Injurious effect of the eosinophil peroxide-hydrogen peroxide-halide system and major basic protein on human nasal epithelium in vitro.
Tissue injury is observed in allergic and nonallergic eosinophilic rhinitis, but the mechanism of this injury is unclear. Because eosinophils are prominent in biopsy specimens in these conditions, we hypothesized that they may participate in the injury process. Initially, we developed techniques to isolate and purify human nasal epithelial cells from turbinate biopsies to use as target cells for eosinophil granule products. Primary cultures from explants were characterized by electron microscopy and indirect immunofluorescence with a panel of primary monoclonal and polyclonal antibodies. These studies revealed the homogeneity of the cells and confirmed their epithelial nature. Cultured nasal epithelial cells were then exposed to either purified human eosinophil peroxidase, bromide, and glucose plus glucose oxidase, as a continuous source of hydrogen peroxide, or eosinophil major basic protein. Neither eosinophil peroxidase alone nor glucose plus glucose oxidase in the absence of eosinophil peroxidase were injurious, but the combined addition of eosinophil peroxidase, glucose/glucose oxidase, and bromide produced marked target cell lysis. This effect was time- and eosinophil peroxidase dose-dependent. Catalase and azide significantly inhibited the lysis of these cells, suggesting the eosinophil peroxidase-catalyzed products of halide oxidation mediated this form of injury. The addition of purified human eosinophil major basic protein also caused dose- and time-dependent lysis of the nasal epithelial cells but required longer incubation periods to effect injury. We hypothesize that the eosinophil peroxidase-hydrogen peroxide-halide system and major basic protein may injure the nasal epithelium in inflammatory conditions such as allergic and nonallergic eosinophilic rhinitis.
Ayars
G H
GH
Department of Medicine, University of Washington, Seattle 98195.
Altman
L C
LC
McManus
M M
MM
Agosti
J M
JM
Baker
C
C
Luchtel
D L
DL
Loegering
D A
DA
Gleich
G J
GJ
D001798:Blood Proteins / Q000633:toxicity*; D001965:Bromides / Q000633:toxicity*; D002478:Cells, Cultured; D047091:Eosinophil Granule Proteins; D047209:Eosinophil Peroxidase; D004804:Eosinophils / Q000201:enzymology; D004847:Epithelial Cells; D005947:Glucose / Q000633:toxicity; D005949:Glucose Oxidase / Q000633:toxicity; D006801:Humans; D006861:Hydrogen Peroxide / Q000633:toxicity*; D066298:In Vitro Techniques; D009297:Nasal Mucosa / Q000187:drug effects*; D010544:Peroxidases / Q000633:toxicity*; D012220:Rhinitis / Q000209:etiology; D012221:Rhinitis, Allergic, Perennial / Q000209:etiology; D012260:Ribonucleases
D001798:Blood Proteins; D001965:Bromides; D047091:Eosinophil Granule Proteins; D006861:Hydrogen Peroxide; D005949:Glucose Oxidase; D047209:Eosinophil Peroxidase; D010544:Peroxidases; D012260:Ribonucleases; D005947:Glucose