Characterization of extended co-culture of non-typeable Haemophilus influenzae with primary human respiratory tissues

Author:

Ren Dabin1,Nelson Kevin L2,Uchakin Peter N1,Smith Arnold L2,Gu Xin-Xing3,Daines Dayle A1

Affiliation:

1. Division of Basic Medical Sciences, Mercer University School of Medicine, 1550 College Street, Macon, GA 31207

2. Seattle Children's Research Institute, 1900 Ninth Avenue, C9S-8, Seattle, WA 98101

3. National Institute on Deafness and Other Communication Disorders, 5 Research Court, Rockville, MD 20850, USA

Abstract

Non-typeable Haemophilus influenzae (NTHi) are human-adapted Gram-negative bacteria that comprise part of the normal flora of the human upper airway, but are also responsible for a number of mucosal infections such as otitis media and bronchitis. These infections often recur and can become chronic. To characterize the effect of long-term co-culture of NTHi with human tissues, we infected primary respiratory epithelial cells grown at the air–liquid interface with three NTHi strains over a range of 1–10 days. Scanning and transmission electron microscopy of tissues confirmed that intact NTHi were persisting paracellularly, while organisms observed in intracellular vacuoles appeared degraded. Furthermore, the apical surface and tight junctions of the infected tissues were undisturbed, with high transepithelial electrical resistances, while the basal cell layer displayed more junctional disorganization and wider intercellular spaces than the uninfected control tissues. Although the tissues elaborated the cytokine profile reported for NTHi-caused otitis media in vivo, there was little change in the dynamics of cytokine secretion over the time points tested. Finally, we report that NTHi strains released outer membrane vesicles (OMVs) during extended co-culture with the tissues, and show that these OMVs directly interact with host cell membranes.

Publisher

SAGE Publications

Subject

General Biochemistry, Genetics and Molecular Biology

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