Light microscopy analysis of the effect secretions on the differentiation of monocytes to macrophages.

<p>Monocytes were differentiated to MØ-1 in the presence of GM-CSF (A) or in the presence of GM-CSF and 35 µg of secretions/ml (B) and their morphology evaluated. Similarly, monocytes were differentiated to MØ-2 in the presence of M-CSF (C) and in the presence of M-CSF and secretions (D). Results indicated in days are from a representative experiment.</p

LPS-induced growth factor production by secretions differentiated macrophages.

<p>The results for control macrophages are expressed as the median value and range, and are set at 100%. The effect of secretions is expressed as a percentage relative to the production of growth factors by control cells. Results are means±SEM of at least six experiments. Values are significantly (*p<0.05 and **p<0.005) different from those for macrophages stimulated with LPS. ND: not detectable.</p

LPS-induced cytokine production by secretions differentiated macrophages.

<p>The results for the control macrophages are expressed as the median value and range, and are set at 100%. The effect of secretions is expressed as a percentage relative to the cytokine production by control cells. Results are means±SEM of at least six experiments. Values are significantly (*p<0.05 and **p<0.005) different from those for macrophages stimulated with LPS.</p

Cytokine production in response to LTA.

<p>We measured the production of IL-12p40 (A,B), TNF-α (C,D), IL-6 (E,F) and IL-10 (G,H) by control and secretions-differentiated MØ-1 and MØ-2 induced by a range of LTA. The results, expressed in ng/ml, are means±SEM of 12 experiments. Open bars: control macrophages; filled bars: secretions differentiated macrophages. *p<0.05 for differences from control macrophages.</p

Expression of surface receptors by secretions-differentiated macrophages.

<p>Results, expressed as the mean fluorescence intensity (MFI), are means±SEM of 6–11 experiments. Values are significantly (*p<0.05 and **p<0.005) different from those for control cells. ND: not detectable.</p

LPS-induced chemokine production by secretions differentiated macrophages.

<p>The results for control macrophages are expressed as the median value and range, and are set at 100%. The effect of secretions is expressed as a percentage relative to the chemokine production by control cells. Results are means±SEM of at least six experiments. Values are significantly (*p<0.05 and **p<0.005) different from those for macrophages stimulated with LPS. ND: not detectable.</p

Cytokine production in response to LPS.

<p>We measured the production of IL-12p40 (A,B) and IL-10 (C,D) by control and secretions-differentiated MØ-1 and MØ-2 in response to a range of LPS. The results, expressed in ng/ml, are means±SEM of 9–10 experiments. Open bars: control macrophages; filled bars: secretions differentiated macrophages. *p<0.05 for differences from control macrophages.</p

IFN-γ in supernatants of IL-23 plus IL-1β stimulated CD56⁺ cells primes monocytes for IL-12p70 production in response to LPS.

<p>1 ·10⁵ CD14⁺ monocytes were stimulated with 100 ng/ml LPS in combination with 2.5 ng/ml IFN-γ, supernatants of unstimulated CD56⁺ cells, supernatants of IL-23 plus IL-1β stimulated CD56⁺ cells, or medium alone for 24 hours (A). 1 ·10⁵ CD14⁺ monocytes were stimulated with 100 ng/ml LPS in combination supernatants of IL-23 plus IL-1β stimulated CD56+ cells, plus or minus 2 µg/ml anti-IFN-γ for 24 hours (B). Supernatants were taken and IL-12p70 production was measured by ELISA. Data are means±SD of triplicates from one representative experiment of three.</p

GM-CSF and IFN-γ prime monocytes for enhanced IL-23 production.

<p>1 ·10⁵ CD14⁺ monocytes were prestimulated with GM-CSF (A) or IFN-γ (B) for 16 hours in concentrations indicated. Subsequently cells were stimulated with heat killed Salmonella (moi = 10) or left unstimulated for 24 hours. Supernatants were taken and IL-23 production was measured by ELISA. Data are means±SD of triplicates in one representative experiment of five.</p

Supernatants of <i>Salmonella</i>-infected Mφ1 induce IFN-γ production in primary human CD56⁺ cells.

<p>(A) IFN-γ production by CD56⁺ NK/NK-like T cells from two donors. Cells were cultured for 48 hours in the presence of supernatants obtained from uninfected or <i>Salmonella</i> infected Mφ1 (from two donors) with or without additional exogenous recombinant IL-18. As a control, medium only was used. Data are means±SD of triplicates in one representative experiment of three. (B) IFN-γ production in CD56⁺ cells induced by supernatants of <i>Salmonella</i>-infected Mφ1 is blocked by neutralizing IL-12p40 or IL-1β, but not by neutralizing IL-18. CD56⁺ NK/NK-like T cells were cultured for 48 hours in the presence or absence of supernatant obtained from Mφ1 cells infected with <i>Salmonella</i>, in the presence or absence of an IL-12p40, IL-18 or IL-1β neutralizing antibody. IFN-γ concentration was determined by ELISA. IFN-γ production induced by supernatants of <i>Salmonella</i> infected Mφ1 is set at 100%. A paired two-tailed student's t-test was used for statistical analysis. Data are means±SEM of experiments with cells obtained from 10 different donors.</p