Tissue factor procoagulant expression by rat alveolar epithelial cells.

Fibrin deposition in the alveolar space is characteristic of inflammatory lung injury. The formation of fibrin in the alveolus results from the coagulation of extravasated plasma. The cellular elements that promote intra-alveolar clotting have not been completely defined. We have investigated the capacity of alveolar epithelial cells (AEC) to promote coagulation through the expression of procoagulant activity (PCA) in tissue culture. Using a single-stage coagulation assay, rat AEC monolayers were found to contain 20,750 +/- 4,035 procoagulant units (PCU)/10(6) cells; 10- to 20-fold greater activity than that found in concomitantly isolated alveolar macrophages. The epithelial-derived procoagulant was shown to be tissue factor by a series of assays using clotting factor-deficient human plasmas. Freshly isolated AEC also possessed PCA (2,500 +/- 1,000 PCU/10(6) cells) and expressed a 2.1-kb mRNA that hybridized with a cDNA for murine tissue factor. Using a kinetic turbidometric assay of clot acceleration, PCA was found on the surface of unstimulated epithelial monolayers and could be increased to 170% of control by incubation with phorbol myristate acetate (PMA). This response to PMA was accompanied by a parallel increase in the relative abundance of tissue factor mRNA. AEC shed particulate PCA into the culture media that displayed a specific activity similar to that recovered from alveolar lining fluid. Therefore, by expressing both cell surface and particulate PCA, the alveolar epithelium likely contributes significantly to the modulation of intra-alveolar coagulation.     

Distribution of proSAAS-derived peptides in rat neuroendocrine tissues.

Using a technique to identify substrates of the peptide processing enzyme carboxypeptidase E (CPE), several novel peptides were detected in the brain and pituitary of Cpe(fat)/Cpe(fat) mice and found to be derived from a single precursor, termed proSAAS. In order to gain further information regarding the potential physiological roles of these peptides, we have examined the distribution of two proSAAS-derived peptides, ARPVKEPRSLSAASAPLAETSTPLRL (SAAS) and LENSSPQAPARRLLPP (LEN), in rat neuroendocrine tissues using immunohistochemistry. Both peptides are detected throughout the brain, with the highest concentrations of SAAS peptide in the hypothalamus. In the hippocampus, both peptides are co-localized with prohormone convertase 1 in the dentate gyrus and CA1-3 region. In cerebellum, SAAS peptide is co-localized with prohormone convertase 1 in Purkinje and granular cells, whereas LEN is much more abundant in the Purkinje cells relative to the granular cells. Similarly, SAAS and prohormone convertase 1 are co-localized in the dorsal horn of the spinal cord, while LEN is mainly restricted to fibers of the white matter. In the pituitary, SAAS, LEN, and prohormone convertase 1 are detected in all three lobes. In the pancreas, SAAS, LEN, and prohormone convertase 1 are only detected in the islets, although the peptides are enriched in the peripheral cells (alpha and/or delta) while prohormone convertase 1 is only expressed in the inner cells (beta). Both SAAS and LEN are present in the adrenal medulla along with prohormone convertase 1. Taken together, these data are consistent with the proposed role for proSAAS as an endogenous inhibitor of prohormone convertase 1 in many, but not all cell types. However, the broader localization of the peptides allows for the possibility that they perform additional functions.     

Calcitonin gene-related peptide-immunoreactive nerves and neuroendocrine cells after lung transplantation in the rat.

Bronchial innervation is interrupted at lung transplantation. Nerve fibers with cell bodies above the section, such as sensory C fibers, should degenerate. Using histofluorescence, we evaluated calcitonin gene-related peptide (CGRP) immunoreactivity in syngeneic Lewis rats 1 and 5 mo after unilateral lung transplantation and in controls. CGRP-immunoreactive (IR) neuroendocrine cells were located within the epithelium of large and small bronchi. At 1 mo after transplantation, their number had significantly increased in large bronchi and had normalized 5 mo after transplantation. The density of CGRP-IR fibers in control lungs gradually decreased from large (0. 35 +/- 0.02 micron/micron basal lamina) to small (0.23 +/- 0.02) and peripheral bronchi (0.12 +/- 0.01). At 1 mo after lung transplantation, few CGRP-IR fibers were observed in large bronchi (0.17 +/- 0.02), fewer in small bronchi (0.04 +/- 0.01) (P < 0.01), and none in peripheral bronchi. At 5 mo after lung transplantation, transplanted lungs still had fewer CGRP-IR fibers in large (0.22 +/- 0.02) and small (0.11 +/- 0.02) bronchi (P < 0.02) than did controls, but there were, nonetheless, more in the small bronchi than at 1 mo after transplantation (P < 0.01). Additionally, few CGRP fibers were present in the peripheral bronchi (0.03 +/- 0.01) (P < 0.01). These results clearly demonstrate the occurrence of denervation followed by partial reinnervation with CGRP-IR fibers after transplantation in rat lung.     

