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Rhinovirus Disrupts the Barrier Function of Polarized Airway Epithelial Cells

¹ Department of Pediatrics and Communicable Diseases, ² Department of Molecular and Integrative Physiology, University of Michigan, Ann Arbor, Michigan; ³ California Pacific Medical Center Research Institute, San Francisco, California; ⁴ Department of Laboratory Medicine, University of California, San Francisco, California; ⁵ Department of Medicine, University of Vermont, Burlington, Vermont

Correspondence and request for reprints should be addressed to Marc B. Hershenson, M.D., University of Michigan, 1150 W. Medical Center Dr., Room 3570, MSRBII, Box 5688, Ann Arbor, MI 48109-0688. E-mail: mhershen{at}umich.edu

Rationale: Secondary bacterial infection following rhinovirus (RV) infection has been recognized in chronic obstructive pulmonary disease.

Objectives: We sought to understand mechanisms by which RV infection facilitates secondary bacterial infection.

Methods: Primary human airway epithelial cells grown at air–liquid interface and human bronchial epithelial (16HBE14o-) cells grown as polarized monolayers were infected apically with RV. Transmigration of bacteria (nontypeable Haemophilus influenzae and others) was assessed by colony counting and transmission electron microscopy. Transepithelial resistance (R_T) was measured by using a voltmeter. The distribution of zona occludins (ZO)-1 was determined by immunohistochemistry and immunoblotting.

Measurements and Main Results: Epithelial cells infected with RV showed 2-log more bound bacteria than sham-infected cultures, and bacteria were recovered from the basolateral media of RV- but not sham-infected cells. Infection of polarized airway epithelial cell cultures with RV for 24 hours caused a significant decrease in R_T without causing cell death or apoptosis. Ultraviolet-treated RV did not decrease R_T, suggesting a requirement for viral replication. Reduced R_T was associated with increased paracellular permeability, as determined by flux of fluorescein isothiocyanate (FITC)-inulin. Neutralizing antibodies to tumor necrosis factor (TNF)-, IFN- and IL-1β reversed corresponding cytokine-induced reductions in R_T but not that induced by RV, indicating that the RV effect is independent of these proinflammatory cytokines. Confocal microscopy and immunoblotting revealed the loss of ZO-1 from tight junction complexes in RV-infected cells. Intranasal inoculation of mice with RV1B also caused the loss of ZO-1 from the bronchial epithelium tight junctions in vivo.

Conclusions: RV facilitates binding, translocation, and persistence of bacteria by disrupting airway epithelial barrier function.

Key Words: COPD • exacerbation • Haemophilus influenzae • tight junction • ZO-1

AT A GLANCE COMMENTARY Scientific Knowledge on the Subject The mechanisms by which rhinovirus infection leads to secondary bacterial infection in patients with underlying respiratory diseases are not well understood. What This Study Adds to the Field Rhinovirus infection facilitates transmigration of bacteria across polarized airway epithelia by dissociating zona occludens-1 from the tight junction complex, thereby impairing barrier function of the epithelium.