peripheral blood mononuclear cells pbmcs  (Lonza)


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  • 99
    Name:
    Human Peripheral Blood Mononuclear Cells
    Description:
    Mononuclear cells from human peripheral blood cryopreserved 10 million cells
    Catalog Number:
    4w-270
    Price:
    None
    Category:
    Primary and Stem Cells
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    Structured Review

    Lonza peripheral blood mononuclear cells pbmcs
    ShK-186 inhibits antineutrophil cytoplasmic antibodies targeting proteinase 3 (PR3-ANCA) production in vitro . (A) IgG production after peripheral blood mononuclear cell <t>(PBMC)</t> stimulation with CpG, B cell-activating factor (BAFF), and interleukin (IL)-21 from 5 healthy controls (HCs) (open circles) and 13 granulomatosis with polyangiitis <t>(GPA)</t> patients (gray circles) in the presence and absence of 1 nM ShK-186. (B) PR3-ANCA production after PBMC stimulation with CpG, BAFF, and IL-21 from 13 GPA patients in the presence and absence of 1 nM ShK-186 (* p
    Mononuclear cells from human peripheral blood cryopreserved 10 million cells
    https://www.bioz.com/result/peripheral blood mononuclear cells pbmcs/product/Lonza
    Average 99 stars, based on 20 article reviews
    Price from $9.99 to $1999.99
    peripheral blood mononuclear cells pbmcs - by Bioz Stars, 2020-09
    99/100 stars

    Images

    1) Product Images from "Kv1.3 Channel Blockade Modulates the Effector Function of B Cells in Granulomatosis with Polyangiitis"

    Article Title: Kv1.3 Channel Blockade Modulates the Effector Function of B Cells in Granulomatosis with Polyangiitis

    Journal: Frontiers in Immunology

    doi: 10.3389/fimmu.2017.01205

    ShK-186 inhibits antineutrophil cytoplasmic antibodies targeting proteinase 3 (PR3-ANCA) production in vitro . (A) IgG production after peripheral blood mononuclear cell (PBMC) stimulation with CpG, B cell-activating factor (BAFF), and interleukin (IL)-21 from 5 healthy controls (HCs) (open circles) and 13 granulomatosis with polyangiitis (GPA) patients (gray circles) in the presence and absence of 1 nM ShK-186. (B) PR3-ANCA production after PBMC stimulation with CpG, BAFF, and IL-21 from 13 GPA patients in the presence and absence of 1 nM ShK-186 (* p
    Figure Legend Snippet: ShK-186 inhibits antineutrophil cytoplasmic antibodies targeting proteinase 3 (PR3-ANCA) production in vitro . (A) IgG production after peripheral blood mononuclear cell (PBMC) stimulation with CpG, B cell-activating factor (BAFF), and interleukin (IL)-21 from 5 healthy controls (HCs) (open circles) and 13 granulomatosis with polyangiitis (GPA) patients (gray circles) in the presence and absence of 1 nM ShK-186. (B) PR3-ANCA production after PBMC stimulation with CpG, BAFF, and IL-21 from 13 GPA patients in the presence and absence of 1 nM ShK-186 (* p

    Techniques Used: In Vitro

    Related Articles

    Centrifugation:

    Article Title: Porcupine Is Not Required for the Production of the Majority of Wnts from Primary Human Astrocytes and CD8+ T Cells
    Article Snippet: .. Culture and Activation of Human Peripheral Blood Mononuclear Cells (PBMCs) and Isolation of CD8+ T cells PBMCs were isolated from venous blood of healthy donors using lymphocyte separation medium (Lonza Biologics, Portsmouth, NH) and density centrifugation. .. PBMCs were then cultured at 1×106 cells mL of RPMI 1640, 10% heat-inactivated fetal bovine serum (FBS), 200 mM L-glutamine, and 1% penicillin/streptomycin (Lonza Biologics, Portsmouth, NH).

