Chest
Volume 123, Issue 4, April 2003, Pages 1240-1247
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Laboratory and Animal Investigations
Neutrophil Chemotactic Activity of Sputum From Patients With COPD: Role of Interleukin 8 and Leukotriene B4

https://doi.org/10.1378/chest.123.4.1240Get rights and content

Study objectives:

Neutrophilic inflammation is a major feature of COPD. Several factors in bronchial secretions have been identified as chemoattractants for neutrophils. The present study was designed to assess the contribution of interleukin (IL)-8 and leukotriene B4 (LTB4) to neutrophil chemotaxis evoked by sputum obtained from patients with established COPD.

Design:

Sputum supernatant of 20 patients with COPD was used as chemoattractant in a 96-well chemotaxis chamber, with subsequent quantification of migrated cells by a luminescence assay. The contribution of IL-8 and LTB4 to chemotaxis was determined by addition of a neutralizing antibody and a selective receptor antagonist, respectively.

Measurements and results:

COPD sputum caused neutrophil chemotaxis in a concentration-dependent manner, with a maximum response evoked with a 10-fold dilution of the original sample. Pretreatment of sputum or neutrophils with either an anti–IL-8 antibody or the LTB4 antagonist, SB 201146, led to a concentration-dependent inhibition of sputum-induced neutrophil chemotaxis, with a maximum suppression (mean ± SEM) of 29.2 ± 4.9% (p < 0.001) from baseline by 100 ng/mL of anti–IL-8 antibody, and 45.6 ± 7% (p < 0.02) by 10 μmol/L of SB 201146. The combination of the anti–IL-8 antibody and SB 201146 inhibited neutrophil chemotaxis, but this was not significantly greater than the effect of SB 201146 or anti–IL-8 alone.

Conclusions:

These data confirm the importance of IL-8 and LTB4 as chemoattractants for neutrophils in bronchial secretions from patients with COPD, and suggest that specific inhibitors may have therapeutic potential in COPD.

Section snippets

Patients

Twenty patients with diagnosed COPD underwent sputum induction. All patients fulfilled the diagnostic criteria of the British Thoracic Society.22 None of the patients had actively smoked for at least 6 months prior to the study; received inhaled or oral corticosteroids, or other systemic drugs including theophylline, antibiotics or nonsteroidal anti-inflammatory drugs; or reported any acute exacerbations for at least 4 weeks prior to sputum induction. The study was approved by local regulatory

Concentration of Chemoattractants in Pooled Sputum Supernatant

The concentration of IL-8 and LTB4 in the pooled sputum supernatant was 20.9 ng/mL and 12.6 pg/mL, respectively.

IL-8 and LTB4-Induced Neutrophil Chemotaxis

Under resting conditions or after exposure to diluent, human neutrophils underwent spontaneous chemotaxis affecting 0.4 ± 0.2% of the total cell population after 1 h. The addition of IL-8 and LTB4 augmented chemotaxis over the spontaneous response in a concentration-dependent manner with the effective concentration causing a 50% fall (EC50) values of 4 ng/mL and 3.3 nmol, respectively

Discussion

Neutrophilic inflammation is an important feature of COPD and may contribute to mucus hypersecretion and destruction of alveoli. Several factors have been identified as chemoattractants for peripheral blood neutrophils, which may result in an increased trafficking of these cells to bronchial tissues. The present study was designed to assess the contribution of IL-8 and LTB4 present in the sputum of patients with established COPD on sputum-induced neutrophil chemotaxis.

The methodology was

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