human peripheral blood mononuclear cells mnc  (Merck KGaA)

 
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    Merck KGaA human peripheral blood mononuclear cells mnc
    Human Peripheral Blood Mononuclear Cells Mnc, supplied by Merck KGaA, 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/human peripheral blood mononuclear cells mnc/product/Merck KGaA
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    human peripheral blood mononuclear cells mnc - by Bioz Stars, 2021-03
    86/100 stars

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    Isolation:

    Article Title: Salt-responsive gut commensal modulates TH17 axis and disease
    Article Snippet: .. Peripheral blood mononuclear cells (PBMCs) were immediately isolated by density centrifugation using Biocoll (Merck, Darmstadt, Germany). .. 106 CD4+ enriched cell fractions isolated by CD4+ T Cell Kit (Miltenyi) were plated onto U-bottom plates and were restimulated for 4 hours at 37°C, 5% CO2 in a humidified incubator.

    Article Title: Candida albicans β-Glucan Differentiates Human Monocytes Into a Specific Subset of Macrophages
    Article Snippet: Isolation and culture of human monocytes Peripheral blood was collected from healthy, male, non-smoking volunteers after obtaining informed consent and approval by the Institutional Ethics Committee. .. Blood mononuclear cells (PBMCs) were isolated using density gradient centrifugation (Biocoll, Merck Millipore). .. Classical monocytes (CD14++ CD16− ) were purified by negative selection (Dynabeads Untouched Human Monocytes Kit, Thermo Fisher Scientific).

    Article Title: Apolipoprotein A-I gene transfer exerts immunomodulatory effects and reduces vascular inflammation and fibrosis in ob/ob mice
    Article Snippet: Human peripheral blood mononuclear cell isolation Blood was withdrawn from healthy donors (aged 40 to 65) with approval (EA4/115/11) from the Ethical Commission (Charité, CBF, Berlin). .. Human peripheral blood mononuclear cells (MNC) were isolated from the blood samples by density-gradient centrifugation (Biocoll; Merck Millipore, Darmstadt, Germany). .. Adhesion assay HAEC were plated in gelatin/fibronectin-coated black solid bottom 96-well plates, cultured for 24 h and treated with 10 ng/ml TNF-α (BD Pharmingen, Franklin Lakes, NJ, USA) in Endothelial Basal Medium for 4 h prior to addition of MNC.

    Article Title: CARD8 inflammasome activation triggers pyroptosis in human T cells
    Article Snippet: While these observations do not yet rule out that GSDMD triggered cell death in T cells is pro‐inflammatory and that T cells can acquire expression of IL‐1‐related cytokines under certain pathogenic circumstances, these current data would indeed imply a complete disconnect of “pyroptosis” from its initial definition. .. Isolation of PBMCs, primary human monocytes and T cellsPeripheral blood mononuclear cells (PBMCs) were isolated from residual heparinized blood retained in leukocyte reduction system chambers of healthy thrombocyte donors by density gradient centrifugation (Biocoll, Merck Millipore) and erythrocyte lysis (RBC lysis buffer, BioLegend). ..

    Centrifugation:

    Article Title: Salt-responsive gut commensal modulates TH17 axis and disease
    Article Snippet: .. Peripheral blood mononuclear cells (PBMCs) were immediately isolated by density centrifugation using Biocoll (Merck, Darmstadt, Germany). .. 106 CD4+ enriched cell fractions isolated by CD4+ T Cell Kit (Miltenyi) were plated onto U-bottom plates and were restimulated for 4 hours at 37°C, 5% CO2 in a humidified incubator.

    Article Title: Apolipoprotein A-I gene transfer exerts immunomodulatory effects and reduces vascular inflammation and fibrosis in ob/ob mice
    Article Snippet: Human peripheral blood mononuclear cell isolation Blood was withdrawn from healthy donors (aged 40 to 65) with approval (EA4/115/11) from the Ethical Commission (Charité, CBF, Berlin). .. Human peripheral blood mononuclear cells (MNC) were isolated from the blood samples by density-gradient centrifugation (Biocoll; Merck Millipore, Darmstadt, Germany). .. Adhesion assay HAEC were plated in gelatin/fibronectin-coated black solid bottom 96-well plates, cultured for 24 h and treated with 10 ng/ml TNF-α (BD Pharmingen, Franklin Lakes, NJ, USA) in Endothelial Basal Medium for 4 h prior to addition of MNC.