Alterations in eicosanoid production by rat alveolar type II cells isolated after silica-induced lung injury.

Although alveolar type II cells in primary culture have been shown to produce eicosanoids and exposure of type II cells to silica in vitro alters eicosanoid production, the production of eicosanoids by alveolar type II cells isolated after acute lung injury in vivo has not been evaluated. Therefore, we investigated the production of arachidonic acid (AA) metabolites by alveolar type II cells isolated after silica-induced lung injury. Alveolar type II cells were isolated from rats 14 days after intratracheal silica instillation and from untreated animals. Type II cells were separated into normotrophic and hypertrophic populations by centrifugal elutriation, and secreted eicosanoids were determined under basal and stimulated conditions by enzyme immunoassay on the day of isolation and after 1 day in culture. Under basal conditions, freshly isolated type II cells from silica-treated animals produced more prostaglandin (PG) E2 than 6-keto-PGF1 alpha or thromboxane B2 (TxB2). Production of all three prostanoids increased with increasing cell size. The calcium ionophore A23187 stimulated a less than 2-fold increase in PGE2 and 6-keto-PGF1 alpha production in all groups of cells. In contrast, this calcium ionophore greatly enhanced TxB2 and leukotriene C4 (LTC4) production by normotrophic type II cells from both untreated and silica-treated animals. Incubation with exogenous AA suggested that the increased capability of the hypertrophic cells to synthesize PGE2 and TxB2 was due primarily to an increase in arachidonate availability. The hypertrophic type II cells also appear to have increased prostacyclin synthase activity. There were no differences in the catabolism of PGE2 between the normotrophic and the hypertrophic type II cells.(ABSTRACT TRUNCATED AT 250 WORDS)     

Altered expression of adhesion molecules and epithelial-mesenchymal transition in silica-induced rat lung carcinogenesis

Loss of the epithelial phenotype and disruption of adhesion molecules is a hallmark in the epithelial-mesenchymal transition (EMT) reported in several types of cancer. Most of the studies about the relevance of adhesion and junction molecules in lung cancer have been performed using established tumors or in vitro models. The sequential molecular events leading to EMT during lung cancer progression are still not well understood. We have used a rat model for multistep lung carcinogenesis to study the status of adherens and tight junction proteins and mesenchymal markers during EMT. After silica-induced chronic inflammation, rats sequentially develop epithelial hyperplasia, preneoplastic lesions, and tumors such as adenocarcinomas and squamous cell carcinomas. In comparison with normal and hyperplastic bronchiolar epithelium and with hyperplastic alveolar type II cells, the expression levels of E-cadherin, alpha-catenin and beta-catenin were significantly reduced in adenomatoid preneoplastic lesions and in late tumors. The loss of E-cadherin in tumors was associated with its promoter hypermethylation. alpha- and beta-catenin dysregulation lead to cytoplasmic accumulation in some carcinomas. No nuclear beta-catenin localization was found at any stage of any preneoplastic or neoplastic lesion. Zonula occludens protein-1 was markedly decreased in 66% of adenocarcinomas and in 100% squamous cell carcinomas. The mesenchymal-associated proteins N-cadherin and vimentin were analyzed as markers for EMT. N-cadherin was de novo expressed in 32% of adenocarcinomas and 33% of squamous cell carcinomas. Vimentin-positive tumor cells were found in 35% of adenocarcinomas and 88% of squamous cell carcinomas. Mesenchymal markers were absent in precursor lesions, both hyperplastic and adenomatoid. The present results show that silica-induced rat lung carcinogenesis is a good model to study EMT in vivo, and also provide in vivo evidence suggesting that the changes in cell-cell adhesion molecules are an early event in lung carcinogenesis, while EMT occurs at a later stage.     

