Figures
- FIGURE 1
Non-typeable Haemophilus influenzae (NTHi)-stimulated cytokine production. Peripheral blood mononuclear cells from healthy control children (n=17), children with protracted bacterial bronchitis (PBB) (n=19) and children with bronchiectasis (BE) (n=20) were cultured ex vivo in the presence of NTHi. Supernatant was collected at 24 h for the innate response cytokines (interleukin (IL)-1β, interleukin-1 receptor antagonist (IL-1Ra), IL-18 and IL-6) and at 72 h for the adaptive response cytokines (interferon (IFN)-γ and IL-10). a) IL-1β concentration. b) IL-1Ra concentration. c) IL-18 concentration. d) IL-6 concentration. e) IFN-γ concentration. f) IL-10 concentration. Box and whisker plots display median, interquartile ranges and range. **: p<0.01 by Wilcoxon matched-pairs signed rank test; ***: p<0.001 by Wilcoxon matched-pairs signed rank test.
- FIGURE 2
Effects of interleukin-1 receptor antagonist (IL-1Ra) on peripheral blood mononuclear cell (PBMC) responses to non-typeable Haemophilus influenzae (NTHi). PBMCs were cultured ex vivo with NTHi in the presence or absence of the IL-1Ra anakinra. Data are from 14–15 healthy control children, 17–19 children with protracted bacterial bronchitis (PBB) and 12–13 children with bronchiectasis (BE). a) Interleukin (IL)-1β concentration. b) IL-6 concentration. c) IL-18 concentration. d) IFN-γ concentration. e) IL-10 concentration. ns: nonsignificant. *: p<0.05 by Wilcoxon matched-pairs signed rank test; **: p<0.01 by Wilcoxon matched-pairs signed rank test; ***: p<0.001 by Wilcoxon matched-pairs signed rank test.
- FIGURE 3
Rhinovirus (RV)1B-stimulated cytokine production. Peripheral blood mononuclear cells from children with protracted bacterial bronchitis (PBB) were cultured ex vivo in the presence of RV1B. Supernatant was collected at 24 h for the innate response cytokines (interleukin (IL)-1β, interleukin-1 receptor antagonist (IL-1Ra), IL-18 and IL-6) and at 72 h for the adaptive response cytokines (interferon (IFN)-γ and IL-10). a) IL-1β concentration (n=10). b) IL-1Ra concentration (n=10). c) IL-18 concentration (n=6). d) IL-6 concentration (n=10). e) IFN-γ concentrations (n=8). f) IL-10 concentration (n=8). Box and whisker plots display median, interquartile ranges and range. *: p<0.05 by Wilcoxon matched-pairs signed rank test; **p<0.01 by Wilcoxon matched-pairs signed rank test.
- FIGURE 4
Effects of interleukin-1 receptor antagonist (IL-1Ra) on peripheral blood mononuclear cell (PBMC) responses to rhinovirus (RV)1B. PBMCs from children with protracted bacterial bronchitis (PBB) were cultured ex vivo with RV1B in the presence or absence of the IL-1Ra anakinra. a) Interleukin (IL)-1β concentration (n=11). b) IL-6 concentration (n=11). c) Interferon (IFN)-γ concentration (n=12). d) IL-10 concentration (n=12). *: p<0.05 by Wilcoxon matched-pairs signed rank test.
- FIGURE 5
Effects of interleukin (IL)-1β on peripheral blood mononuclear cell (PBMC) responses to non-typeable Haemophilus influenzae (NTHi) and rhinovirus (RV)1B. PBMCs from healthy adults were cultured with NTHi or RV1b in the presence or absence of recombinant IL-1β. Data are from 7–14 experiments. a) IL-6 concentration (n=7). b) Interferon (IFN)-γ concentration. c) IL-10 concentration. Mean and standard deviations are shown. *: p<0.05 by Wilcoxon matched-pairs signed rank test; ***: p<0.001 by Wilcoxon matched-pairs signed rank test.
- FIGURE 6
Interleukin-1 blockade effects interferon (IFN)-γ production by multiple cell types. a) Gating strategy for flow cytometry: CD56+ natural killer (NK) cells (CD3−CD56+), CD3+ T-cells (CD3+CD56−) and other cells (CD3−CD56−) were identified within the total gated lymphocytes. The percentage of IFN-γ-producing cells was then evaluated in each of the cell subtypes. b) Percentage of non-typeable Haemophilus influenzae (NTHi)-stimulated IFN-γ-producing cells in the absence and presence of interleukin-1 receptor antagonist (IL-1Ra) at 24 h post-stimulation (n=9). *: p<0.05 by Wilcoxon matched-pairs signed rank test; **: p<0.01 by Wilcoxon matched-pairs signed rank test.
Tables
- TABLE 1
Subject demographics and full blood counts
Controls# Recurrent PBB# Recurrent PBB¶ Bronchiectasis# Subjects n 20 20 13 20 Age years 2.2 (0.3–4.5) 1.6 (0.9–2.8) 3.2 (1.6–6.1) 3.1 (2.1–5.0) Boys n 15 (75%) 16 (80%) 9 (70%) 14 (70%) Cough score NA 3 (1–3) 3 (1–4) 2 (1–3) Full blood count ×106 L−1 Total cell count 8.90 (8.18–10.2) 11.0 (9.05–12.2) 9.30 (8.25–10.3) 9.60 (7.70–11.8) Neutrophils 2.88 (1.69–3.72) 3.63 (2.49––4.96) 2.55 (2.13–3.09) 2.96 (2.44–4.26) Monocytes 0.80 (0.59–1.02) 1.09 (0.78–1.16) 0.71 (0.63–0.94) 0.84 (0.69–0.96) Eosinophils 0.34 (0.12–0.67) 0.38 (0.19–0.59) 0.22 (0.17–0.67) 0.35 (0.24–0.74) Lymphocytes 4.82 (3.81–5.87) 5.32 (4.04–6.60) 4.97 (3.99–5.97) 4.51 (3.83–5.81) Data are presented as median (interquartile range) unless otherwise stated. PBB: protracted bacterial bronchitis; IQR: interquartile range; NA: not applicable. #: figures 1 and 2; ¶: figures 3 and 4.
Supplementary Material
Please note: supplementary material is not edited by the Editorial Office, and is uploaded as it has been supplied by the author.
Supplementary materials and methods, table S1, and figure S1 00025-2017_Suppl
Supplementary Material
K.J. Baines 00025-2017_Baines
A.B. Chang 00025-2017_Chang
P.G. Gibson 00025-2017_Gibson
J.W. Upham 00025-2017_Upham