    In Vitro:

    Article Title: Kv1.3 Channel Blockade Modulates the Effector Function of B Cells in Granulomatosis with Polyangiitis
    Article Snippet: .. Briefly, peripheral blood mononuclear cells (PBMCs) were isolated from GPA patients, who produce ANCA upon in vitro induction , and stored in RPMI 1640 (Lonza, Basel, Switzerland) supplemented with 50 µg/mL gentamycin (GIBCO, Life Technologies, Grand Island, NY, USA), 10% fetal calf serum (FCS, Lonza), and 10% dimethyl sulfoxide (DMSO). ..

    Co-Culture Assay:

    Article Title: High expression of IDO1 and TGF-β1 during recurrence and post infection clearance with Chlamydia trachomatis, are independent of host IFN-γ response
    Article Snippet: .. Experiments were conducted in 48-well plates, where co-culture of 40,000 ECC1 cells/well and 8 × 105 female human PBMCs (Lonza, Australia) cells/well were seeded with RPMI-1640 medium (Sigma-Aldrich, Australia) containing 10% heat inactivated FCS (Life Technologies, Australia), 120 μg/ml streptomycin (Sigma-Aldrich, Australia), 50 μg/ml Gentamycin (Gibco, Australia), 37 °C, 5% CO2 . .. In addition, control plates with either ECC1 cells alone or human PBMCs alone were used.

    Isolation:

    Article Title: P2X7 Receptor Induces Tumor Necrosis Factor-α Converting Enzyme Activation and Release to Boost TNF-α Production
    Article Snippet: .. Human peripheral blood mononuclear cells were isolated following standard procedure ( ) and cultured for 16 h in RPMI 1640 medium (Lonza) with 10% of FCS, 2 mM Glutamax, and 100 U/ml penicillin–streptomycin (Life Technologies). .. After monocyte adherence, cells were washed and primed for 4 h with LPS (10 ng/ml), and then cells were washed or not with physiological buffer and incubated in the same buffer at 37°C with 3 mM of ATP for 20 min.

    Article Title: Porcupine Is Not Required for the Production of the Majority of Wnts from Primary Human Astrocytes and CD8+ T Cells
    Article Snippet: .. Culture and Activation of Human Peripheral Blood Mononuclear Cells (PBMCs) and Isolation of CD8+ T cells PBMCs were isolated from venous blood of healthy donors using lymphocyte separation medium (Lonza Biologics, Portsmouth, NH) and density centrifugation. .. PBMCs were then cultured at 1×106 cells mL of RPMI 1640, 10% heat-inactivated fetal bovine serum (FBS), 200 mM L-glutamine, and 1% penicillin/streptomycin (Lonza Biologics, Portsmouth, NH).

    Article Title: Kv1.3 Channel Blockade Modulates the Effector Function of B Cells in Granulomatosis with Polyangiitis
    Article Snippet: .. Briefly, peripheral blood mononuclear cells (PBMCs) were isolated from GPA patients, who produce ANCA upon in vitro induction , and stored in RPMI 1640 (Lonza, Basel, Switzerland) supplemented with 50 µg/mL gentamycin (GIBCO, Life Technologies, Grand Island, NY, USA), 10% fetal calf serum (FCS, Lonza), and 10% dimethyl sulfoxide (DMSO). ..

    Cell Culture:

    Article Title: P2X7 Receptor Induces Tumor Necrosis Factor-α Converting Enzyme Activation and Release to Boost TNF-α Production
    Article Snippet: .. Human peripheral blood mononuclear cells were isolated following standard procedure ( ) and cultured for 16 h in RPMI 1640 medium (Lonza) with 10% of FCS, 2 mM Glutamax, and 100 U/ml penicillin–streptomycin (Life Technologies). .. After monocyte adherence, cells were washed and primed for 4 h with LPS (10 ng/ml), and then cells were washed or not with physiological buffer and incubated in the same buffer at 37°C with 3 mM of ATP for 20 min.