    Gradient Centrifugation:

    Article Title: Candida albicans β-Glucan Differentiates Human Monocytes Into a Specific Subset of Macrophages
    Article Snippet: Isolation and culture of human monocytes Peripheral blood was collected from healthy, male, non-smoking volunteers after obtaining informed consent and approval by the Institutional Ethics Committee. .. Blood mononuclear cells (PBMCs) were isolated using density gradient centrifugation (Biocoll, Merck Millipore). .. Classical monocytes (CD14++ CD16− ) were purified by negative selection (Dynabeads Untouched Human Monocytes Kit, Thermo Fisher Scientific).

    Article Title: CARD8 inflammasome activation triggers pyroptosis in human T cells
    Article Snippet: While these observations do not yet rule out that GSDMD triggered cell death in T cells is pro‐inflammatory and that T cells can acquire expression of IL‐1‐related cytokines under certain pathogenic circumstances, these current data would indeed imply a complete disconnect of “pyroptosis” from its initial definition. .. Isolation of PBMCs, primary human monocytes and T cellsPeripheral blood mononuclear cells (PBMCs) were isolated from residual heparinized blood retained in leukocyte reduction system chambers of healthy thrombocyte donors by density gradient centrifugation (Biocoll, Merck Millipore) and erythrocyte lysis (RBC lysis buffer, BioLegend). ..

    Article Title: The Lipid A from the Lipopolysaccharide of the Phototrophic Bacterium Rhodomicrobium vannielii ATCC 17100 Revisited
    Article Snippet: .. Assay in Human Mononuclear Cells (MNC) Peripheral blood MNC from healthy human volunteers (prepared from heparinized blood by gradient centrifugation while using Biocoll, Merck, Darmstadt, Germany) were incubated at a concentration of 1 × 106 /mL in 96-well tissue culture plates at a volume of 150 μL using RPMI-1640 medium that was supplemented with 100 U/mL penicillin, 100 μg/mL streptomycin (both PAA Laboratories, GmbH, Cölbe, Germany), and 10% of heat-inactivated FCS (Merck Millipore, Biochrom AG, Berlin, Germany). ..

    Lysis:

    Article Title: CARD8 inflammasome activation triggers pyroptosis in human T cells
    Article Snippet: While these observations do not yet rule out that GSDMD triggered cell death in T cells is pro‐inflammatory and that T cells can acquire expression of IL‐1‐related cytokines under certain pathogenic circumstances, these current data would indeed imply a complete disconnect of “pyroptosis” from its initial definition. .. Isolation of PBMCs, primary human monocytes and T cellsPeripheral blood mononuclear cells (PBMCs) were isolated from residual heparinized blood retained in leukocyte reduction system chambers of healthy thrombocyte donors by density gradient centrifugation (Biocoll, Merck Millipore) and erythrocyte lysis (RBC lysis buffer, BioLegend). ..

    Incubation:

    Article Title: The Lipid A from the Lipopolysaccharide of the Phototrophic Bacterium Rhodomicrobium vannielii ATCC 17100 Revisited
    Article Snippet: .. Assay in Human Mononuclear Cells (MNC) Peripheral blood MNC from healthy human volunteers (prepared from heparinized blood by gradient centrifugation while using Biocoll, Merck, Darmstadt, Germany) were incubated at a concentration of 1 × 106 /mL in 96-well tissue culture plates at a volume of 150 μL using RPMI-1640 medium that was supplemented with 100 U/mL penicillin, 100 μg/mL streptomycin (both PAA Laboratories, GmbH, Cölbe, Germany), and 10% of heat-inactivated FCS (Merck Millipore, Biochrom AG, Berlin, Germany). ..

    Concentration Assay:

    Article Title: The Lipid A from the Lipopolysaccharide of the Phototrophic Bacterium Rhodomicrobium vannielii ATCC 17100 Revisited
    Article Snippet: .. Assay in Human Mononuclear Cells (MNC) Peripheral blood MNC from healthy human volunteers (prepared from heparinized blood by gradient centrifugation while using Biocoll, Merck, Darmstadt, Germany) were incubated at a concentration of 1 × 106 /mL in 96-well tissue culture plates at a volume of 150 μL using RPMI-1640 medium that was supplemented with 100 U/mL penicillin, 100 μg/mL streptomycin (both PAA Laboratories, GmbH, Cölbe, Germany), and 10% of heat-inactivated FCS (Merck Millipore, Biochrom AG, Berlin, Germany). ..

