Yasuhito Honda

Aberrant Appearance of Lung Surfactant Protein A in Sera of Patients with Idiopathic Pulmonary Fibrosis and Its Clinical Significance

Pulmonary surfactant protein A (SP-A) is known to be a major phospholipid-associated glycoprotein in pulmonary surfactant, which is specific to the lung. In this study, the SP-A concentrations in sera of patients with various lung diseases were determined using an enzyme-linked immunosorbent assay. Patients with idiopathic pulmonary fibrosis (IPF) and pulmonary alveolar proteinosis (PAP) exhibited prominently high concentrations of serum SP-A compared to those of other lung diseases and healthy volunteers, although there were significant increases in serum SP-A concentrations in patients with pulmonary tuberculosis, chronic pulmonary emphysema, diffuse panbronchiolitis and bacterial pneumonia compared to those of healthy volunteers. Successive measurement in 2 patients with IPF showed that serum SP-A levels reflect the disease activity of IPF. In patients with IPF, serum SP-A concentrations were significantly correlated with those of serum lactate dehydrogenase, whereas there were no significant correlations of serum SP-A concentrations with erythrocyte sedimentation rate, arterial oxygen saturation, vital capacity and carbon monoxide diffusing capacity. Determination of serum SP-A will contribute to diagnosing IPF and PAP, and may reflect the disease activity of IPF.

Changes in phospholipids in bronchoalveolar lavage fluid of patients with interstitial lung diseases.

We analyzed phospholipids of human bronchoalveolar lavage (BAL) fluids from patients with interstitial lung diseases; idiopathic pulmonary fibrosis (IPF), sarcoidosis, and eosinophilic granuloma (EG) and compared them to those of normal subjects. The content of phospholipid/ml of BAL fluid was significantly decreased in IPF. There was a significant decrease in phosphatidylglycerol (PG) and an increase in phosphatidylinositol (PI) in IPF but not in sarcoidosis and EG. Thus, the PG to PI ratio was significantly decreased in IPF. The dipalmitoyl species of phosphatidylcholine (PC) was found to be significantly decreased in IPF and sarcoidosis by molecular species analysis using high performance liquid chromatography. In contrast, the unsaturated species were increased in these diseases. The decrease in dipalmitoyl PC appeared to be a common feature in interstitial lung diseases.The changes in phospholipids in BAL fluids, especially decreases in DPPC and PG to PI ratio in IPF, appear to indicate that damage of alveolar Type II cells and/or of metabolic disturbance in pulmonary surfactant occurs in IPF.

Increased carcinoembryonic antigen concentrations in sera and bronchoalveolar lavage fluids of patients with pulmonary alveolar proteinosis.

Pulmonary alveolar proteinosis (PAP) is a rare disease in which alveoli are filled with lipoproteinaceous materials. We measured carcinoembryonic antigen (CEA) concentrations in bronchoalveolar lavage (BAL) fluids and sera from patients with PAP and from healthy volunteers (HV). Significantly increased CEA values were found in BAL fluids and sera from patients with PAP compared with those from HV. BAL fluid CEA values significantly correlated to serum CEA values in patients with PAP. Serum CEA values significantly correlated to serum lactate dehydrogenase activity and alveolar-arterial PO2 difference values in patients with PAP. Successive measurements of serum CEA showed that serum CEA values reflect the disease activity of PAP. The determination of serum CEA is useful for evaluating disease severity of PAP.

BAL Surfactant Protein A and Clara Cell 10-kDa Protein Levels in Healthy Subjects

Abstract. Lung surfactant protein A (SP-A) and Clara cell 10-kDa protein (CC10) are the most abundant proteins produced locally in the lower respiratory tract, as assessed in bronchoalveolar lavage (BAL) analysis. However, it is not known what factors influence SP-A and CC10 levels in BAL fluids, and the relationship between SP-A and CC10 levels in BAL fluids has been unclear. We measured SP-A and CC10 concentrations in BAL fluids from 11 healthy nonsmokers and 12 healthy smokers by enzyme-linked immunosorbent assays using specific antibodies. Mean SP-A and CC10 levels in BAL fluids of healthy smokers were significantly lower than those of healthy nonsmokers. SP-A values correlated significantly with CC10 and phospholipid values in BAL fluids. CC10 values tended to correlate with phospholipid values in BAL fluids. On BAL examinations using three 50-ml aliquots, the mean SP-A level in the second lavage was 2.0-fold and 2.4-fold, respectively, of that in the first and third lavages, and the mean CC10 level in the first lavage was 5.0-fold and 5.6-fold, respectively, of that in the second and third lavages. We conclude that BAL fluid SP-A and CC10 levels are influenced by the BAL methods and by cigarette smoking. There is a significant positive correlation between SP-A and CC10 values in BAL fluids of healthy subjects.

