Primary in vitro culture of porcine tracheal epithelial cells in an air-liquid interface as a model to study airway epithelium and Aspergillus fumigatus interactions.
Since the airway epithelium is the first tissue encountered by airborne fungal spores, specific models are needed to study this interaction. We developed such a model using primary porcine tracheal epithelial cells (PTEC) as a possible alternative to the use of primary human cells. PTEC were obtained from pigs and were cultivated in an air-liquid interface. Fluorescent brightener was employed to quantify the internalization of Aspergillus fumigatus conidia. Potential differences (Vt) and transepithelial resistances (Rt) after challenge with the mycotoxin, verruculogen, were studied. Primers for porcine inflammatory mediator genes IL-8, TNF-alpha, and GM-CSF were designed for a quantitative real-time PCR procedure to study cellular responses to challenges with A. fumigatus conidia. TEM showed the differentiation of ciliated cells and the PTEC ability to internalize conidia. The internalization rate was 21.9 ± 1.4% after 8 h of incubation. Verruculogen (10(-6) M) significantly increased Vt without having an effect on the Rt. Exposure of PTEC to live A. fumigatus conidia for 24 h induced a 10- to 40-fold increase in the mRNA levels of inflammatory mediator genes. PTEC behave similarly to human cells and are therefore a suitable alternative to human cells for studying interaction between airway epithelium and A. fumigatus.
Khoufache
Khaled
K
UPEC, UMR BIPAR, ENVA, AFSSA, Créteil, France.
Cabaret
Odile
O
Farrugia
Cécile
C
Rivollet
Danièle
D
Alliot
Annie
A
Allaire
Eric
E
Cordonnier
Catherine
C
Bretagne
Stéphane
S
Botterel
Françoise
F
D000818:Animals; D001232:Aspergillus fumigatus / Q000472:pathogenicity*; D018929:Cell Culture Techniques; D002478:Cells, Cultured; D004847:Epithelial Cells / Q000382:microbiology*; D005260:Female; D020869:Gene Expression Profiling; D054884:Host-Pathogen Interactions; D018836:Inflammation Mediators / Q000378:metabolism; D020545:Respiratory Mucosa / Q000166:cytology / Q000382:microbiology; D013552:Swine; D037521:Virulence Factors / Q000378:metabolism
D018836:Inflammation Mediators; D037521:Virulence Factors