Effects of budesonide on toll-like receptor expression in alveolar macrophages from smokers with and without COPD

Figures & data

Table 1 Characteristics of the participants

	Healthy nonsmokers	Smokers without COPD	Smokers with COPD
Subjects, n	10	11	10
Age (years), mean (range)	54 (41–70)	55 (41–64)	60 (48–73)
Sex (male/female)	7/3	6/5	2/8
Smoking (pack-years), mean (range)	0	38 (20–60)	34 (20–60)
FEV1 % predicted (post-bronchodilator)	99.4 (91.96–106.84)	93.09* (87.47–98.72)	57.7***,### (46.47–68.93)
FEV1/FVC (post-bronchodilator)	79.1 (74.67–83.53)	75.45 (70.19–80.72)	56.1***,### (51.71–60.49)
Numbers of total cells in the BAL fluid (10^6 cells)	12.7 (8.8–16.8)	40.7** (26.5–51.0)	29.8** (27.7–40.9)
Proportion of macrophages in the BAL fluid (%)	93.3 (87.3–94.6)	94 (90.6–95.3)	89.65 (83.6–95.6)

Notes: Age, smoking history and lung function index are presented as mean values (95% CI). Numbers of total cells in the BAL fluid and proportion of macrophages in the BAL fluid are presented as median values (25th–75th percentile). Between-group comparisons were assessed by Kruskal–Wallis test with the Mann–Whitney U-test as a post hoc test.

*, **, and *** indicate P<0.05, P<0.01, and P<0.001, respectively, compared with the healthy nonsmokers.

### Indicates P<0.001 compared with the smokers without COPD.

Abbreviations: FEV₁, forced expiratory volume in 1 second; FVC, forced vital capacity; BAL, bronchoalveolar lavage.

Figure 1 Secretion of IL-8 in the BAL fluid AMs from the healthy controls and smokers with and without COPD incubated for 6 hours in control medium (A), LPS (B), PGN (C), and TNF-α (D) in the absence and presence of Bud.

Notes: Data are presented as median values and interquartile ranges. ^# and ^##, respectively, indicate P<0.05 and P<0.01 for the effect of different stimuli (medium vs LPS/PGN/TNF-α). * and **, respectively, indicate P<0.05 and P<0.01 for the effect of Bud (LPS/PGN/TNF-α vs LPS + Bud/PGN + Bud/TNF-α + Bud). ^¤Indicates P<0.05 for the effect of PGN (Bud vs PGN + Bud). ^& and ^&&, respectively, indicate P<0.05 and P<0.01 compared with the healthy control cells. Different designators used for comparisons within groups and between groups.
Abbreviations: IL-8, interleukin-8; BAL, bronchoalveolar lavage; AM, alveolar macrophage; LPS, lipopolysaccharide; PGN, peptidoglycan; TNF-α, tumor necrosis factor-alpha; Bud, budesonide.

Figure 2 Secretion of TNF-α in the BAL fluid AMs from healthy controls and smokers with and without COPD incubated for 6 hours in the control medium (A), LPS (B), and PGN (C) in the absence and presence of Bud.

Notes: Data are presented as median values and interquartile ranges. ^#Indicates P<0.05 for the effect of different stimuli (medium vs LPS/PGN). *Indicates P<0.05 for the effect of Bud (LPS/PGN vs LPS + Bud/PGN + Bud). ^¤Indicates P<0.05 for the effect of LPS/PGN (Bud vs LPS + Bud/PGN + Bud). Different designators used for comparisons within groups and between groups.
Abbreviations: TNF-α, tumor necrosis factor-alpha; BAL, bronchoalveolar lavage; AM, alveolar macrophage; LPS, lipopolysaccharide; PGN, peptidoglycan; Bud, budesonide.

Figure 3 Expression of TLR2 mRNA in the BAL fluid AMs from the healthy controls and smokers with and without COPD incubated for 6 hours in the control medium (A), LPS (B), PGN (C), and TNF-α (D) in the absence and presence of Bud.

Notes: Data are presented as median values and interquartile ranges. *Indicates P<0.05 for the effect of Bud (medium/LPS/PGN vs Bud/LPS + Bud/PGN + Bud). ^¤Indicates P<0.05 for the effect of PGN (Bud vs PGN + Bud). Different designators used for comparisons within groups and between groups.
Abbreviations: TLR2, toll-like receptor 2; mRNA, messenger RNA; BAL, bronchoalveolar lavage; AM, alveolar macrophage; LPS, lipopolysaccharide; PGN, peptidoglycan; TNF-α, tumor necrosis factor-alpha; Bud, budesonide.

Figure 4 Expression of TLR4 mRNA in the BAL fluid AMs from the healthy controls and smokers with and without COPD incubated for 6 hours in the control medium (A), LPS (B), PGN (C) and TNF-α (D) in the absence and presence of Bud.

Notes: Data are presented as median values and interquartile ranges. ^#Indicates P<0.05 for the effect of different stimuli (medium vs LPS/TNF-α). ^¤Indicates P<0.05 for the effect of LPS/PGN (Bud vs LPS + Bud/PGN + Bud). Different designators used for comparisons within groups and between groups.
Abbreviations: TLR4, toll-like receptor 4; mRNA, messenger RNA; BAL, bronchoalveolar lavage; AM, alveolar macrophage; LPS, lipopolysaccharide; TNF-α, tumor necrosis factor-alpha; PGN, peptidoglycan; Bud, budesonide.

Figure 5 Inhibition of IL-8 secretion in response to PGN (A–C) or LPS (D–F) stimulation by Bud after preincubation without or with TLR2/TLR4 antibodies from the healthy controls and smokers with and without COPD.

Notes: Values are presented as percentage of IL-8 secretion stimulated by PGN (A–C) or LPS (D–F). Data are presented as median values and interquartile ranges.
Abbreviations: IL-8, interleukin-8; PGN, peptidoglycan; LPS, lipopolysaccharide; TLR2, toll-like receptor 2; TLR4, toll-like receptor 4; Bud, budesonide.