Polyfunctional role of the alveolar brush cells in the rat lung

An investigation by scanning and transmission electron microscopy revealed an increase in the number of vacuoles in the apical part of the cytoplasm in the alveolar brush cells of the regenerating rat lung, hyperplasia of the lamellar complex, and activation of the protein-synthesizing apparatus. Immature surfactant material and secretory granules with an osmiophilic center were found in the cytoplasm of the brush cells. Colchicine, injected intramuscularly into rats six times in the course of the 24 h before sacrifice in a dose of 0.1 mg/100 g body weight, affected neither the number, topography, nor the structure of the bundles of microfibrils present in large numbers in the brush cells. Meanwhile, under the influence of colchicine, some of the microvilli of the alveolar brush cells undergo destruction and resorption. These data on the ultrastructural organization of these cells indicate that they can perform several functions: absorptive, contractile, and secretory.Department of Geographic Pathology, Institute of Human Morphology, Academy of Medical Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR A. P. Avtsyn.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 88, No. 10, pp. 485–489, October, 1979.     

Bcl-2 protein expression in lung cancer and close correlation with neuroendocrine differentiation.

For determination of the cellular distribution of bcl-2 expression in lung cancer and clarification of its correlation with cell neuroendocrine differentiation, Bcl-2 immunostaining was carried out on a large series of formalin-fixed, paraffin-embedded lung cancer samples, and four general neuroendocrine marker and seven peptide hormone stainings were carried out on all Bcl-2-positive squamous cell carcinomas and adenocarcinomas of the lung as well as on 8 pulmonary neuroendocrine carcinomas histologically diagnosed. In addition, 3 small cell lung cancer cell lines were studied by Western blotting. Neuroendocrine differentiation in Bcl-2-negative squamous cell carcinomas and adenocarcinomas was examined with chromogranin A and alpha-subunit of Go protein stainings. Bcl-2 protein was detected in 104/111 small cell carcinomas, 8/8 neuroendocrine carcinomas, 0/6 typical (well differentiated) carcinoids, 23/64 squamous cell carcinomas, 4/65 adenocarcinomas, and all 3 small cell lung cancer cell lines. All 8 neuroendocrine carcinomas, 11 of the Bcl-2-positive squamous cell carcinomas, and all 4 Bcl-2 positive adenocarcinomas expressed multiple neuroendocrine markers. The distributions of Bcl-2 and neuroendocrine marker immunoreactivity closely paralleled each other on consecutive sections. In squamous cell carcinomas, Bcl-2-positive cells could be roughly subdivided into those with neuroendocrine differentiation features, usually demonstrating intense Bcl-2 staining, with basaloid tumor cells usually expressing weak to moderate Bcl-2 staining. The present study clearly shows Bcl-2 protein expression to be remarkably differentially regulated according to histological types of lung cancers and to appear to quite likely be closely associated with neuroendocrine differentiation of tumor cells, indicating that bcl-2 is importantly involved in cell development and differentiation, in addition to protecting cells from apoptosis. Bcl-2 might be usable as a neuroendocrine marker in lung cancers and possibly also in neural-crest-derived tumors.     

Oncodevelopmental expression of rat placental alkaline phosphatase. Detection in oval cells during liver carcinogenesis.

Oval cells isolated from livers of rats fed a choline-deficient diet containing 0.1% DL-ethionine (CDE) have an alkaline phosphatase (ALKP) isozyme which can be distinguished by its electrophoretic mobility from the enzyme present in parenchymal cells isolated from normal liver or livers of rats fed the CDE diet for 4 weeks. The oval cell ALKP has the same electrophoretic mobility as the enzyme from fetal rat liver and placenta. ALKPs from oval cells, parenchymal cells, and placenta all differ from the intestinal enzyme by their electrophoretic mobility, isoelectric focusing, and the patterns of amino acid inhibition of enzyme activity. Oval cells in preneoplastic livers, fetal hepatocytes, and tumor cells of a primary hepatocellular carcinoma induced by CDE feeding stained with a monoclonal antibody directed against rat placental ALKP. Hepatocytes (in normal or preneoplastic livers) and bile duct cells in normal liver did not stain with the same antibody. Placental ALKP may thus be a useful marker in tracing the origin and fate of oval cells during hepatocarcinogenesis.     