    Activation Assay:

    Article Title: Porcupine Is Not Required for the Production of the Majority of Wnts from Primary Human Astrocytes and CD8+ T Cells
    Article Snippet: .. Culture and Activation of Human Peripheral Blood Mononuclear Cells (PBMCs) and Isolation of CD8+ T cells PBMCs were isolated from venous blood of healthy donors using lymphocyte separation medium (Lonza Biologics, Portsmouth, NH) and density centrifugation. .. PBMCs were then cultured at 1×106 cells mL of RPMI 1640, 10% heat-inactivated fetal bovine serum (FBS), 200 mM L-glutamine, and 1% penicillin/streptomycin (Lonza Biologics, Portsmouth, NH).

    other:

    Article Title: High expression of IDO1 and TGF-β1 during recurrence and post infection clearance with Chlamydia trachomatis, are independent of host IFN-γ response
    Article Snippet: In addition, control plates with either ECC1 cells alone or human PBMCs alone were used.

    Infection:

    Article Title: High expression of IDO1 and TGF-β1 during recurrence and post infection clearance with Chlamydia trachomatis, are independent of host IFN-γ response
    Article Snippet: .. In addition, infection of human PBMCs alone with live C. trachomatis , without the azithromycin treatment, resulted in significantly reduced, yet apparent infectivity (p < 0.0001). .. The number of chlamydial IFUs measured from ECC1 and PBMCs co-culture was significantly reduced in comparison to infected ECC1 culture alone (p < 0.0001) by almost two logs.

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    Lonza cd8 t cells pbmcs
    IWP-2 does not inhibit Wnt production from primary human CD8+ T cells. CD8+ T cells were isolated from <t>PBMCs</t> from healthy donors by negative selection and subsequently activated with 1 μg each anti-CD3/anti-CD28 then propagated in presence of 100 units/ml IL-2 for three days. On the third day supernatant was collected and 25 μl was analyzed by western blot for presence of Wnt 1 (A), 3 (B), 6 (C), 7a (D), 10a (E), and 16a (F).
    Cd8 T Cells Pbmcs, supplied by Lonza, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cd8 t cells pbmcs/product/Lonza
    Average 86 stars, based on 1 article reviews
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    85
    Lonza glycolipid pulsed autologous pbmcs
    Cytokine levels in supernatant of co-culture of human <t>iNKT</t> cells and glycolipid-loaded irradiated <t>PBMC</t>
    Glycolipid Pulsed Autologous Pbmcs, supplied by Lonza, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 85 stars, based on 1 article reviews
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    pbmc  (Lonza)
    93
    Lonza pbmc
    Functional analysis of MAIT cells in individuals of different ages. <t>PBMC</t> or CBMC from the US cohort were incubated overnight with M. smegmatis -infected A549 cells or uninfected A549 cells and then stained with the MR1-5-OP-RU or MR1/6FP tetramers, followed by staining with a live/dead discriminator and antibodies to TCRγδ, CD3, CD4, CD8, TRAV1-2, CD26 and CD161. <t>ICS</t> was then performed and the cells stained for TNF. A . Dot plots showing representative co-staining of live, TCRγδ - CD3 + TRAV1-2 + CD26 + CD161 + cells with MR1-5-OP-RU and TNF in M. smegmatis stimulated or unstimulated samples. The gating strategy is shown in Figure S3. Examples of the TNF response in a neonate, infant and adult are shown. B . Frequencies of functional, phenotypically-defined (CD3 + TRAV1-2 + CD26 + CD161 + ) MAIT cells that are MR1-5-OP-RU + or MR1-5-OP-RU - as a proportion of CD3 + TCRγδ - T cells. Horizontal lines depict the median and the error bars the 95% confidence interval. Wilcoxon-rank sum was used to test differences within the same cohort.
    Pbmc, supplied by Lonza, used in various techniques. Bioz Stars score: 93/100, based on 168 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pbmc/product/Lonza
    Average 93 stars, based on 168 article reviews
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    pbmc - by Bioz Stars, 2020-09
    93/100 stars
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    Image Search Results


    IWP-2 does not inhibit Wnt production from primary human CD8+ T cells. CD8+ T cells were isolated from PBMCs from healthy donors by negative selection and subsequently activated with 1 μg each anti-CD3/anti-CD28 then propagated in presence of 100 units/ml IL-2 for three days. On the third day supernatant was collected and 25 μl was analyzed by western blot for presence of Wnt 1 (A), 3 (B), 6 (C), 7a (D), 10a (E), and 16a (F).

