Use of mucin antibodies and cDNA probes to quantify hypersecretion in vivo in human airways.
Mucus hypersecretion is a prominent feature of the airway's response to injury. The ability to quantitatively detect mucin and mucin mRNA in vivo in human airways would facilitate the determination of safe exposure levels to various air pollutants and the identification of drugs capable of attenuating mucus hypersecretion. To this end, we have developed two assays: an enzyme-linked immunosorbent assay (ELISA) quantifying mucin-like molecules and a polymerase chain reaction (PCR)-based assay quantifying mucin mRNA. These tests are performed on bronchial lavage fluid and epithelial cells brushed from the surfaces of human airways at bronchoscopy. The PCR data are normalized to eliminate potentially confounding effects of nonepithelial cells in the samples. In a study of six smokers and six nonsmokers, the ELISA detected significantly more mucin-like material in the airways of the smokers than of the nonsmokers. The median mucin concentration for the smokers was 52.2 micrograms/ml (range, 16.3 to 4,860.0), whereas that for the nonsmokers was 12.7 micrograms/ml (range, 4.5 to 22.9). The difference between smokers and nonsmokers was statistically significant (P < or = 0.01). The PCR-based test showed a trend for RNA samples from smokers to be enriched (vis-à-vis nonsmokers) in mucin mRNA.
Steiger
D
D
Department of Anatomy, University of California, San Francisco 94143.
Fahy
J
J
Boushey
H
H
Finkbeiner
W E
WE
Basbaum
C
C
D000328:Adult; D000368:Aged; D000911:Antibodies, Monoclonal; D001483:Base Sequence; D015139:Blotting, Southern; D001980:Bronchi / Q000378:metabolism*; D001992:Bronchoalveolar Lavage Fluid / Q000276:immunology; D015342:DNA Probes; D018076:DNA, Complementary; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D009077:Mucins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D016133:Polymerase Chain Reaction / Q000379:methods; D015203:Reproducibility of Results; D012907:Smoking / Q000276:immunology
D000911:Antibodies, Monoclonal; D015342:DNA Probes; D018076:DNA, Complementary; D009077:Mucins