No abstract

Immunofluorescence studies of type II alveolar epithelial cells indicate that they first appear in the pseudoglandular period of mouse lung development (around day 14.2). They are the only cell type to line the prospective pulmonary acinus at this time. The ultrastructural characteristics of this cell are defined by investigating embryos aged 13-16 days with transmission and scanning electron microscopy. Early embryonic type II cells appear as low-columnar or cuboid cells having large, approximately round nuclei and distinct ultrastructural features, including a well-developed Golgi apparatus with many associated vesicles, multivesicular bodies, dense bodies, and large apical and basal glycogen fields. These fields represent a distinctive property of the cell. Frequently, they show compartmentalization due to the presence of membrane systems, and association with dense bodies of various sizes.

The present study was performed to find out whether the type II alveolar epithelial cell or its precursor (an approximately cuboidal cell lacking multilamellar bodies) is present before the twentieth week of human gestation. For this purpose we used an antibody, SALS-Hu(E), which recognizes the human type II cell on the basis of surfactant-associated proteins. Application of SALS-HuE (by indirect immunofluorescence) to acetone-fixed frozen sections of fetal lung tissue gave a distinct staining of the cuboidal or low columnar epithelial cells lining the end-pieces of the tubular system of fetal lung (initially only a few): this staining started around weeks 10 to 12 after conception. Around week 16 some of the labeled epithelial cells appeared to be rather flat and by week 19 a combined cellular and linear fluorescence pattern was seen. Columnar epithelial cells of the prospective bronchial portion did not show this specific staining. Our results indicate that the type II cell or its precursor cell is indeed present in the pseudoglandular period of human lung development, i.e., starting around the tenth to twelfth week. This cell type lines the acinar tubule, the basic structure of the pulmonary acinus. Transformation of this cell type into the type I alveolar epithelial cell seems to start in week 16.

The epithelial cell types present in respiratory (= distal alveolarized) and terminal (= distal nonalveolarized) bronchioles in adult human lung were characterized with scanning and transmission electron microscopy (SEM, TEM) and light microscopic cytochemistry, using specific antibodies against surfactant protein SP-A and mucins, and Alcian blue/periodic acid-Schiff (AB/PAS) staining. In the respiratory bronchiole, two epithelial cell populations share the same basal lamina: one pseudostratified columnar with ciliated, secretory, and basal cells and the other predominantly simple cuboid with some interspersed flat (type I) cells. The columnar secretory cells show the ultrastructure of mucous cells. Light microscopically, they react with mucin antibodies and contain primarily periodate-reactive acid mucins. The mucous cells are the distal secretory cells described by Clara (1937). The cuboid cells are identified as type II (precursor) cells based on ultrastructural criteria for embryonic type II cells (Ten Have-Opbroek et al., 1988a, 1990a), including a cuboid cell shape, a large and roundish nucleus, rough and smooth endoplasmic reticulum (ER), osmiophilic multivesicular bodies, and dense bodies. These dense bodies in turn frequently exhibit--like those in embryonic type II cells--internal vesicles or lamellae, variability in size and shape, a specific relationship to ER and a widespread cytoplasmic distribution. Finally, the cuboid cells show a cytoplasmic staining pattern for SP-A. The terminal bronchiole is lined by the columnar cell population. In the respiratory bronchiole, the columnar (bronchial) and cuboid (alveolar) cell populations occupy distinctly different zones (pulmonary artery zone versus remaining wall). The alveolar part of the respiratory bronchiole (called alveolar tubule) defines the proximal border of a true respiratory unit.

An indirect immunofluorescence technique was applied to frozen sections of central and peripheral human lung tissue to search for extracellular localizations of antileukoprotease (ALP). Two monoclonal anti-ALP antibodies recognizing different epitopes and polyclonal anti-ALP antibodies were used. ALP was found to be localized along elastic fibers in alveolar septa, and also along elastic fibers in the walls of bronchi, bronchioles and blood vessels. Serous cells of bronchial submucosal glands showed labelling as well. In frozen sections of liver and spleen no label was found. Cells and elastic fibers were not labelled when lung tissue sections were processed with polyclonal or monoclonal anti-ALP antibodies, that were blocked with purified ALP before the immunostaining. The association of ALP with elastic fibers of human pulmonary connective tissue is of importance in understanding the role of the inhibitor in the defense of the lung parenchyma against the action of proteolytic enzymes, which is thought to result in emphysema.