Journal: BMC Microbiology

Article Title: Cluster analysis of host cytokine responses to biodefense pathogens in a whole blood ex vivo exposure model (WEEM)

doi: 10.1186/1471-2180-12-79

Figure Lengend Snippet: Scatter plots of 16 cytokine concentrations detected in human blood following ex vivo bacterial exposures. Cytokine concentrations were displayed on a logarithmic scale. The cytokines shown here were detected out of the 30 cytokines in the arrays. The 8 cytokines that were found to be statistically differentially expressed among these samples are highlighted with rectangular boxes. Each mark delineates the average of triplicate exposure samples. Each exposure sample is loaded onto a protein array chip that contains 5 independent measurements per cytokine meaning that fifteen measurements are used to obtain these data.

Article Snippet: These protein arrays allow the simultaneous quantification of 30 biologically relevant cytokines, as determined by Zyomyx, Inc: IL-1α, IL-1β, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IL-12(p40), IL-12(p40/p70), IL-12(p70), IL-13, IL-15, TNFα, TNFβ, Eotaxin, MCP-1, MCP-3, TRAIL, CD95(sFas), MIG, sICAM-1, IP-10, CD23, TGF-β, GM-CSF, GCSF, IFN-γ.

Techniques: Ex Vivo, Protein Array

Journal: BMC Microbiology

Article Title: Cluster analysis of host cytokine responses to biodefense pathogens in a whole blood ex vivo exposure model (WEEM)

doi: 10.1186/1471-2180-12-79

Figure Lengend Snippet: Concentrations of 8 cytokines in human whole blood after ex vivo exposure to pathogens. The control was a mock-exposed sample. Cytokine concentrations were determined using protein arrays. The bars represent the average of three replicate samples that each contain 5 replicate features per cytokine assay and the lines represent the standard deviation among the three replicates.

Article Snippet: These protein arrays allow the simultaneous quantification of 30 biologically relevant cytokines, as determined by Zyomyx, Inc: IL-1α, IL-1β, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IL-12(p40), IL-12(p40/p70), IL-12(p70), IL-13, IL-15, TNFα, TNFβ, Eotaxin, MCP-1, MCP-3, TRAIL, CD95(sFas), MIG, sICAM-1, IP-10, CD23, TGF-β, GM-CSF, GCSF, IFN-γ.

Techniques: Ex Vivo, Cytokine Assay, Standard Deviation

Journal: BMC Microbiology

Article Title: Cluster analysis of host cytokine responses to biodefense pathogens in a whole blood ex vivo exposure model (WEEM)

doi: 10.1186/1471-2180-12-79

Figure Lengend Snippet: Clustering result with HOPACH using the average linkage distance between clusters is shown. The eight pathogen-exposed samples are clustered according to the dendrogram on the left and cluster into five groups, 1) Y. pestis (KIM5, NYC, and India), 2) Y. pseudotuberculosis , 3) Y. enterocolitica , 4) B. anthracis (Ames and Sterne), and 5) Control. Sixteen cytokines (Eotaxin, IL-10, IL-12(p40), IL-15, IL-1α, IL-1β, IL-6, IL-8, IP-10, MCP-1, MIG, TNFα, TRAIL, sCD23, sCD95, and sICAM-1) are also reordered based on their correlations according to the dendrogram on the top. Clusters go from root at top to leaf node for each cytokine. Clusters in between are based on their agglomerative . The branch shows the similarity, the short the branch, the more similar. In addition, the eight rightmost proteins form a cluster that may involve inflammation-related cascades initiated by an innate immune response to these pathogen. Colors represent units of log10 [pg/ml], in ten equally spaced intervals increasing from white to dark red. A key showing the specific log10 values for each interval is shown in the figure.

Article Snippet: These protein arrays allow the simultaneous quantification of 30 biologically relevant cytokines, as determined by Zyomyx, Inc: IL-1α, IL-1β, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IL-12(p40), IL-12(p40/p70), IL-12(p70), IL-13, IL-15, TNFα, TNFβ, Eotaxin, MCP-1, MCP-3, TRAIL, CD95(sFas), MIG, sICAM-1, IP-10, CD23, TGF-β, GM-CSF, GCSF, IFN-γ.

Techniques:

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: Viral titers were determined in MDCK cells by determining the 50% tissue culture infective dose (TCID 50 ), and cytokines or chemokines were quantified by the bead-based cytokine quantification kit (BD Bioscience).

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

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: Viral titers were determined in MDCK cells by determining the 50% tissue culture infective dose (TCID 50 ), and cytokines or chemokines were quantified by the bead-based cytokine quantification kit (BD Bioscience).

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

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 −/− mice were protected from the sublethal challenge of VN HA,NA virus by reducing the level of proinflammatory cytokine in the lungs. WT and CLEC5A −/− mice were infected with 1 MLD 50 of VN HA,NA intranasally. Mouse survival (A) and body weight changes (B) were monitored every other day; P values from log-rank tests are shown. (C and D) Viral titer (means ± SD, in log 10 TCID 50 /ml) (C) and cytokine and chemokine levels (means ± SD, in pg/ml) (D) detected in the mouse lungs at day 1, 4, and 7 postinfection ( n = 4 per group at each time point); P values from Mann-Whitney tests are shown.

Article Snippet: Viral titers were determined in MDCK cells by determining the 50% tissue culture infective dose (TCID 50 ), and cytokines or chemokines were quantified by the bead-based cytokine quantification kit (BD Bioscience).

Techniques: Infection, MANN-WHITNEY