    Journal: PLoS ONE

    Article Title: Porcupine Is Not Required for the Production of the Majority of Wnts from Primary Human Astrocytes and CD8+ T Cells

    doi: 10.1371/journal.pone.0092159

    Figure Lengend Snippet: IWP-2 does not inhibit Wnt production from primary human CD8+ T cells. CD8+ T cells were isolated from PBMCs from healthy donors by negative selection and subsequently activated with 1 μg each anti-CD3/anti-CD28 then propagated in presence of 100 units/ml IL-2 for three days. On the third day supernatant was collected and 25 μl was analyzed by western blot for presence of Wnt 1 (A), 3 (B), 6 (C), 7a (D), 10a (E), and 16a (F).

    Article Snippet: Culture and Activation of Human Peripheral Blood Mononuclear Cells (PBMCs) and Isolation of CD8+ T cells PBMCs were isolated from venous blood of healthy donors using lymphocyte separation medium (Lonza Biologics, Portsmouth, NH) and density centrifugation.

    Techniques: Isolation, Selection, Western Blot

    Cytokine levels in supernatant of co-culture of human iNKT cells and glycolipid-loaded irradiated PBMC

    Journal: Journal of immunology (Baltimore, Md. : 1950)

    Article Title: Enhanced TCR footprint by a novel glycolipid increases NKT dependent tumor protection

    doi: 10.4049/jimmunol.1203134

    Figure Lengend Snippet: Cytokine levels in supernatant of co-culture of human iNKT cells and glycolipid-loaded irradiated PBMC

    Article Snippet: Subsequently, 5×104 iNKT cells were stimulated with 105 glycolipid pulsed autologous PBMCs in RPMI 1640 media supplemented with 10% human AB serum (Lonza), 1% sodium pyruvate, 1% nonessential amino acids and 1% penicillin/streptomycin (all from Invitrogen).

    Techniques: Co-Culture Assay, Irradiation

    Functional analysis of MAIT cells in individuals of different ages. PBMC or CBMC from the US cohort were incubated overnight with M. smegmatis -infected A549 cells or uninfected A549 cells and then stained with the MR1-5-OP-RU or MR1/6FP tetramers, followed by staining with a live/dead discriminator and antibodies to TCRγδ, CD3, CD4, CD8, TRAV1-2, CD26 and CD161. ICS was then performed and the cells stained for TNF. A . Dot plots showing representative co-staining of live, TCRγδ - CD3 + TRAV1-2 + CD26 + CD161 + cells with MR1-5-OP-RU and TNF in M. smegmatis stimulated or unstimulated samples. The gating strategy is shown in Figure S3. Examples of the TNF response in a neonate, infant and adult are shown. B . Frequencies of functional, phenotypically-defined (CD3 + TRAV1-2 + CD26 + CD161 + ) MAIT cells that are MR1-5-OP-RU + or MR1-5-OP-RU - as a proportion of CD3 + TCRγδ - T cells. Horizontal lines depict the median and the error bars the 95% confidence interval. Wilcoxon-rank sum was used to test differences within the same cohort.

    Journal: bioRxiv

    Article Title: Postnatal Expansion, Maturation and Functionality of MR1T Cells in Humans

    doi: 10.1101/2019.12.20.882746

    Figure Lengend Snippet: Functional analysis of MAIT cells in individuals of different ages. PBMC or CBMC from the US cohort were incubated overnight with M. smegmatis -infected A549 cells or uninfected A549 cells and then stained with the MR1-5-OP-RU or MR1/6FP tetramers, followed by staining with a live/dead discriminator and antibodies to TCRγδ, CD3, CD4, CD8, TRAV1-2, CD26 and CD161. ICS was then performed and the cells stained for TNF. A . Dot plots showing representative co-staining of live, TCRγδ - CD3 + TRAV1-2 + CD26 + CD161 + cells with MR1-5-OP-RU and TNF in M. smegmatis stimulated or unstimulated samples. The gating strategy is shown in Figure S3. Examples of the TNF response in a neonate, infant and adult are shown. B . Frequencies of functional, phenotypically-defined (CD3 + TRAV1-2 + CD26 + CD161 + ) MAIT cells that are MR1-5-OP-RU + or MR1-5-OP-RU - as a proportion of CD3 + TCRγδ - T cells. Horizontal lines depict the median and the error bars the 95% confidence interval. Wilcoxon-rank sum was used to test differences within the same cohort.