Light-optical and electron-microscopic study of alveolar brush cells of the rat lung

In semithin sections through rat lung tissue stained metachromatically with toluidine blue 52 alveolar brush cells (ABC) were discovered. During a light-optical study the distinguishing features of these cells were the pyramidal shape of their body, the basal position of the nucleus, the darker staining of the cytoplasm than in other alveolar cells, and the presence of microvilli on the small free surface of the cell. For every 21 type II and 15 type I alveolar cells there was one ABC. Of the total number of ABC 41.1% are located at junctions between neighboring alveolar cells, 32.7% on the alveolar wall facting the cavity of one alveolus, 16.8% near the entrance to the alveoli, and 9.4% facing the lumen of two adjacent alveoli simultaneously or near Cohn's pores. Parallel electron-microscopic investigations revealed a granular cytoplasmic reticulum in ABC of an unusual type for other alveolar cells; it consisted of blocks of 5 to 8 apparently confluent tubules, bundles of fibers microtubules, and vacuoles in the apical cytoplasm. The structural organization of the ABC, their topography, and the frequency with which they were found in the alveoli of rats are evidence that these cells are chemoreceptor in nature.Laboratory of Geographic Pathology of the Baikal-Amur Trunk Railroad Zone, Institute of Human Morphology, Academy of Medical Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR A. P. Avtsyn.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 86, No. 11, pp. 592–596, November, 1978.     

Diabetic pregnancy. Changes in lectin binding to the surface of rat lung alveolar epithelial cells.

The binding of the lectins concanavalin A (Con A) and wheat germ agglutinin (WGA) to the luminal surface of lung alveolar epithelial cells was compared in normal rats and rats with streptozotocin-induced diabetes and their offspring. Lung tissue was lavaged, then fixed in situ with 3% glutaraldehyde. Buffer-rinsed slices of lung were incubated in Con A, WGA, or various control media. Lectin binding sites were visualized by the use of the peroxidase method. Normal neonates and those that were the results of diabetic pregnancies showed a hexose-specific Con A and WGA binding pattern qualitatively similar to that of normal and diabetic adults, respectively. In the normal animals, Con A binding sites were masked by sialic acid residues and were removable with alpha-mannosidase after neuraminidase treatment. In the diabetic adults and their offspring, one the other hand, Con A binding sites were readily accessible and were totally removed only by sequential treatment with alpha-mannosidase and alpha-glucosidase. WGA binding was essentially eliminated with neuraminidase in all animals except in the neonates from diabetic pregnancies, where N-acetyl-glucosaminidase was also required. The effects of maternal diabetes were reversible and occurred about Day 7 postpartum in the neonate. The effects were also reversible following insulin replacement in the diabetic adult.     

Surfactant protein C is expressed in alveolar type II cells but not in Clara cells of rat lung.

The expression of the surfactant-associated proteins in bronchiolar cells remains to be defined. We used in situ hybridization to identify sites of message expression of the surfactant-associated proteins A, B, and C (SP-A, SP-B, and SP-C) in adult and fetal rat lung. The expression of these messages by in situ hybridization was also compared with the localization of SP-A by immunocytochemistry. The localization of SP-A was used to identify type II cells and nonciliated bronchiolar epithelial (Clara) cells in these sections. The cRNA antisense probes for SP-A, SP-B, and SP-C appeared to hybridize over type II cells. Sense probes showed no localization or apparent specific hybridization. Messages for both SP-A and SP-B were also found in nonciliated bronchiolar epithelial (Clara) cells. However, no message for SP-C was observed in these cells. Clara cells from terminal to large bronchioles lacked detectable mRNA for SP-C. Expression of surfactant protein mRNAs was not detectable in type I cells, alveolar macrophages, interstitial cells, or vascular cells. Similarly, in fetal rat lung the messages for SP-A and SP-B but not SP-C were detected in bronchiolar cells. We conclude that rat Clara cells do not express SP-C mRNA, and thus SP-C can be regarded as a specific marker for rat type II cells.     

