Effect of kinase inhibitors on nitric oxide production in Salmonella-infected chicken macrophage HD-11 cells.

<p>The HD-11 cells were infected with S. Enteritidis as described in the Materials and Methods and followed by treatment with various inhibitors at concentrations indicated. Nitrite in the culture media was measured at 24 hpi. Data are means ± standard deviations. The symbol (*) indicates the differences between the control and the treatments are statistically significant (P≤0.05).</p

Effect of H-89 on <i>S</i>. Enteritidis growth in trypticase soy broth (TSB).

<p>A. The growth curves of <i>S</i>. Enteritidis in TSB containing various concentrations of H-89. An overnight culture of <i>S</i>. Enteritidis was diluted 1∶20 in TSB medium containing various concentrations of H-89 and incubated at 41°C in a 96-well optical plate. The optical densities at OD600 were measured at 2, 4, 8, and 24 h using a microplate reader to determine the growth of <i>S</i>. Enteritidis. B. The viable <i>S</i>. Enteritidis in culture containing various concentrations of H-89. At the beginning (2 hpi) and the end of incubation (24 hpi), viable <i>S</i>. Enteritidis [Log10 (cfu)] in each treatment were determined by plating serial 1∶10 dilutions of the culture on the Difco’s xylose-lysine tergitol 4 (XLT4) agar plates and incubated at 41°C for 24 h.</p

Effect of H-89 treatment on nitric oxide (NO) production in <i>S</i>. Enteritidis-infected HD11 cells and the intracellular survival of <i>S</i>. Enteritidis.

<p>A. Effect of H-89 treatment on NO production in <i>S</i>. Enteritidis-infected HD11 cells. HD11 cells were infected with live <i>S</i>. Enteritidis (SE) for 1 h in 24-well plates at 39°C in a 5% CO2 humidified incubator. At 1 hour post infection (hpi), extracellular SE were killed by incubation with media containing 100 µg/mL of gentamicin sulfate for 1 h; the cells were washed and then cultured in media containing various concentrations of H-89 and 20 µg/mL of gentamicin sulfate for an additional 22 h; and nitrite contents in cell culture media were determined. Treatment with heat-killed <i>S</i>. Enteritidis (HKSE) was performed identically as with live SE. B. HD11 cells were infected with live SE as described above and at 24 hpi, intracellular viable SE [Log10 (cfu)] were counted. The symbol (*) indicates that the difference between these groups and the respective controls is statistically significant (p<0.05).</p

Intracellular <i>S</i>. Enteritidis in HD11 cells infected with various multiplicity of infection (MOI).

<p>HD11 cells were infected with <i>S</i>. Enteritidis at various MOI for 1 h in 24-well plates at 39°C in a 5% CO2 humidified incubator. At 1 hour post infection (hpi), extracellular <i>S</i>. Enteritidis were killed by incubation with media containing 100 µg/mL of gentamicin sulfate for 1 h. Intracellular <i>S</i>. Enteritidis (cfu) were determined at 2 and 24 hpi. A. Intracellular <i>S</i>. Enteritidis (cfu) at 2 hpi; B. Intracellular <i>S</i>. Enteritidis (cfu) at 24 hpi.</p

Effect of <i>S</i>. Enteritidis infection on nitric oxide (NO) production in HD11 cells.

<p>HD11 cells were infected with live <i>S.</i> Enteritidis (SE) for 1 h in 24-well plates at 39°C in a 5% CO2 humidified incubator. At 1 hour post infection (hpi), extracellular SE were killed by incubation with media containing 100 µg/mL of gentamicin sulfate for 1 h; the cells were washed and then cultured with or without lipopolysaccharide (LPS) at 0.2 µg/mL for an additional 22 h in a medium containing 20 µg/mL of gentamicin sulfate; and nitrite contents in cell culture media were determined. Treatment with heat-killed <i>S</i>. Enteritidis (HKSE) was performed identically as with live SE. The symbol (*) indicates that the difference between these groups and the control is statistically significant (p<0.05).</p

Boxplots of log₁₀ values of mRNA copy numbers by age and treatment group: blue boxes are data from the control (saline) group and red boxes are data from the CpG-treated foals.

<p>The triangles in the middle of boxes represent the median value; the bottom and top of the boxes represent the 25^th and 75^th percentiles, respectively. The vertical lines extending from the boxes to horizontal lines represent multiples of 1.75 of the respective interquartile distance. Numbers above boxes represent within-treatment group differences among ages: within treatment group, ages with different numbers differed significantly (P<0.05) between groups. Asterisks denote days on which values were significantly (P<0.05) different between the control and Cp-G-treated foals. Arrows along the horizontal axis indicate ages when treatment (saline or CpG) was administered.</p

Mean (and 95% confidence intervals) for neutrophil parameters determined using flow cytometry (please see text for details) and estimated by mixed-effects modeling.

<p>Values in columns with the same letter indicate absence of statistical significance between groups for a given age. Values in rows with differing superscripted numbers indicate significant (P<0.05) differences among ages within group.</p><p>Mean (and 95% confidence intervals) for neutrophil parameters determined using flow cytometry (please see text for details) and estimated by mixed-effects modeling.</p

Boxplots of values of liberated 4-methylumbelliferone (4-MU) measured fluorometrically by age and treatment group: blue boxes are data from the control (saline) group and red boxes are data from the CpG-treated foal (see <b>Figure 1</b> for boxplot description).

<p>Numbers above boxes represent within-treatment group differences among ages: within treatment group, ages with different numbers differed significantly (P<0.05) between groups. Asterisks denote days on which values were significantly (P<0.05) different between the control and Cp-G-treated foals. Arrows along the horizontal axis indicate ages when treatment (saline or CpG was administered.</p

Mean (and 95% confidence intervals) for mRNA copy numbers estimated by mixed-effects modeling for IL-4, IL-6, and IL-8.

<p>Data represent back-transformed results of log₁₀-transformed data. Values in columns with the same letter indicate absence of statistical significance between groups for a given age. Values in rows with differing superscripted numbers indicate significant (P<0.05) differences among ages within group.</p><p>Mean (and 95% confidence intervals) for mRNA copy numbers estimated by mixed-effects modeling for IL-4, IL-6, and IL-8.</p