Alterations of acidic phospholipids in bronchoalveolar lavage fluids of patients with pulmonary alveolar proteinosis

We analyzed phospholipids, especially the acidic phospholipids, i.e. phosphatidylglycerol (PG) and phosphatidylinositol (PI), of bronchoalveolar lavage (BAL) fluids from patients with pulmonary alveolar proteinosis. A significant decrease in PG and a concomitant increase in PI were found in the phospholipid of the patients BAL fluids compared to that of normal subjects. Thus, the PG to PI ratio was significantly decreased in pulmonary alveolar proteinosis. These changes in the acidic phospholipids in BAL fluids of patients with pulmonary alveolar proteinosis appear to indicate that a switch-over in the synthesis of both acidic phospholipids from a same precursor CDP-diacylglycerol (CDP-DG) in alveolar Type II cells occurs in a such diseased state. However, since the molecular species profiles of PG and PI in the patients BAL fluids were distinctly different from each other, the PG and PI present in the alveoli of the patients appeared to be derived from different pools of CDP-DG in alveolar Type II cells.

Lung surfactant protein-A and carcinoembryonic antigen in pleural effusions due to lung adenocarcinoma and malignant mesothelioma.

Lung surfactant protein-A (SP-A) is a major phospholipid-associated glycoprotein in surfactant, and is a useful immunohistochemical marker for lung adenocarcinoma. Carcinoembryonic antigen (CEA) has not been immunohistochemically detected in mesothelioma. In pleural effusions due to malignant mesothelioma, very low concentrations of SP-A and CEA can be expected. We studied the value of combined determinations of CEA and SP-A in pleural fluid to distinguish between lung adenocarcinoma and mesothelioma. SP-A and CEA concentrations were measured in pleural effusions from 78 patients with lung adenocarcinoma and 10 with malignant mesothelioma. SP-A concentrations in pleural effusions due to lung adenocarcinoma and mesothelioma were 516 +/- 140 and 16.9 +/- 3.6 ng.ml-1 (mean +/- SEM), respectively. CEA concentrations in pleural effusions due to lung adenocarcinoma and mesothelioma were 239 +/- 92.4 and 1.7 +/- 0.3 ng.ml-1, respectively. SP-A values did not exceed 100 ng.ml-1 in any of 10 mesotheliomas, whilst in 37 of 78 lung adenocarcinomas they did. CEA values did not exceed 10 ng.ml-1 in any of 10 mesotheliomas, whilst in 53 of 78 lung adenocarcinomas they did. Increased values of SP-A and/or CEA were found in pleural effusions from 67 of 78 lung adenocarcinomas. It is concluded that a combination of CEA and SP-A assays in pleural effusions will be helpful for discriminating lung adenocarcinoma from mesothelioma.

Pulmonary surfactant protein A in pleural effusions.

Pulmonary surfactant protein A (SP‐A) is known to be a major phospholipid‐associated glycoprotein in pulmonary surfactant, which is specific to the lung. Immunohis‐tochemically, expression of SP‐A in tumor tissues is found in approximately 50% of patients with lung adeno‐carcinoma but not in the other histologic types of lung cancer or metastatic lung tumors. In this study, the SP‐A content of pleural effusions was determined using an enzyme‐linked immunosorbent assay. These results showed that approximately 40% of patients with lung ad‐enocarcinomas (27 of 67) had high levels of SP‐A (> 500 ng/ml) in their pleural effusions. By contrast, patients with other histologic types of lung cancers, adenocarci‐nomas of different primary sites, and tuberculosis had low levels of SP‐A in their pleural effusions. The determination of SP‐A in malignant effusions will contribute to distinguishing primary lung adenocarcinoma from ade‐nocarcinomas of miscellaneous origin.