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    Merck KGaA human pbmc derived macrophages
    <t>CLEC5A</t> mediates enhanced proinflammatory cytokine and chemokine induction in human M-Mϕ after influenza virus infection. M-Mϕ differentiated from the <t>PBMC</t> of 8 independent donors were transfected with CLEC5A gene-specific or nontargeting siRNA, followed by cell sorting to obtain the CLEC5A-positive (CLEC5A_pos) and CLEC5A-negative (CLEC5A_neg) populations. The macrophages were then infected with HK HA,NA or VN HA,NA recombinant viruses at an MOI of 2 for 24 h to determine viral M gene copy numbers in infected M-Mϕ and viral titers in culture supernatants (log 10 TCID 50 /ml in MDCK cells) (A) and proinflammatory cytokines and chemokines in culture supernatant (means ± SD, in pg/ml) (B). P values from Mann-Whitney tests are shown.
    Human Pbmc Derived Macrophages, supplied by Merck KGaA, 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/human pbmc derived macrophages/product/Merck KGaA
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    human pbmc derived macrophages - by Bioz Stars, 2021-03
    86/100 stars
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    86
    Merck KGaA blood mononuclear cells pbmcs
    <t>CLEC5A</t> mediates enhanced proinflammatory cytokine and chemokine induction in human M-Mϕ after influenza virus infection. M-Mϕ differentiated from the <t>PBMC</t> of 8 independent donors were transfected with CLEC5A gene-specific or nontargeting siRNA, followed by cell sorting to obtain the CLEC5A-positive (CLEC5A_pos) and CLEC5A-negative (CLEC5A_neg) populations. The macrophages were then infected with HK HA,NA or VN HA,NA recombinant viruses at an MOI of 2 for 24 h to determine viral M gene copy numbers in infected M-Mϕ and viral titers in culture supernatants (log 10 TCID 50 /ml in MDCK cells) (A) and proinflammatory cytokines and chemokines in culture supernatant (means ± SD, in pg/ml) (B). P values from Mann-Whitney tests are shown.
    Blood Mononuclear Cells Pbmcs, supplied by Merck KGaA, 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/blood mononuclear cells pbmcs/product/Merck KGaA
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    blood mononuclear cells pbmcs - by Bioz Stars, 2021-03
    86/100 stars
      Buy from Supplier

    86
    Merck KGaA peripheral blood mononuclear cells pbmcs
    <t>CLEC5A</t> mediates enhanced proinflammatory cytokine and chemokine induction in human M-Mϕ after influenza virus infection. M-Mϕ differentiated from the <t>PBMC</t> of 8 independent donors were transfected with CLEC5A gene-specific or nontargeting siRNA, followed by cell sorting to obtain the CLEC5A-positive (CLEC5A_pos) and CLEC5A-negative (CLEC5A_neg) populations. The macrophages were then infected with HK HA,NA or VN HA,NA recombinant viruses at an MOI of 2 for 24 h to determine viral M gene copy numbers in infected M-Mϕ and viral titers in culture supernatants (log 10 TCID 50 /ml in MDCK cells) (A) and proinflammatory cytokines and chemokines in culture supernatant (means ± SD, in pg/ml) (B). P values from Mann-Whitney tests are shown.
    Peripheral Blood Mononuclear Cells Pbmcs, supplied by Merck KGaA, 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/peripheral blood mononuclear cells pbmcs/product/Merck KGaA
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    peripheral blood mononuclear cells pbmcs - by Bioz Stars, 2021-03
    86/100 stars
      Buy from Supplier