    Article Snippet: Flow cytometry assays To perform ICS, cryopreserved PBMC were thawed in the presence of DNAse, resuspended, in 10% heat inactivated human serum with RPMI (Lonza) at a concentration of 2 × 10^7/ml.

    Techniques: Functional Assay, Incubation, Infection, Staining

    Phenotypic analysis of functional MR1T cells at different ages. PBMC or CBMC from the US cohort were incubated overnight with M. smegmatis -infected A549 cells, uninfected A549 cells, or uninfected A549 cells and PMA/ionomycin. All cells were then stained with the MR1-5-OP-RU or MR1-6FP tetramers, followed by a live/dead discriminator and antibodies to TCRγδ, CD3, CD4, CD8, TRAV1-2, CD26 and CD161. ICS was then performed and the cells stained for TNF. Live, TCRγδ - CD3 + MR1-5-OP-RU + cells were gated and the TNF + percentage determined in both the M. smegmatis stimulated and unstimulated conditions. The gating strategy is shown in Figure S3. A . Dot plots showing TNF expression by live, TCRγδ - MR1-5-OP-RU + CD3 + cells in the same representative neonate, infant and adult as Figure 2A . B . Background subtracted frequencies of TNF + MR1-5-OP-RU + CD3 + cells in response to M. smegmatis -infected A549 cells as a percentage of total MR1/5-OP-RU + CD3 + cells are shown. Mann-Whitney u-tests were used to test differences between groups. C . Background subtracted frequencies of TNF + cells to PMA/ionomycin among different T cell subpopulations, 1) MR1T cells (CD3 + TCRγδ - MR1-5-OP-RU + cells); 2) γδ T cells (CD3 + TCRγδ + MR1-5-OP-RU - cells); 3) CD8 + T cells (CD3 + TCRγδ - MR1-5-OP-RU - CD8 + cells); and 4) CD4 + T cells (CD3 + TCRγδ - MR1-5-OP-RU - CD4 + cells). Wilcoxon-rank sum was used to test differences within the same cohort. D . and E . Frequencies of TNF + TRAV1-2 + MR1-5-OP-RU + CD3 + and TNF + TRAV1-2 - MR1-5-OP-RU + CD3 + cells in response to M. smegmatis infected A549 cells ( D ) or PMA/ionomycin stimulation ( E ) minus the background frequencies of TNF + TRAV1-2 + /- MR1-5-OP-RU + CD3 + cells. Wilcoxon-rank sum was used to test differences within the same cohort.

    Journal: bioRxiv

    Article Title: Postnatal Expansion, Maturation and Functionality of MR1T Cells in Humans

