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This study investigates the effect of autolysed wild-type (WT) pneumococci on the production of pro-inflammatory cytokines TNF and IFN-γ in peripheral blood mononuclear cells (PBMC) stimulated by intact D39 pneumococci. PBMC from six donors were exposed to varying concentrations of autolysed bacteria alongside intact bacteria. Cytokine levels (TNF, IL-6, IL-8, IL-10, and IFN-γ) were measured after 24 hours or five days. Results indicate that autolysed WT pneumococci dose-dependently inhibit cytokine production in a statistically significant manner.
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TNF IFN-γ * * * ** * 150 6 100 ng/ml ng/ml 4 50 2 0 0 +WT 3×107/ml AL- 3×107/ml +WT 3×106/ml +WT 3×105/ml +WT 3×107/ml AL- 3×107/ml +WT 3×106/ml +WT 3×105/ml IL-6 IL-8 IL-10 * * 2 300 150 1.5 200 ng/ml ng/ml 100 ng/ml 1 100 50 0.5 0 0 0 +WT 3×107/ml +WT 3×107/ml +WT 3×107/ml AL- 3×107/ml +WT 3×106/ml +WT 3×105/ml AL- 3×107/ml +WT 3×106/ml +WT 3×105/ml AL- 3×107/ml +WT 3×106/ml +WT 3×105/ml FIG. S3. FIG. S3. Presence of autolysed WT pneumococci, dose-dependently inhibit production of TNF and IFN-γ in response to AL- intact pneumococci. PBMC from six blood donors were stimulated intact D39 AL- at 3 × 107 bacteria/ml mixed with 3 × 105, 3 × 106 and 3 × 107 bacteria/ml (before autolysis) of autolysed D39 WT. Cytokines were measured in the supernatant after 24 h (TNF, IL-6, IL-8 and IL-10) or 5 d (IFN-γ). Medians and quartiles are shown by boxes and the bars represent the 10th and 90th percentiles, respectively. The response to AL- bacteria with and without addition of WT bacteria within the same donor was compared using Wilcoxon matched pairs test. * p < 0.05; ** p < 0.01.