    86
    Merck KGaA human peripheral blood mononuclear cells mnc
    <t>CLEC5A</t> mediates enhanced proinflammatory cytokine and chemokine induction in human M-Mϕ after influenza virus infection. M-Mϕ differentiated from the <t>PBMC</t> of 8 independent donors were transfected with CLEC5A gene-specific or nontargeting siRNA, followed by cell sorting to obtain the CLEC5A-positive (CLEC5A_pos) and CLEC5A-negative (CLEC5A_neg) populations. The macrophages were then infected with HK HA,NA or VN HA,NA recombinant viruses at an MOI of 2 for 24 h to determine viral M gene copy numbers in infected M-Mϕ and viral titers in culture supernatants (log 10 TCID 50 /ml in MDCK cells) (A) and proinflammatory cytokines and chemokines in culture supernatant (means ± SD, in pg/ml) (B). P values from Mann-Whitney tests are shown.
    Human Peripheral Blood Mononuclear Cells Mnc, supplied by Merck KGaA, 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/human peripheral blood mononuclear cells mnc/product/Merck KGaA
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    human peripheral blood mononuclear cells mnc - by Bioz Stars, 2021-03
    86/100 stars
      Buy from Supplier

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    CLEC5A mediates enhanced proinflammatory cytokine and chemokine induction in human M-Mϕ after influenza virus infection. M-Mϕ differentiated from the PBMC of 8 independent donors were transfected with CLEC5A gene-specific or nontargeting siRNA, followed by cell sorting to obtain the CLEC5A-positive (CLEC5A_pos) and CLEC5A-negative (CLEC5A_neg) populations. The macrophages were then infected with HK HA,NA or VN HA,NA recombinant viruses at an MOI of 2 for 24 h to determine viral M gene copy numbers in infected M-Mϕ and viral titers in culture supernatants (log 10 TCID 50 /ml in MDCK cells) (A) and proinflammatory cytokines and chemokines in culture supernatant (means ± SD, in pg/ml) (B). P values from Mann-Whitney tests are shown.

    Journal: Journal of Virology

    Article Title: CLEC5A-Mediated Enhancement of the Inflammatory Response in Myeloid Cells Contributes to Influenza Virus Pathogenicity In Vivo

    doi: 10.1128/JVI.01813-16

    Figure Lengend Snippet: CLEC5A mediates enhanced proinflammatory cytokine and chemokine induction in human M-Mϕ after influenza virus infection. M-Mϕ differentiated from the PBMC of 8 independent donors were transfected with CLEC5A gene-specific or nontargeting siRNA, followed by cell sorting to obtain the CLEC5A-positive (CLEC5A_pos) and CLEC5A-negative (CLEC5A_neg) populations. The macrophages were then infected with HK HA,NA or VN HA,NA recombinant viruses at an MOI of 2 for 24 h to determine viral M gene copy numbers in infected M-Mϕ and viral titers in culture supernatants (log 10 TCID 50 /ml in MDCK cells) (A) and proinflammatory cytokines and chemokines in culture supernatant (means ± SD, in pg/ml) (B). P values from Mann-Whitney tests are shown.

    Article Snippet: After 1 h of adsorption, the virus inoculum was removed and the cells were washed once with phosphate-buffered saline (PBS), followed by incubation with infection medium (RPMI supplemented with 2% FBS) for 24 h. For Syk inhibition and CLEC5A blocking assay, human PBMC-derived macrophages were pretreated with the indicated concentrations of Bay 61-3606 (Merck Millipore) or anti-human CLEC5A antagonistic antibody for 1 h before and throughout the infection process.

    Techniques: Infection, Transfection, FACS, Recombinant, MANN-WHITNEY

    CLEC5A mediates enhanced proinflammatory cytokine and chemokine induction in human GM-Mϕ after influenza virus infection. GM-Mϕ differentiated from the PBMC of 2 independent donors were transfected with CLEC5A gene-specific or nontargeting siRNA, followed by cell sorting to obtain the CLEC5A-positive (CLEC5A_pos) and CLEC5A-negative (CLEC5A_neg) populations. The macrophages were then infected with HK HA,NA or VN HA,NA recombinant viruses at an MOI of 2 for 24 h to determine viral M gene copy numbers in infected GM-Mϕ and viral titers in culture supernatant (log 10 TCID 50 /ml in MDCK cells) (A) and proinflammatory cytokines and chemokines in culture supernatants (means ± SD, in pg/ml) (B). P values from Mann-Whitney tests are shown.