腺病毒E1A蛋白对大鼠肺泡上皮细胞谷胱甘肽活性的影响及其机制探讨

目的 研究腺病毒E1A蛋白潜伏感染对大鼠肺泡上皮细胞谷胱甘肽(GSH)活性的影响及其机制.方法 构建稳定表达腺病毒E1A蛋白的大鼠肺泡上皮细胞,观察氧化剂刺激时细胞内GSH含量变化,检测GSH合成限速酶γ-谷氨酰半胱氨酸合成酶(γ-GCS)催化活性的变化,通过观察γ-GCS 催化亚单位(GCLC)基因蛋白表达水平、mRNA表达水平和5′-上游调控序列调控的荧光素酶活性的改变,了解影响酶活性改变的机制.结果 腺病毒E1A蛋白表达抑制氧化应激时GSH的合成,抑制γ-GCS 催化活性和蛋白表达水平,抑制GCLC 基因mRNA表达,与5′-上游调控序列报导系统获得的结果一致.结论 腺病毒潜伏感染可能通过抑制GCLC基因 5′-上游调控序列的促转录活性,抑制了γ-GCS的表达和催化活性,从而抑制了氧化应激时GSH的合成,削弱机体的抗氧化作用,加重氧化损伤,可能是腺病毒潜伏感染参与慢性阻塞性肺病(COPD)发病的机制之一.     

The freeze-fracture study of alveolar type II cells and alveolar content in fetal rabbit lung.

Freeze-fracture replication technique was utilized to study the morphology of type II alveolar epithelial cells and alveolar contents in the late gestation of rabbit fetuses. It was shown that the lamellar inclusion bodies of type II cells were enveloped by the usual type of unit membrane with membrane-associated particles of 15 nm diameter. The interior of the inclusion bodies was composed of multiple stacks and/or whorls of membranes which were devoid of membrane-associated particles. Small vesicles within the inclusion were found more frequently with this technique than in chemically fixed thin-sectioned preparations. The intra-alveolar contents were comprised of two components; sperical bodies, which were identical to the internal contents of the lamellar bodies of type II cells, and tubular elements. These tubules most often appeared rectangular on cross-fractured faces. Triangular hexagonal fracture faces were also noted. The tubules were seen to rest on the surfaces of the spherical components. Our observations suggest that the tubular element of alveolar contents is formed through the interaction between the discharged lamellar body content and the alveolar fluid, and further suggest that at least the major constituent of type II cell lamellar bodies is lipid not bound to protein. Three new observations were made in this study; the absence of membrane-associated particles on the interior of the lamellae of the inclusions, the cross-fractured profiles of tubular elements of the alveolar contents, and the occasional multicompartmental nature of type II cell inclusions.     

Abnormal p53 expression in lung neuroendocrine tumors. Diagnostic and prognostic implications.

The diagnostic and prognostic implications of p53 immunostaining have been investigated in 59 pulmonary neuroendocrine tumors, including typical carcinoids (n = 15), so-called "atypical carcinoids" (n = 22), and small cell lung carcinomas (SCLCs; n = 22). Immunocytochemistry was performed on formalin-fixed, paraffin-embedded samples using the monoclonal antibody PAb1801, which has been shown to be suitable for staining fixed and embedded tissue sections. p53 immunoreactivity was restricted to atypical carcinoids (45% of the cases being immunoreactive) and to SCLCs (which were positively stained in 59% of the cases), whereas it was consistently lacking in typical carcinoid tumors. When the group of the so-called "atypical carcinoids" was further reclassified, p53 immunostaining was strictly confined to those cases belonging to the histologically more aggressive subsets (well differentiated neuroendocrine carcinoma subsets II and III). Within the same tumor type, however, p53 immunoreactivity did not correlate with the clinical outcome of the disease and was not predictive of the length of survival. The data indicate that abnormal p53 expression (which is strictly dependent on structural abnormalities of the p53 gene) is detectable in the majority of neuroendocrine carcinomas of the lung and might represent a useful adjunct in the differential diagnosis of pulmonary neuroendocrine neoplasms, particularly in routinely fixed and embedded small bronchoscopic biopsies.     

Antigen expression of alveolar macrophages in smokers and patients with lung diseases

