Susumu Yasuoka

Localization of human airway trypsin-like protease in the airway: an immunohistochemical study

Abstract. Human airway trypsin-like protease (HAT) has been isolated from mucoid sputum of patients with chronic airway diseases. In order to clarify the cellular source of this novel protease in the human airway, we examined the localization of immunoreactive HAT in bronchial tissues obtained at surgery and fixed in 4% paraformaldehyde using an extremely sensitive immunohistochemical technique called a catalyzed signal amplification method and a monoclonal antibody against recombinant HAT. HAT immunoreactivity was demonstrated in cytoplasm of ciliated cells of bronchial epithelium and/or at the basal part of cilia. No positive reaction was found in submucosal glands or mast cells. The heterogeneous distribution of HAT immunoreactivity within the bronchial epithelium indicates that its expression might be changeable and that it might be closely related to the physiological status of the airway epithelium. Non-specific but intense reaction caused by endogenous avidin-binding activity (EABA) was selectively detected in submucosal glands, but was effectively blocked by successive treatments with avidin and biotin. These results indicate that HAT may be synthesized in the ciliated cells and that it may play some physiological roles within the epithelial layer and on the airway surface. It is necessary to keep in mind that some cells show strong EABA, especially when a highly sensitive immunohistochemical technique is applied.

Cytotoxic Factor for Fibroblasts in Bronchoalveolar Lavage Fluid of Bleomycin-Treated Rats

The mechanisms of lung injury and fibrosis in diffuse interstitial lung diseases were studied by tests on the in vitro effect of bronchoalveolar lavage fluid (BALF) from bleomycin-treated rats on quiescent fibroblasts in culture. The BALF was obtained on days 2, 3, 6, 15, and 29 after intratracheal administration of bleomycin. Cytotoxic activity was detected in the BALF only on day 2. The factor responsible for the cytotoxic activity was detected in 50-70% ammonium sulfate fraction, and lost 10% of its activity after 30 min at 56 degrees C, 75% after 10 min at 80 degrees C, and all activity after 10 min at 100 degrees C. The factor was completely precipitated by 60% acetone and was not extractable with ether. In ion exchange chromatography, the factor was eluted with the main fraction of protein, and gel filtration indicated that it had a molecular weight of 60-70 kDa. Histological examination showed disappearance of the normal alveolar architecture on day 2, and pulmonary fibrosis on day 15. Thus the cytotoxic factor may be one of factors responsible for lung injury and may act as trigger for subsequent fibrosis in bleomycin-induced lung damage.

Triglyceride metabolism in the lung.

Experiments were carried out to examine whether the lung acts as a depot for circulating lipid, especially that absorbed from the intestine. When 0.5 ml of triolein was administered orally to rats, the triglyceride content of the lung increased 2-3 hr later, but its increase in the lungs 2-3 hr later was only of about 1/10 of that in the liver. In the fed state the triglyceride content of the lung was only about 1/8 of that of the liver. When [3H]palmitic acid was administered orally to mice its uptake by the lung 1 and 2 hr later was 1/25-40 of that by the liver. In the lung, it was incorporated into phospholipid more than into triglyceride, but in the liver it was predominatly incorporated into triglyceride. Most of the lipase activity in both the microsomal and soluble fractions of rat lung appeared to be due to lipoprotein lipase. Fasting did not decrease the lipoprotein lipase activity in either fraction. It was concluded that the lung is not important in removal of triglyceride from the blood, even during fat absorption from the intestine, and that the lung takes up circulating lipid for its own metabolism rather than for storage.