    Journal: Journal of Virology

    Article Title: CLEC5A-Mediated Enhancement of the Inflammatory Response in Myeloid Cells Contributes to Influenza Virus Pathogenicity In Vivo

    doi: 10.1128/JVI.01813-16

    Figure Lengend Snippet: CLEC5A mediates enhanced proinflammatory cytokine and chemokine induction in human GM-Mϕ after influenza virus infection. GM-Mϕ differentiated from the PBMC of 2 independent donors were transfected with CLEC5A gene-specific or nontargeting siRNA, followed by cell sorting to obtain the CLEC5A-positive (CLEC5A_pos) and CLEC5A-negative (CLEC5A_neg) populations. The macrophages were then infected with HK HA,NA or VN HA,NA recombinant viruses at an MOI of 2 for 24 h to determine viral M gene copy numbers in infected GM-Mϕ and viral titers in culture supernatant (log 10 TCID 50 /ml in MDCK cells) (A) and proinflammatory cytokines and chemokines in culture supernatants (means ± SD, in pg/ml) (B). P values from Mann-Whitney tests are shown.

    Article Snippet: After 1 h of adsorption, the virus inoculum was removed and the cells were washed once with phosphate-buffered saline (PBS), followed by incubation with infection medium (RPMI supplemented with 2% FBS) for 24 h. For Syk inhibition and CLEC5A blocking assay, human PBMC-derived macrophages were pretreated with the indicated concentrations of Bay 61-3606 (Merck Millipore) or anti-human CLEC5A antagonistic antibody for 1 h before and throughout the infection process.

    Techniques: Infection, Transfection, FACS, Recombinant, MANN-WHITNEY

    CLEC5A-mediated proinflammatory response in human M-Mϕ after infections with influenza viruses of H1N1, H5N1, and H7N9 subtypes. Human M-Mϕ differentiated from PBMC of 2 independent donors were infected with A/Hong Kong/54/98 (H1N1), A/Vietnam/1203/04 (H5N1), or A/Shanghai/2/13 (H7N9) influenza viruses at an MOI of 2 for 24 h to determine viral M gene copy numbers in infected M-Mϕ (A) and proinflammatory cytokines and chemokines in culture supernatants (means ± SD, in pg/ml) (B). P values from Mann-Whitney tests are shown.

    Journal: Journal of Virology

    Article Title: CLEC5A-Mediated Enhancement of the Inflammatory Response in Myeloid Cells Contributes to Influenza Virus Pathogenicity In Vivo

    doi: 10.1128/JVI.01813-16

    Figure Lengend Snippet: CLEC5A-mediated proinflammatory response in human M-Mϕ after infections with influenza viruses of H1N1, H5N1, and H7N9 subtypes. Human M-Mϕ differentiated from PBMC of 2 independent donors were infected with A/Hong Kong/54/98 (H1N1), A/Vietnam/1203/04 (H5N1), or A/Shanghai/2/13 (H7N9) influenza viruses at an MOI of 2 for 24 h to determine viral M gene copy numbers in infected M-Mϕ (A) and proinflammatory cytokines and chemokines in culture supernatants (means ± SD, in pg/ml) (B). P values from Mann-Whitney tests are shown.

    Article Snippet: After 1 h of adsorption, the virus inoculum was removed and the cells were washed once with phosphate-buffered saline (PBS), followed by incubation with infection medium (RPMI supplemented with 2% FBS) for 24 h. For Syk inhibition and CLEC5A blocking assay, human PBMC-derived macrophages were pretreated with the indicated concentrations of Bay 61-3606 (Merck Millipore) or anti-human CLEC5A antagonistic antibody for 1 h before and throughout the infection process.

