Remodeling associated expression of matrix metalloproteinase 9 but not tissue inhibitor of metalloproteinase 1 in airway epithelium: Modulation by immunostimulatory DNA

doi:10.1016/j.jaci.2005.12.1324

Journal of Allergy and Clinical Immunology

Volume 117, Issue 3, March 2006, Pages 618-625

https://doi.org/10.1016/j.jaci.2005.12.1324 Get rights and content

Background

Matrix metalloproteinase 9 (MMP-9) and its tissue inhibitor of metalloproteinase 1 (TIMP-1) are hypothesized to play a role in the pathogenesis of airway remodeling in asthma.

Objective

We have used a mouse model of airway remodeling to determine the pattern of expression of MMP-9 and TIMP-1 in airway epithelium and peribronchial cells, and assess whether TIMP-1, an inhibitor of MMP-9, is expressed at the same sites in the airway. In addition, we have investigated whether immunostimulatory sequences (ISSs) of DNA modulate levels of expression of MMP-9, TIMP-1, and peribronchial fibrosis.

Methods

Levels of lung MMP-9 and TIMP-1 were assessed by zymography, ELISA, and immunohistochemistry.

Results

Repetitive ovalbumin challenge induced a significant increase in levels of MMP-9, TIMP-1, and peribronchial collagen deposition. The pattern of expression of MMP-9 and TIMP-1 in the remodeled airway was significantly different. MMP-9 but not TIMP-1 was expressed in airway epithelium, whereas both MMP-9 and TIMP-1 were expressed in peribronchial inflammatory cells. ISS significantly reduced expression of MMP-9 in airway epithelium (which immunostained positive for Toll receptor 9), as well as in peribronchial inflammatory cells. In vitro studies demonstrated that ISS inhibited bone marrow macrophage generation of MMP-9.

Conclusion

Allergen-induced peribronchial fibrosis is associated with expression of MMP-9 and TIMP-1 at different anatomical sites in the remodeled airway. The ability of ISS to inhibit the expression of MMP-9 in airway epithelium (a site where its inhibitor TIMP-1 is not induced by allergen challenge) may be important in determining whether ISS contributes to reductions in airway remodeling by reducing levels of MMP-9.

Clinical implications

Immunostimulatory sequences of DNA, which are being investigated as novel therapeutics in asthma, inhibit airway remodeling in mice as well as epithelial expression of MMP-9, an enzyme that degrades the extracellular matrix proteins surrounding the airway.

Section snippets

Induction of allergen-induced airway remodeling

The methods we have used to administer ovalbumin allergen to induce airway remodeling in mice have previously been described.16, 17, 18 In brief, BALB/c mice (16 mice/group; Jackson Laboratory, Bar Harbor, Me) were used when they reached 8 to 10 weeks of age. Mice were immunized subcutaneously on days 0, 7, 14, and 21 with 25 μg ovalbumin (grade V; Sigma-Aldrich, St Louis, Mo) adsorbed to 1 mg alum (Sigma-Aldrich) in 200 μL normal saline. Intranasal ovalbumin challenges (20 ng/50 μL in PBS)

Immunohistochemical detection of epithelial cell vs peribronchial cells expressing MMP-9 and TIMP-1 in the remodeled airway: Effect of ISS

Minimal peribronchial expression of either MMP-9 (Fig 1, A) or TIMP-1 (Fig 1, D) was noted in nonovalbumin-challenged mice by immunohistochemistry. In contrast, both MMP-9 (Fig 1, B) and TIMP-1 (Fig 1, E) were significantly expressed after repetitive ovalbumin challenge for 3 months in the remodeled airway. However, the peribronchial distribution of MMP-9 expression differed from that of TIMP-1 expression in the remodeled airway in that MMP-9 was expressed in airway epithelium (Fig 1, B)

Discussion

In this study, we have demonstrated that allergen induced airway remodeling is associated with significantly increased levels of expression of MMP-9 and its inhibitor TIMP-1. Interestingly, immunostaining of lungs derived from mice with remodeled airways demonstrated a distinctly different pattern of MMP-9 expression compared with TIMP-1 expression in the remodeled airway, with MMP-9, but not TIMP-1, expressed in airway epithelium. Thus, the localized absence of expression of TIMP-1, an

References (25)