    doi: 10.1101/2019.12.20.882746

    Figure Lengend Snippet: Phenotypic analysis of functional MR1T cells at different ages. PBMC or CBMC from the US cohort were incubated overnight with M. smegmatis -infected A549 cells, uninfected A549 cells, or uninfected A549 cells and PMA/ionomycin. All cells were then stained with the MR1-5-OP-RU or MR1-6FP tetramers, followed by a live/dead discriminator and antibodies to TCRγδ, CD3, CD4, CD8, TRAV1-2, CD26 and CD161. ICS was then performed and the cells stained for TNF. Live, TCRγδ - CD3 + MR1-5-OP-RU + cells were gated and the TNF + percentage determined in both the M. smegmatis stimulated and unstimulated conditions. The gating strategy is shown in Figure S3. A . Dot plots showing TNF expression by live, TCRγδ - MR1-5-OP-RU + CD3 + cells in the same representative neonate, infant and adult as Figure 2A . B . Background subtracted frequencies of TNF + MR1-5-OP-RU + CD3 + cells in response to M. smegmatis -infected A549 cells as a percentage of total MR1/5-OP-RU + CD3 + cells are shown. Mann-Whitney u-tests were used to test differences between groups. C . Background subtracted frequencies of TNF + cells to PMA/ionomycin among different T cell subpopulations, 1) MR1T cells (CD3 + TCRγδ - MR1-5-OP-RU + cells); 2) γδ T cells (CD3 + TCRγδ + MR1-5-OP-RU - cells); 3) CD8 + T cells (CD3 + TCRγδ - MR1-5-OP-RU - CD8 + cells); and 4) CD4 + T cells (CD3 + TCRγδ - MR1-5-OP-RU - CD4 + cells). Wilcoxon-rank sum was used to test differences within the same cohort. D . and E . Frequencies of TNF + TRAV1-2 + MR1-5-OP-RU + CD3 + and TNF + TRAV1-2 - MR1-5-OP-RU + CD3 + cells in response to M. smegmatis infected A549 cells ( D ) or PMA/ionomycin stimulation ( E ) minus the background frequencies of TNF + TRAV1-2 + /- MR1-5-OP-RU + CD3 + cells. Wilcoxon-rank sum was used to test differences within the same cohort.

    Article Snippet: Flow cytometry assays To perform ICS, cryopreserved PBMC were thawed in the presence of DNAse, resuspended, in 10% heat inactivated human serum with RPMI (Lonza) at a concentration of 2 × 10^7/ml.

    Techniques: Functional Assay, Incubation, Infection, Staining, Expressing, MANN-WHITNEY

    Frequencies of tetramer-defined MR1T cell and phenotypically defined MAIT cell populations in peripheral blood from individuals of different ages. PBMC or CBMC were stained with a live/dead discriminator, antibodies to CD3, CD4, CD8, TRAV1-2, CD26, CD161, and either the MR1-5-OP-RU or MR1-6FP tetramers. Live, CD3 + lymphocytes were gated and the frequencies of MR1-5-OP-RU + or TRAV1-2 + CD26 + CD161 + cells as a percentage of CD3 + lymphocytes were determined (gating strategy in Figure S2). A . Flow cytometry plots showing CD3 + T cell staining with MR1-5-OP-RU tetramer, MR1-6FP tetramer, TRAV1-2 or CD26/CD161 of samples from a representative US adult, infant and neonate. B . Frequencies of tetramer-defined (CD3 + MR1-5-OP-RU + ) and phenotypically-defined (CD3 + TRAV1-2 + CD26 + CD161 + ) MAIT cells in neonates, 10 week-old infants and adolescents from South Africa, neonates, 12 month-old infants and adults from the United States and infants (0-2 years old), children (2-5 years old) and adults from Uganda (all cohorts, n =10). Mann-Whitney u-tests were used to test differences between groups. Horizontal lines depict the median and the error bars the 95% confidence interval. C . Relative proportions of median phenotypically-defined (CD3 + TRAV1-2 + CD26 + CD161 + ) MAIT cells that are MR1-5-OP-RU + (Blue) and MR1-5-OP-RU - (Red shades) for each age group at each site. In each doughnut chart the MR1-5-OP-RU - population is further divided into CD8 + , CD8 + CD4 + , CD4 + , and CD8 - CD4 - populations.

    Journal: bioRxiv

    Article Title: Postnatal Expansion, Maturation and Functionality of MR1T Cells in Humans