    Techniques: Infection, MANN-WHITNEY

    Blocking CLEC5A-mediated signaling with Syk inhibitor (Bay 61-3606) and CLEC5A antagonizing antibodies reduced inflammatory response in human M-Mϕ after influenza virus infection. (A) Human M-Mϕ differentiated from the PBMC of 3 independent donors were treated with dimethyl sulfoxide or the Syk inhibitor Bay 61-3606 prior to and throughout the course of infection with HK HA,NA or VN HA,NA virus at an MOI of 2 to determine the TNF-α and IP-10 concentrations (means ± SD, in pg/ml) from culture supernatants at 24 h postinfection. (B) Human M-Mϕ differentiated from the PBMC of 4 independent donors were incubated with 1 μg/ml of murine IgG1 isotype control antibody (Iso ctrl) or clones of anti-human CLEC5A antibodies, which were previously shown to possess antagonizing activity against DV or JEV infections, prior to and throughout the course of infection with VN HA,NA virus at an MOI of 2 to determine the level of IP-10 (means ± SD, in pg/ml) from culture supernatants at 24 h postinfection. (C) Human M-Mϕ differentiated from the PBMC of 4 independent donors were incubated with increasing concentrations of murine IgG1 isotype control antibody or the anti-CLEC5A antibody (clone 8H8F5) prior to and throughout the course of infection with VN HA,NA virus at an MOI of 2 to determine the level of IP-10 (means ± SD, in pg/ml) from culture supernatants at 24 h postinfection. ns, not significant. (D) M-Mϕ differentiated from the PBMC of 2 independent donors were transfected with CLEC5A gene-specific or nontargeting siRNA followed by cell sorting to obtain the CLEC5A-positive (CLEC5A_pos) and CLEC5A-negative (CLEC5A_neg) populations. M-Mϕ were infected with VN HA,NA or UV-inactivated VN HA,NA (UV-VN HA,NA ) at an MOI of 2. Viral M gene copy numbers and levels of TNF-α and IP-10 (means ± SD, in pg/ml) in culture supernatant were determined at 24 h postinfection. P values from Mann-Whitney tests are shown.

    Journal: Journal of Virology

    Article Title: CLEC5A-Mediated Enhancement of the Inflammatory Response in Myeloid Cells Contributes to Influenza Virus Pathogenicity In Vivo

    doi: 10.1128/JVI.01813-16

    Figure Lengend Snippet: Blocking CLEC5A-mediated signaling with Syk inhibitor (Bay 61-3606) and CLEC5A antagonizing antibodies reduced inflammatory response in human M-Mϕ after influenza virus infection. (A) Human M-Mϕ differentiated from the PBMC of 3 independent donors were treated with dimethyl sulfoxide or the Syk inhibitor Bay 61-3606 prior to and throughout the course of infection with HK HA,NA or VN HA,NA virus at an MOI of 2 to determine the TNF-α and IP-10 concentrations (means ± SD, in pg/ml) from culture supernatants at 24 h postinfection. (B) Human M-Mϕ differentiated from the PBMC of 4 independent donors were incubated with 1 μg/ml of murine IgG1 isotype control antibody (Iso ctrl) or clones of anti-human CLEC5A antibodies, which were previously shown to possess antagonizing activity against DV or JEV infections, prior to and throughout the course of infection with VN HA,NA virus at an MOI of 2 to determine the level of IP-10 (means ± SD, in pg/ml) from culture supernatants at 24 h postinfection. (C) Human M-Mϕ differentiated from the PBMC of 4 independent donors were incubated with increasing concentrations of murine IgG1 isotype control antibody or the anti-CLEC5A antibody (clone 8H8F5) prior to and throughout the course of infection with VN HA,NA virus at an MOI of 2 to determine the level of IP-10 (means ± SD, in pg/ml) from culture supernatants at 24 h postinfection. ns, not significant. (D) M-Mϕ differentiated from the PBMC of 2 independent donors were transfected with CLEC5A gene-specific or nontargeting siRNA followed by cell sorting to obtain the CLEC5A-positive (CLEC5A_pos) and CLEC5A-negative (CLEC5A_neg) populations. M-Mϕ were infected with VN HA,NA or UV-inactivated VN HA,NA (UV-VN HA,NA ) at an MOI of 2. Viral M gene copy numbers and levels of TNF-α and IP-10 (means ± SD, in pg/ml) in culture supernatant were determined at 24 h postinfection. P values from Mann-Whitney tests are shown.

    Article Snippet: After 1 h of adsorption, the virus inoculum was removed and the cells were washed once with phosphate-buffered saline (PBS), followed by incubation with infection medium (RPMI supplemented with 2% FBS) for 24 h. For Syk inhibition and CLEC5A blocking assay, human PBMC-derived macrophages were pretreated with the indicated concentrations of Bay 61-3606 (Merck Millipore) or anti-human CLEC5A antagonistic antibody for 1 h before and throughout the infection process.

    Techniques: Blocking Assay, Infection, Incubation, Clone Assay, Activity Assay, Transfection, FACS, MANN-WHITNEY