Specific IgA and metalloproteinase activity in bronchial secretions from stable chronic obstructive pulmonary disease patients colonized by Haemophilus influenzae

Laura Millares, Alicia Marin, Judith Garcia-Aymerich, Jaume Sauleda, José Belda, Eduard Monsó, PAC-COPD Study Group, Josep M Antó, Judith Garcia-Aymerich, Marta Benet, Jordi de Batlle, Ignasi Serra, David Donaire-Gonzalez, Stefano Guerra, Joaquim Gea, Eva Balcells, Angel Gayete, Mauricio Orozco-Levi, Ivan Vollmer, Joan Albert Barberà, Federico P Gómez, Carles Paré, Josep Roca, Robert Rodriguez-Roisin, Xavier Freixa, Diego A Rodriguez, Elena Gimeno, Karina Portillo, Jaume Ferrer, Jordi Andreu, Esther Pallissa, Esther Rodríguez, Pere Casan, Rosa Güell, Ana Giménez, Eduard Monsó, Alicia Marín, Sara Barea, Josep Morera, Eva Farrero, Joan Escarrabill, Antoni Ferrer, Jaume Sauleda, Alvar G Agustí, Bernat Togores, Juan Bautista Gáldiz, Lorena López, José Belda, Laura Millares, Alicia Marin, Judith Garcia-Aymerich, Jaume Sauleda, José Belda, Eduard Monsó, PAC-COPD Study Group, Josep M Antó, Judith Garcia-Aymerich, Marta Benet, Jordi de Batlle, Ignasi Serra, David Donaire-Gonzalez, Stefano Guerra, Joaquim Gea, Eva Balcells, Angel Gayete, Mauricio Orozco-Levi, Ivan Vollmer, Joan Albert Barberà, Federico P Gómez, Carles Paré, Josep Roca, Robert Rodriguez-Roisin, Xavier Freixa, Diego A Rodriguez, Elena Gimeno, Karina Portillo, Jaume Ferrer, Jordi Andreu, Esther Pallissa, Esther Rodríguez, Pere Casan, Rosa Güell, Ana Giménez, Eduard Monsó, Alicia Marín, Sara Barea, Josep Morera, Eva Farrero, Joan Escarrabill, Antoni Ferrer, Jaume Sauleda, Alvar G Agustí, Bernat Togores, Juan Bautista Gáldiz, Lorena López, José Belda

Abstract

Background: Haemophilus influenzae is the most common colonizing bacteria of the bronchial tree in chronic obstructive pulmonary disease (COPD), and positive cultures for this potentially pathogenic microorganism (PPM) has been associated with local inflammation changes that may influence the relationships between H. influenzae and the bronchial mucosa.

Methods: A cross-sectional analysis of stable COPD patients enrolled in the Phenotype and Course of Chronic Obstructive Pulmonary Disease (PAC-COPD) Study, focusing on bronchial colonization by H. influenzae, was performed. Specific IgA against the PPM was measured by optical density, and metalloproteinase-9 (MMP-9) and tissue inhibitor of metalloproteinase-1 (TIMP-1) using ELISA in sputum samples. Levels in patients colonized by H. influenzae and non-colonized patients were compared.

Results: Sputum supernatant for the measurement of specific IgA against H. influenzae was available from 54 stable COPD patients, who showed levels of specific IgA significantly lower in colonized (n=21) than in non-colonized patients (n=33) (15 [4-37] versus 31 [10-75], p=0.033, Mann-Whitney U test). Proenzyme MMP-9 was measured in 44 patients, and it was higher in colonized (n=12, 1903 [1488-6699] ng/ml) than in non-colonized patients (n=32, 639 [373-972] ng/ml) (p<0.001, Mann-Whitney U test). Active form of MMP-9 was also higher in colonized (126 [25-277] ng/ml) than in non-colonized patients (39 [14-68] ng/ml) (p=0.021, Mann-Whitney U test), and the molar ratio between proenzyme MMP-9 and TIMP-1 was above 1 (2.1 [0.1-12.5]) in colonized patients, significantly higher than the ratio found in non-colonized patients (0.2 [0.08-0.5]) (p=0.030, Mann-Whitney U test).

Conclusions: Clinically stable COPD patients colonized by H. influenzae had lower levels of specific IgA against the microorganism and higher values of the active form of MMP-9 in their sputum supernatant than non-colonized patients. Bronchial colonization by H. influenzae may cause structural changes in the extracellular matrix through a defective defense and the production of active metalloproteinases.

Figures

Figure 1
Figure 1
Ten distinct isolates of H. Influenzae, isolated from COPD patients, used as capture antigen in ELISA testing.
Figure 2
Figure 2
Flow chart showing the number of subjects used in each analysis.
Figure 3
Figure 3
Level of specific IgA against H. influenzae in patients colonized by this bacteria (n=21) and non-colonized patients (n=33). Results expressed as optical density (OD) ratio between patients and healthy controls.
Figure 4
Figure 4
Proenzyme MMP-9 concentration in the sputum supernatant of patients colonized by H. influenzae (n=12) and non-colonized patients (n=32).
Figure 5
Figure 5
TIMP-1 concentration in the sputum supernatant of patients colonized by H. influenzae (n=12) and non-colonized patients (n=32).
Figure 6
Figure 6
Active MMP-9 concentration in the sputum supernatant of patients colonized by H. influenzae (n=12) and non- colonized patients (n=32).
Figure 7
Figure 7
Molar ratio between proenzyme MMP-9 and TIMP-1 in colonized patients colonized by H. influenzae (n=12) and non-colonized patients (n=32).

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