Responsiveness of blood and sputum inflammatory cells in Japanese COPD patients, non-COPD smoking controls, and non-COPD nonsmoking controls

Figures & data

Table 1 Summary of subject demographics and baseline characteristics

Demographics and measurements	Non-COPD NSC group (N=10)	Non-COPD SC group (N=10)	COPD group (N=10)
Demographics
Age, years	69.4±5.2	64.8±7.8	62.2±6.6
Sex, males/females	5/5	9/1	7/3
Height, cm	158.3±4.3	165.7±7.0	159.9±6.8
Weight, kg	59.3±11.2	63.0±10.5	48.9±7.3
Smoking history
Current/former/never	0/0/10	8/2/0	3/7/0
Number of pack-years	0	44.3±35.0	65.4±30.2
Spirometry
FVC, L	3.1±0.9a	3.8±0.9b	2.8±0.8
FEV1, L	2.4±0.7c	2.9±0.7d	1.2±0.9
FEV1, %predicted	79.5±4.2d	76.6±5.8c	40.4±17.6
FEV1/FVC, %	79.5±4.2d	77.0±5.6c	39.6±17.9

Notes:

a P<0.05,

b P<0.01,

c P<0.005, and

d P<0.0001 (difference between COPD group and non-COPD NSC group/non-COPD SC group, by Dunnett’s test).

Abbreviations: COPD, chronic obstructive pulmonary disease; FEV₁, forced expiratory volume in 1 second; FVC, forced vital capacity; SC, smoking controls; NSC, nonsmoking controls.

Table 2 Summary of the differences in biomarker levels before and after stimulation in PMBC and induced sputum cells (all subjects)

Inflammatory biomarkers	Non-COPD NSC group (N=10)	Non-COPD SC group (N=10)	COPD group (N=10)
PBMC: Mean ± SD difference (pre- and poststimulation with TNF-α)
IL-8, pg/mL	3,520.3±2,979.1	2,630.0±1,977.2	4,147.0±2,974.5
IL-6, pg/mL	41.4±59.7	−36.7±92.2	40.7±85.7
MMP-9, pg/mL	268.3±442.2	443.9±679.4	2,633.2±4,630.7
PBMC: Mean ± SD difference (pre- and poststimulation with LPS)
IL-8, pg/mL	46,881.7±24,615.3	34,948.2±9,743.3	34,192.7±15,365.8
IL-6, pg/mL	7,309.7±5,511.0	7,979.4±3,285.2	8,393.5±3,200.5
MMP-9, pg/mL	292.3±508.3	446.0±697.3	2,997.4±5,267.0
TNF-α, pg/mL	1,296.0±841.6	4,419.9±4,421.3	8,146.9±20,195.6
Sputum: Mean ± SD difference (pre- and poststimulation with TNF-α)
IL-8, pg/mL	−9,191.8±22,715.9	−619.1±17,886.7	2,714.0 ± 11,359.2
IL-6, pg/mL	−446.4±1,223.5	556.3±2,855.3	62.1±716.4
MMP-9, pg/mL	−801.4±2,643.3	−726.9±1,971.3	−311.3±974.9
Sputum: Mean ± SD difference (pre- and poststimulation with LPS)
IL-8, pg/mL	20,785.4±48,223.6	1,175.8±2,180.1	2,885.3±15,438.1
IL-6, pg/mL	1,702.2±2,549.0	−45.3±173.5	1,496.9±2,615.2
MMP-9, pg/mL	−1,011.5±2,926.7	−39.0±321.4	132.6±2,715.0
TNF-α, pg/mL	9,773.4±26,966.8	2,533.8±14,710.9	3,589.3±8,590.8

Abbreviations: COPD, chronic obstructive pulmonary disease; IL-8, interleukin-8; IL-6, interleukin-6; LPS, lipopolysaccharide; MMP-9, matrix metalloproteinase-9; SC, smoking controls; NSC, nonsmoking controls; PBMC, peripheral blood mononuclear cells; SD, standard deviation; TNF-α, tumor necrosis factor-alpha.

Figure 1 Basal release of TNF- α (pg/mL) from PBMCs in patients with COPD, non-COPD SC, and non-COPD NSC.

Notes: Data are presented as mean ± standard deviation; n=10 in each group. ^aDifference between COPD and non-COPD NSC groups by Dunnett’s test.

Abbreviations: TNF-α, tumor necrosis factor-alpha; LPS, lipopolysaccharide; COPD, chronic obstructive pulmonary disease; SC, smoking controls; NSC, nonsmoking controls; PBMCs, peripheral blood mononuclear cells.