M. Hoshino et al.
Bronchial subepithelial fibrosis and expression of matrix metalloproteinase-9 in asthmatic airway inflammation
J Allergy Clin Immunol
(1998)
H. Tanaka et al.
Sputum matrix metalloproteinase-9: tissue inhibitor of metalloproteinase-1 ratio in acute asthma
J Allergy Clin Immunol
(2000)
M. Hoshino et al.
Inhaled corticosteroids decrease subepithelial collagen deposition by modulation of the balance between matrix metalloproteinase-9 and tissue inhibitor of metalloproteinase-1 expression in asthma
J Allergy Clin Immunol
(1999)
D.D. Cataldo et al.
Matrix metalloproteinase-9 deficiency impairs cellular infiltration and bronchial hyperresponsiveness during allergen-induced airway inflammation
Am J Pathol
(2002)
W.C. Parks et al.
Matrix metalloproteinases in lung biology
Respir Res
(2001)
H. Lemjabbar et al.
Contribution of 92 kDa gelatinase/type IV collagenase in bronchial inflammation during status asthmaticus
Am J Respir Crit Care Med
(1999)
A.M. Vignola et al.
Sputum metalloproteinase-9/tissue inhibitor of metalloproteinase-1 ratio correlates with airflow obstruction in asthma and chronic bronchitis
Am J Respir Crit Care Med
(1998)
G. Mautino et al.
Increased release of matrix metalloproteinase-9 in bronchoalveolar lavage fluid and by alveolar macrophages of asthmatics
Am J Respir Cell Mol Biol
(1997)
Y.C. Lee et al.
The involvement of matrix metalloproteinase-9 in airway inflammation of patients with acute asthma
Clin Exp Allergy
(2001)
M. Bosse et al.
Serum matrix metalloproteinase-9: tissue inhibitor of metalloproteinase-1 ratio correlates with steroid responsiveness in moderate to severe asthma
Am J Respir Crit Care Med
(1999)

E.A. Kelly et al.

Increased matrix metalloproteinase-9 in the airway after allergen challenge

Am J Respir Crit Care Med

(2000)

B. Dahlen et al.

Immunohistochemical localisation of the matrix metalloproteinases MMP-3 and MMP-9 within the airways in asthma

Thorax

(1999)

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: Dr Broide is supported by National Institutes of Health grant AI 38425.

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Mechanisms of asthma and allergic inflammationRemodeling associated expression of matrix metalloproteinase 9 but not tissue inhibitor of metalloproteinase 1 in airway epithelium: Modulation by immunostimulatory DNA

Background

Objective

Methods

Results

Conclusion

Clinical implications

Section snippets

Induction of allergen-induced airway remodeling

Immunohistochemical detection of epithelial cell vs peribronchial cells expressing MMP-9 and TIMP-1 in the remodeled airway: Effect of ISS

Discussion

J Allergy Clin Immunol

J Allergy Clin Immunol

J Allergy Clin Immunol

Am J Pathol

Matrix metalloproteinases in lung biology

Respir Res

Contribution of 92 kDa gelatinase/type IV collagenase in bronchial inflammation during status asthmaticus

Am J Respir Crit Care Med

Sputum metalloproteinase-9/tissue inhibitor of metalloproteinase-1 ratio correlates with airflow obstruction in asthma and chronic bronchitis

Am J Respir Crit Care Med

Increased release of matrix metalloproteinase-9 in bronchoalveolar lavage fluid and by alveolar macrophages of asthmatics

Am J Respir Cell Mol Biol

The involvement of matrix metalloproteinase-9 in airway inflammation of patients with acute asthma

Clin Exp Allergy

Serum matrix metalloproteinase-9: tissue inhibitor of metalloproteinase-1 ratio correlates with steroid responsiveness in moderate to severe asthma

Am J Respir Crit Care Med

Increased matrix metalloproteinase-9 in the airway after allergen challenge

Am J Respir Crit Care Med

Immunohistochemical localisation of the matrix metalloproteinases MMP-3 and MMP-9 within the airways in asthma

Thorax

Mechanisms of asthma and allergic inflammation
Remodeling associated expression of matrix metalloproteinase 9 but not tissue inhibitor of metalloproteinase 1 in airway epithelium: Modulation by immunostimulatory DNA