Lipid Analysis and Surfactant-Associated Protein Expression in Lung Adenocarcinoma Cells from Pleural Effusion

Primary lung adenocarcinomas originate from the progenitor cells of peripheral airway cells. Alveolar type II cells and Clara cells are the major progenitor cells of peripheral airway cells. Alveolar type II cells produce a lipid-protein complex called surfactant, which contains surfactant proteins SP-A, SP-B, SP-C and SP-D. Phosphatidylcholine (PC) and phosphatidylglycerol (PG) are believed to be essential for the surfactant function. Clara cells also express SP-A, SP-B and SP-D but not SP-C. In this study we examined the properties of the cancer cells isolated from the pleural effusion of a patient with primary lung adenocarcinoma by analyzing lipids, proteins and mRNAs. The cancer cells, designated as LC117 cells, were isolated from the pleural effusion of a patient with primary lung adenocarcinoma. The percent distributions of [14C]-acetate incorporated into PC and PG in the cancer cells were 55.7 and 1.1%, respectively. The disaturated species in total PC was 46.2%. Immunoblotting analysis using anti-SP-D monoclonal antibody revealed that the pleural effusion from a patient with lung adenocarcinoma contained SP-D. We determined the concentrations of SP-A and SP-D by enzyme-linked immunosorbent assay. The pleural effusions from this patient and the media incubated with cancer cells exhibited significant levels of SP-D as well as SP-A. Reverse transcriptase-polymerase chain reaction demonstrated that the tumor cells expressed mRNAs for SP-C as well as the other surfactant proteins. The results demonstrate that tumor cells from lung adenocarcinoma express all of surfactant-associated proteins, indicating that LC117 cells originate from alveolar type II cells. This study indicates that the combination of analyses of lipids, proteins and mRNAs in the cancer cells isolated from pleural effusion is useful to understand the property of lung adenocarcinoma.

A case of bronchioloalveolar carcinoma ultrastructural and lipid‐biochemical studies

A case of bronchioloalveolar carcinoma was studied electron microscopically and lipid biochemically. Electron microscopic examination revealed that the tumor was composed of three different types of cells, undifferentiated cells, cells possessing lamellar inclusion bodies within the cytoplasm, and cells containing mucus droplets. The characteristic findings in the phospholipid profiles of this case of bronchioloalveolar carcinoma were a high content of saturated classes of phosphatidylcholine, especially of the dipalmitoyl type, and the occurrence of appreciable amounts of phosphatidylglycerol. These observations indicate that some of the tumor cells examined in this study had the characteristic feature of alveolar type II cell differentiation which is responsible for production of pulmonary surface‐active materials. The value of phospholipid analysis in assessing such tumors is emphasized.

Pulmonary lymphangitis carcinomatosa due to gastric adenosquamous carcinoma, with cytologic findings - A case report.

35歳の男性が乾性咳嗽と微熱を主訴とし, 当院を受診した. 理学所見では, 脈拍数が109/分と頻脈で, 胸部聴診上全肺にわたり気管支呼吸音の増強が認められた. 表在リンパ節は触知しなかった. 血液検査では肝機能の軽度障害, CRP (2+), CEA (EIA) 42.0ng/mlなどの異常がみられた. 胸部X線写真では両側肺門部から肺野にかけて小粒状陰影および浸潤陰影, 心陰影の拡大, 両側胸水の貯留がみられた. 気管支鏡検査では気管分岐部から右気管支全体にわたる発赤, 浮腫状変化, 小隆起性病変などが観察された. 喀痰細胞診では孤立散在性にオレンジGやライトグリーン好性で, 胞体が厚く粘液様空胞をもつ悪性細胞がみられ, 擦過細胞診では層状の配列を示す悪性細胞がみられた. また, PAS陽性物質が認められた. 剖検により, 本症例を胃原発の腺扁平上皮癌, 癌性リンパ管症, 癌性胸膜炎, 癌性心膜炎と診断した. この症例にみられた細胞診所見にっき検討を加えた.