR. The significant level was set at * P

    Journal: Parasites & Vectors

    Article Title: Identification of a novel methyltransferase-type 12 protein from Haemonchus contortus and its effects on functions of goat PBMCs

    doi: 10.1186/s13071-020-04028-y

    Figure Lengend Snippet: Relative expression of multiple cytokines in goat PBMCs stimulated by rHcMTF-12. Cells were incubated with rHcMTF-12 for 48 h, and the mRNAs encoding IL-2, IL-4, IL-6, IL-10, TGF-β1 and IFN-γ were quantified by real-time PCR. The significant level was set at * P

    Article Snippet: rHcMTF-12 promoted migration efficiency of PBMCs The change in migratory efficiency of host PBMCs in response to rHcMTF-12 was assessed by the percentile of shifted PBMCs through the Millipore polycarbonate membrane into the lower chamber.

    Techniques: Expressing, Incubation, Real-time Polymerase Chain Reaction

    Decreased level of cell proliferation by rHcMTF-12 in vitro . Cells were simulated with ConA and treated with control buffer (PBS), pET32a protein and various concentrations of rHcMTF-12 protein for 72 h. A proliferation test was conducted by CCK-8 and values were measured at 450 nm wavelength (OD 450 ) using microplate spectrophotometer. Cell proliferation index was calculated by considering the OD 450 values in the controls as 100%. PBMC used for all replicates of distinct treatments in each experimental repetition were derived from the same goat. The data are expressed as the mean ± standard error (SE) of three independent experiments. ** P

    Journal: Parasites & Vectors

    Article Title: Identification of a novel methyltransferase-type 12 protein from Haemonchus contortus and its effects on functions of goat PBMCs

    doi: 10.1186/s13071-020-04028-y

    Figure Lengend Snippet: Decreased level of cell proliferation by rHcMTF-12 in vitro . Cells were simulated with ConA and treated with control buffer (PBS), pET32a protein and various concentrations of rHcMTF-12 protein for 72 h. A proliferation test was conducted by CCK-8 and values were measured at 450 nm wavelength (OD 450 ) using microplate spectrophotometer. Cell proliferation index was calculated by considering the OD 450 values in the controls as 100%. PBMC used for all replicates of distinct treatments in each experimental repetition were derived from the same goat. The data are expressed as the mean ± standard error (SE) of three independent experiments. ** P

    Article Snippet: rHcMTF-12 promoted migration efficiency of PBMCs The change in migratory efficiency of host PBMCs in response to rHcMTF-12 was assessed by the percentile of shifted PBMCs through the Millipore polycarbonate membrane into the lower chamber.

    Techniques: In Vitro, CCK-8 Assay, Spectrophotometry, Derivative Assay

    Binding of rHcMTF-12 protein with goat PBMCs. The cells were incubated with rat sera anti-rHcMTF-12 IgG, negative rat IgG or anti-pET32a protein IgG as the primary antibody followed by Cy3-labeled goat anti-rat IgG as the secondary antibody. Red fluorescence on surface of cells showed target protein staining (Cy3) and cell nuclei were visualized by DAPI (blue). No fluorescence was observed in the PBS control or pET32 control groups. Scale-bars : 10 μm

    Journal: Parasites & Vectors

    Article Title: Identification of a novel methyltransferase-type 12 protein from Haemonchus contortus and its effects on functions of goat PBMCs

    doi: 10.1186/s13071-020-04028-y

    Figure Lengend Snippet: Binding of rHcMTF-12 protein with goat PBMCs. The cells were incubated with rat sera anti-rHcMTF-12 IgG, negative rat IgG or anti-pET32a protein IgG as the primary antibody followed by Cy3-labeled goat anti-rat IgG as the secondary antibody. Red fluorescence on surface of cells showed target protein staining (Cy3) and cell nuclei were visualized by DAPI (blue). No fluorescence was observed in the PBS control or pET32 control groups. Scale-bars : 10 μm

    Article Snippet: rHcMTF-12 promoted migration efficiency of PBMCs The change in migratory efficiency of host PBMCs in response to rHcMTF-12 was assessed by the percentile of shifted PBMCs through the Millipore polycarbonate membrane into the lower chamber.

    Techniques: Binding Assay, Incubation, Labeling, Fluorescence, Staining