Alveolar macrophage function was studied immunocytochemically using three monoclonal antibodies—macrophage CD 68 KP 1 (M), protein CD 11C (P), and anti-elastin (EL)—and three polyclonal antibodies—lysozyme (LZ), alpha-1-antitrypsin (AAT), and alpha-1-antichymotrypsin (AACT). the material for study was smears obtained from bronchial washings from 15 healthy persons and 60 patients with respiratory infections or primary or secondary malignant lung infiltration. Eight of the healthy group and 40 of the patient group were smokers (SM). the percentage of cells obtained from the washings which were macrophages was also measured. The intensity of staining reactions for each of the six antigens was noted and in general more intense staining was noted in smokers than in non-smokers. More intense staining was observed in patients with pulmonary infections (group II PI) and metastatic pulmonary infiltrations (group IV MP Ca) than in controls (group IC), while patients with primary lung cancer (group III PP Ca) had highly reduced staining reactions. the number of macrophages was similarly increased in all groups in comparison with the IC group for non-smokers and in all groups except III PP Ca for smokers. It is concluded that smoking, pulmonary infections, and metastatic infiltration of the lung are associated with an increase in the number and activity of alveolar macrophages, while patients with primary lung cancer have an increase in the number of macrophages which are functionally incompetent.     

Secretion of lung fluid by the developing fetal rat alveolar epithelium in organ culture.

We studied lung explants in submersion organ culture to examine the role of the developing fetal alveolar epithelium in the production of lung fluid. Fourteen-day-gestation fetal rat lungs were grown in a collagen gel matrix supplemented with F-12 media and 10% fetal calf serum. In this model, the lung continues to grow, secrete fluid, and become progressively cystic in morphology. There is gradual thinning of the distal epithelial layer, which is lined by alveolar type II cells and their precursors. After 6 to 8 days in culture, we impaled the cyst walls with a microelectrode and continuously recorded the transepithelial potential (psi t). Stable, baseline transepithelial potentials of -1.1 to -6.2 mV (mean +/- SEM = -3.3 +/- 0.11 mV, lumen negative, n = 34) were measured in bicarbonate-buffered Ringer's solution, suggesting active electrolyte transport. When bumetanide, an inhibitor of chloride secretion in other systems, was added to the bathing solution, psi t decreased from a baseline of -3.5 +/- 0.07 mV (mean +/- SEM) to a value of -2.2 +/- 0.07 mV, suggesting chloride transport contributes to the voltage (n = 18, P less than 0.0005). Isoproterenol hyperpolarized psi t from a baseline of -4.3 +/- 1.0 mV to -6.5 +/- 1.0 mV (n = 7, P less than 0.005). 8-(4-Chlorophenylthio) adenosine 3':5'cyclic monophosphate (CPT-cAMP) plus isobutylmethylxanthine (IBMX) similarly hyperpolarized psi t from a baseline of -4.6 +/- 0.4 mV to -7.3 +/- 0.7 mV (n = 11, P less than 0.005). Addition of bumetanide after stimulation with isoproterenol or CPT-cAMP/IBMX depolarized psi t.(ABSTRACT TRUNCATED AT 250 WORDS)     

Improved isolation of rat lung alveolar type II cells. More representative recovery and retention of cell polarity.

We have developed an improved isolation procedure for rat lung alveolar Type II (ATII) cells which yields a more representative sample of ATII cells with respect to their densities. This procedure includes an initial selection on a density gradient of approximately the complete density range of rat lung ATII cells. Subsequently, to exclude contaminating macrophages and lymphocytes from this fraction, the authors have exploited the fact that the contaminating cells have leukocyte common antigen (LC) on their surfaces, whereas the ATII cells do not. Our procedure yields 2 X 10(7) ATII cells per rat in a fraction which is 90% pure; the cells are immediately available for biochemical or pharmacologic analysis and represent a 90-95% recovery of the ATII cells loaded onto the density gradient. The cells retain their in vivo morphologic characteristics, including their polarity.     

Origin of ciliated alveolar epithelial cells in bleomycin-induced lung injury.

Bleomycin is known to induce diffuse pulmonary fibrosis and epithelial metaplasia. The reaction of the alveolar epithelium following a single intravenous or multiple intraperitoneal injections of bleomycin to mice is now examined in a combined morphologic and cytodynamic study. Necrosis of Type 1 cells was observed, followed by proliferation of Type 2 cells, a common reparative process. The proliferated cells transformed to a variety of epithelial forms, including ciliated cells and cells with morphologic features intermediate between alveolar and bronchiolar epithelium. No evidence of cell injury or increased cell division was found in the bronchial epithelium. It is concluded that the metaplastic ciliated epithelial cells are produced by an abnormal reparative process in the alveolar epithelium. The results suggest that, whereas the "resting" Type 2 cell is not vulnerable to bleomycin, in the postmitotic phase the drug may modify the synthetic mechanisms of cellular differentiation and thereby induce metaplasia.