    doi: 10.1101/2019.12.20.882746

    Figure Lengend Snippet: Frequencies of tetramer-defined MR1T cell and phenotypically defined MAIT cell populations in peripheral blood from individuals of different ages. PBMC or CBMC were stained with a live/dead discriminator, antibodies to CD3, CD4, CD8, TRAV1-2, CD26, CD161, and either the MR1-5-OP-RU or MR1-6FP tetramers. Live, CD3 + lymphocytes were gated and the frequencies of MR1-5-OP-RU + or TRAV1-2 + CD26 + CD161 + cells as a percentage of CD3 + lymphocytes were determined (gating strategy in Figure S2). A . Flow cytometry plots showing CD3 + T cell staining with MR1-5-OP-RU tetramer, MR1-6FP tetramer, TRAV1-2 or CD26/CD161 of samples from a representative US adult, infant and neonate. B . Frequencies of tetramer-defined (CD3 + MR1-5-OP-RU + ) and phenotypically-defined (CD3 + TRAV1-2 + CD26 + CD161 + ) MAIT cells in neonates, 10 week-old infants and adolescents from South Africa, neonates, 12 month-old infants and adults from the United States and infants (0-2 years old), children (2-5 years old) and adults from Uganda (all cohorts, n =10). Mann-Whitney u-tests were used to test differences between groups. Horizontal lines depict the median and the error bars the 95% confidence interval. C . Relative proportions of median phenotypically-defined (CD3 + TRAV1-2 + CD26 + CD161 + ) MAIT cells that are MR1-5-OP-RU + (Blue) and MR1-5-OP-RU - (Red shades) for each age group at each site. In each doughnut chart the MR1-5-OP-RU - population is further divided into CD8 + , CD8 + CD4 + , CD4 + , and CD8 - CD4 - populations.

    Article Snippet: Flow cytometry assays To perform ICS, cryopreserved PBMC were thawed in the presence of DNAse, resuspended, in 10% heat inactivated human serum with RPMI (Lonza) at a concentration of 2 × 10^7/ml.

    Techniques: Staining, Flow Cytometry, MANN-WHITNEY

    EHV-4 downregulates cell surface adhesion molecules on peripheral blood mononuclear cells (PBMC). ( A ) surface expression of very late antigen-4 (VLA-4) and lymphocyte function-associated antigen-1 (LFA-1) on the surface of mock-infected PBMC and virus infection ( black line: EHV-1; dotted line: EHV-4) levels of PBMC were determined by flow cytometry. ( B and C ) Equine PBMC were either mock-infected or infected with EHV-4 or EHV-1 and subjected to flow cytometric analysis. Histograms and bars of VLA-4 ( B ) and LFA-1 ( C ) expression after detection with the relevant antibodies are shown. Viable cells (10,000) were analyzed for each sample. Grey filled: mock-infected cells; dotted line: mock-infected cells with the relevant antibody; black line: EHV-4-infected cells; grey line: EHV-1-infected cells. For the bars, the expression level of adhesion molecules (VLA-4 or LFA-1) was set to 100%. All data represent the mean ± SD of three independent experiments. A significant downregulation (one-way ANOVA; p

    Journal: Viruses

    Article Title: The Role of the Equine Herpesvirus Type 1 (EHV-1) US3-Encoded Protein Kinase in Actin Reorganization and Nuclear Egress

    doi: 10.3390/v8100275

    Figure Lengend Snippet: EHV-4 downregulates cell surface adhesion molecules on peripheral blood mononuclear cells (PBMC). ( A ) surface expression of very late antigen-4 (VLA-4) and lymphocyte function-associated antigen-1 (LFA-1) on the surface of mock-infected PBMC and virus infection ( black line: EHV-1; dotted line: EHV-4) levels of PBMC were determined by flow cytometry. ( B and C ) Equine PBMC were either mock-infected or infected with EHV-4 or EHV-1 and subjected to flow cytometric analysis. Histograms and bars of VLA-4 ( B ) and LFA-1 ( C ) expression after detection with the relevant antibodies are shown. Viable cells (10,000) were analyzed for each sample. Grey filled: mock-infected cells; dotted line: mock-infected cells with the relevant antibody; black line: EHV-4-infected cells; grey line: EHV-1-infected cells. For the bars, the expression level of adhesion molecules (VLA-4 or LFA-1) was set to 100%. All data represent the mean ± SD of three independent experiments. A significant downregulation (one-way ANOVA; p

    Article Snippet: PBMCs were infected with either parental (EHV-1 or EHV-4) or mutant (EHV-1∆US3) viruses (MOI = 1) or transfected with different expression vectors (pcDNA3, pcDNA3-US3_1 or pcDNA3-US3_4; transfection efficiency was around 40%) using Nucleofector™ (Lonza, Köln, Germany) [ ].

    Techniques: Expressing, Infection, Flow Cytometry, Cytometry