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Merck KGaA mononuclear cells mncs
Mononuclear Cells Mncs, 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/mononuclear cells mncs/product/Merck KGaA
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mononuclear cells mncs - by Bioz Stars, 2021-03
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Isolation:

Article Title: Simple Physical Model Unravels Influences of Chemokine on Shape Deformation and Migration of Human Hematopoietic Stem Cells
Article Snippet: .. Mononuclear cells (MNCs) were isolated by density-gradient centrifugation (Merck KGaA, Darmstadt, Germany), and CD34+ cells enriched by magnetic beads were further sorted (2×) by using an AutoMACS affinity column (all Miltenyi Biotec GmbH, Bergisch-Gladbach, Germany). .. Non-viable cells were removed by propidium iodide staining.

Article Title: Dynamic cellular phynotyping defines specific mobilization mechanisms of human hematopoietic stem and progenitor cells induced by SDF1α versus synthetic agents
Article Snippet: HSPC were isolated following the previous accounts , . .. In brief: mononuclear cells (MNCs) were isolated by density-gradient centrifugation (Merck KGaA, Darmstadt, Germany). .. CD34+ cells from the MNC fraction were enriched by using magnetic beads, followed by 2× sorting steps using an AutoMACS affinity column (all Miltenyi Biotec GmbH, Bergisch-Gladbach, Germany).

Article Title: Quantifying Adhesion Mechanisms and Dynamics of Human Hematopoietic Stem and Progenitor Cells
Article Snippet: HSC were isolated as previously described . .. Briefly, mononuclear cells (MNCs) were isolated by density gradient centrifugation using the Ficoll–Hypaque technique (Merck KGaA, Darmstadt, Germany). .. CD34+ cells from the MNC fraction were enriched by labelling with magnetic microbeads and sorted twice using an affinity column with the AutoMACS system (all Miltenyi Biotec GmbH, Bergisch-Gladbach, Germany).

Article Title: Candida albicans β-Glucan Differentiates Human Monocytes Into a Specific Subset of Macrophages
Article Snippet: .. 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).

Centrifugation:

Article Title: Simple Physical Model Unravels Influences of Chemokine on Shape Deformation and Migration of Human Hematopoietic Stem Cells
Article Snippet: .. Mononuclear cells (MNCs) were isolated by density-gradient centrifugation (Merck KGaA, Darmstadt, Germany), and CD34+ cells enriched by magnetic beads were further sorted (2×) by using an AutoMACS affinity column (all Miltenyi Biotec GmbH, Bergisch-Gladbach, Germany). .. Non-viable cells were removed by propidium iodide staining.

Article Title: Dynamic cellular phynotyping defines specific mobilization mechanisms of human hematopoietic stem and progenitor cells induced by SDF1α versus synthetic agents
Article Snippet: HSPC were isolated following the previous accounts , . .. In brief: mononuclear cells (MNCs) were isolated by density-gradient centrifugation (Merck KGaA, Darmstadt, Germany). .. CD34+ cells from the MNC fraction were enriched by using magnetic beads, followed by 2× sorting steps using an AutoMACS affinity column (all Miltenyi Biotec GmbH, Bergisch-Gladbach, Germany).

Magnetic Beads:

Article Title: Simple Physical Model Unravels Influences of Chemokine on Shape Deformation and Migration of Human Hematopoietic Stem Cells
Article Snippet: .. Mononuclear cells (MNCs) were isolated by density-gradient centrifugation (Merck KGaA, Darmstadt, Germany), and CD34+ cells enriched by magnetic beads were further sorted (2×) by using an AutoMACS affinity column (all Miltenyi Biotec GmbH, Bergisch-Gladbach, Germany). .. Non-viable cells were removed by propidium iodide staining.

Affinity Column:

Article Title: Simple Physical Model Unravels Influences of Chemokine on Shape Deformation and Migration of Human Hematopoietic Stem Cells
Article Snippet: .. Mononuclear cells (MNCs) were isolated by density-gradient centrifugation (Merck KGaA, Darmstadt, Germany), and CD34+ cells enriched by magnetic beads were further sorted (2×) by using an AutoMACS affinity column (all Miltenyi Biotec GmbH, Bergisch-Gladbach, Germany). .. Non-viable cells were removed by propidium iodide staining.

Small Interfering RNA:

Article Title: CSF1R inhibitors exhibit antitumor activity in acute myeloid leukemia by blocking paracrine signals from support cells
Article Snippet: Primary AML samples were obtained from patients by informed consent according to a protocol approved by the Oregon Health & Science University Institutional Review Board, and processed as described previously., The half-maximal inhibitory concentration (IC50) and area under the curve (AUC) were determined for each sample using probit regression analysis (see supplemental Methods, available on the Blood Web site). .. Within each patient sample, a small-interfering RNA (siRNA) “hit” was identified if its cell viability was at least 2 standard deviations less than the mean computed across all siRNAs tested (z score ≤ −2)., To evaluate apoptosis, mononuclear cells (MNCs) were exposed to either GW-2580 or ARRY-382 at 10 µM, and apoptosis was measured after 24, 48, and 72 hours by Annexin V staining (Guava Nexin assay; Merck Millipore, Billerica, MA). ..

Staining:

Article Title: CSF1R inhibitors exhibit antitumor activity in acute myeloid leukemia by blocking paracrine signals from support cells
Article Snippet: Primary AML samples were obtained from patients by informed consent according to a protocol approved by the Oregon Health & Science University Institutional Review Board, and processed as described previously., The half-maximal inhibitory concentration (IC50) and area under the curve (AUC) were determined for each sample using probit regression analysis (see supplemental Methods, available on the Blood Web site). .. Within each patient sample, a small-interfering RNA (siRNA) “hit” was identified if its cell viability was at least 2 standard deviations less than the mean computed across all siRNAs tested (z score ≤ −2)., To evaluate apoptosis, mononuclear cells (MNCs) were exposed to either GW-2580 or ARRY-382 at 10 µM, and apoptosis was measured after 24, 48, and 72 hours by Annexin V staining (Guava Nexin assay; Merck Millipore, Billerica, MA). ..

Gradient Centrifugation:

Article Title: Quantifying Adhesion Mechanisms and Dynamics of Human Hematopoietic Stem and Progenitor Cells
Article Snippet: HSC were isolated as previously described . .. Briefly, mononuclear cells (MNCs) were isolated by density gradient centrifugation using the Ficoll–Hypaque technique (Merck KGaA, Darmstadt, Germany). .. CD34+ cells from the MNC fraction were enriched by labelling with magnetic microbeads and sorted twice using an affinity column with the AutoMACS system (all Miltenyi Biotec GmbH, Bergisch-Gladbach, Germany).

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). ..

Article Title: Candida albicans β-Glucan Differentiates Human Monocytes Into a Specific Subset of Macrophages
Article Snippet: .. 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).

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
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    86
    Merck KGaA ifn γ staining effector peripheral blood mononuclear cells
    <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.
    Ifn γ Staining Effector Peripheral Blood Mononuclear Cells, 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/ifn γ staining effector peripheral blood mononuclear cells/product/Merck KGaA
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
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    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 mononuclear cells mncs
    <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.
    Mononuclear Cells Mncs, 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/mononuclear cells mncs/product/Merck KGaA
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mononuclear cells mncs - by Bioz Stars, 2021-03
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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