Figure 2 LPS-induced TNF-α release (pg/mL) from PBMC (A) and sputum cells (B) in patients with COPD, non-COPD SC, and non-COPD NSC.

Notes: Data are presented as mean ± standard deviation; N=10 in each group.
Abbreviations: TNF-α, tumor necrosis factor-alpha; LPS, lipopolysaccharide; COPD, chronic obstructive pulmonary disease; SC, smoking controls; NSC, nonsmoking controls; PBMC, peripheral blood mononuclear cells.

Table 3 Inflammatory cell counts in sputum

Inflammatory cells	Sputum cell count
	Non-COPD NSC group (N=10)	Non-COPD SC group (N=10)	COPD group (N=10)
Total cell count, ×10^5 cells/mL	8,433.0±4,822.1	19,027.6±20,716.7	26,059.1±48,773.9
Neutrophils, ×10^5 cells/mL	7,235.9±4,727.2	17,573.1±20,508.7	24,528.6±48,376.7
Macrophages, ×10^5 cells/mL	1,029.4±780.7	1,191.8±725.7	831.7±775.1
Lymphocytes, ×10^5 cells/mL	0.7±0.8	0.5±0.5	0.6±1.0
Eosinophils, ×10^5 cells/mL	0.9±1.3	2.2±5.2	6.4±12.2

Note: All data are expressed as mean ± standard deviation.

Abbreviations: COPD, chronic obstructive pulmonary disease; SC, smoking controls; NSC, nonsmoking controls.

Table 4 Correlations between sputum inflammatory cell counts and sputum biomarkers in all subjects

Inflammatory cells	Sputum
	IL-8	IL-6	MMP-9	TNF-α
Absolute number
Total cells	−0.28	−0.27	0.20	−0.10
Neutrophils	−0.34	−0.31	0.22	−0.13
Macrophages	0.56c	0.48b	0.19	0.49b
Cell differentiation
Percentage of neutrophils	−0.69e	−0.63d	0.18	−0.41a
Percentage of macrophages	0.72e	0.64d	−0.12	0.49b

Notes:

a P<0.05,

b P<0.01,

c P<0.005,

d P<0.0005, and

e P<0.0001 (correlation between variables, by Spearman’s rank correlation coefficient).

Abbreviations: IL-8, interleukin-8; IL-6, interleukin-6; MMP-9, matrix metalloproteinase-9; TNF-α, tumor necrosis factor-alpha.

Table S1 Summary of biomarkers in peripheral blood mononuclear cells (stimulated by TNF-α or LPS)

Inflammatory cells	Non-COPD NSC group (N=10)		Non-COPD SC group (N=10)		COPD group (N=10)
	(−)TNF-α	(+)TNF-α	(−)TNF-α	(+)TNF-α	(−)TNF-α	(+)TNF-α
IL-8, pg/mL	421.8±361.5	3,942.1±3,293.4	525.5±360.0	3,155.5±1,796.6	315.8±155.0	4,462.8±3,087.4
IL-6, pg/mL	3.0±4.7	44.4±63.1	50.6±89.2	13.9±12.0	3.6±6.6	44.25±91.9
MMP-9, pg/mL	500.0±0.0b	768.3±442.2	500.0±0.0b	943.9±679.4	500.0±0.0b	3,133.2±4,630.7
	(−)LPS	(+)LPS	(−)LPS	(+)LPS	(−)LPS	(+)LPS
IL-8, pg/mL	421.8±361.5	47,303.5±24,858	525.5±360.0	35,473.7±9,802.1	315.8±155.0	34,508.5±15,412.7
IL-6, pg/mL	3.0±4.7	7,312.7±5,512.9	50.6±89.2	8,030.0±3,340.4	3.6±6.6	8,397.1±3,203.8
MMP-9, pg/mL	500.0±0.0b	792.3±508.3	500.0±0.0b	946.0±697.3	500.0±0.0b	3,497.4±5,267.0
TNF-α, pg/mL	9.5±5.2a	1,305.4±840.7	80.3±95.7	4,500.2±4,453.8	81.6±111.4	8,228.5±20,171.3

Notes: All data are expressed as mean ± standard deviation.

a P<0.05 (difference between COPD groups and non-smokers, by the Dunnett’s test); (+) with stimulation, (−) without stimulation.

b The levels of MMP-9 before stimulation with TNF-α were below the limit of detection (1,000 pg/mL).

Abbreviations: COPD, chronic obstructive pulmonary disease; IL-8, interleukin-8; IL-6, interleukin-6; LPS, lipopolysaccharide; MMP-9, matrix metalloproteinase-9; SC, smoking controls; NSC, nonsmoking controls; TNF-α, tumor necrosis factor-alpha.