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Pulmonary surfactant consists of phospholipids and four surfactant proteins, designated as SP-A, -B, -C, -D. SP-A is a major surfactant associated glycoprotein which is specific to the lung. In this study, we measured SP-A in sera and in bronchoalveolar lavage （BAL） fluid in patients with idiopathic interstitial pneumonia （IIP）, using ELISA, and compared those to various clinical data. Moreover, we investigated the local state of SP-A in HP lesions, using lung biopsied materials. Serum SP-A levels correlated with serum lactate dehydrogenase （LDH） activity, and serum SP-A in patients with accelerated erythrocyte sedimentation rate （ESR） was significantly higher than that in patients without accelerated ESR. Serum SP-A in patients with more than three years survival was also significantly higher than that in patients who died in less than three years, suggesting that serum SP-A value reflects the disease activity of IIP. Immunohistochemical studies showed that SP-A was not detected in bronchioles where alveoli had collapsed, and many hyperplastic type II cells strongly expressed SP-A in alveolar ducts and this was thought to be associated with high levels of serum SP-A. However, the quantity of SP-A in tissue homogenates significantly decreased in IIP compared to that in normal lung, and SP-A in BAL fluid negatively correlated to those in sera. Western blot analyses showed that there were no abnormal SP-As compared to normal control either in lung homogenates or in BAL fluids. We concluded that serum SP-A concentrations increase in accordance with the activity of IIP since alveolar to vascular leakage was accelerated in IIP.

Current cocepts of the pathogenesis of fibrotic changes in pulmonary asbestosis implies the roles of alveolar macrophages and cytokines produced by themselves. Tumor necrosis factor α(TNFα) and interleukin-1 β(IL-1β) are multifunctional cytokines produced by alveolar macrophages (AMs), cytokines which regulate inflammation and fibrosis. The aim of this study was to evaluate the role of TNFα and IL-1β secreted by AMs in the pathogenesis of pulmonary asbestosis. We isolated AMs by bronchoalveolar lavage from 11 subjects exposed to asbestos (AE) for a long time (32±8yrs) and from 7healthy controls (HC). AMs were stimulated with/without lipopolysaccharide (10μg/ml) for 24h and supernatants were assayed for TNFα and IL-1β by enzyme linked immunosorbent assay. AMs from AE produced higher levels of TNFα and 1L-1β upon stimulation with lipopolysaccharide than did the cells of HC. TNFα concentrations in BALFs were significantly higher in AE than in HC. IL-1β concentrations in BALFs were very low, and there was no significant difference between AE and HC. TNFα concentrations in BALFs and TNFα production from AMs positively correlated with the percentage of neutrophil in BALFs. AE were divided into two groups by roentogenographic profuound according to the ILO/UC classification (progressive group : more than 2, early group : less than Ⅰ). TNFα concentrations in BALFs were significantly higher in the progressive group than in the early group. In conclusion, these results indicate that TNFα is constitutively secreted by AMs from AE and is possibly associated with the pathogenesis of alveolitis and fibrosis due to asbestos exposure.

To elucidate the cause of severe asthma in adults, an animal model of chronic asthma was established in guinea pig sensitized by intraperitoneal injection of Ascaris suum and inhalation of the antigen repeated ten times. Then airway hyperresponsiveness, inflammatory cells and eicosanoids in the bronchoalveolar lavage fluid (BALF) and peripheral blood were examined. Airway responsiveness was acquired after inhalation of the antigen. Two types of airway responsiveness were observed ; accerelating-group and recovering-group. Total cell count and numbers of eosinophils and neutrophils in the BALF were significantly increased in the accerelating-group compared with the recovering-group and the control-group. The TXB2 level in the BALF was increased in the accerelating-group, and the LTB4 level in the BALF and peripheral blood were also increased in the accerelating-group. These findings suggest that the airway hyperresponsiveness is caused by eicosanoids, such as thromboxanes and leukotrienes produced by some inflammatory cells which are organized by activated lymphocytes as well as mast cell-basophils.

呼吸器疾患におけるステロイドの効果の指標として家兎気管支肺胞洗浄細胞(以下BAL-cell)のグルココルチコイドレセプター(以下GR)をとりあげ,宿主因子(加齢,活性化,絶食)およびステロイド投与が及ぼす変化を観察した.その結果,加齢では若年群ほど高値を示した. 絶食では一過性に増加した後,減少傾向を示し,さらにheat-killed BCGによって活性化されたBAL-cellのGRの上昇を抑制した.一方,プレドニゾロン1回投与によるGRの推移をheat-killed BCG感作群と非感作群において観察したところ,両群とも3時間後に低下し, 2日から4日後に一過性に上昇を示したが7日後に前値に復した. 以上の結果より,ステロイド治療を施行する際は,これらの宿主因子およびステロイド投与がBAL-cellのGRを変化させることを十分考慮に入れる必要があると考えられた

The bronchoalveolar lavage (BAL) fluid of patients with PIE syndrome characterized by pulmonary eosinophilia was examined to determined the cellular response in the lungs. Evaluation of cellular components in the BAL fluid revealed an increased proportion not only of eosinophils but also of lymphocytes and neutrophils. In about half of the patients with PIE syndrome the level of eosinophilia was higher in the peripheral blood than in the BAL fluid. Patients with PIE syndrome were classified into prolonged PIE and PIE with asthma based on Crofton's classification. The percentage of neutorphils and eosinophils in BAL fluid were higher in patients with prolonged PIE than in PIE with asthma while the percentage of lymphocytes was higher in a group of PIE with asthma. On the other hand, the lymphocyte precentage in BAL fluid was higher in patients with PIE syndrome due to fungus antigen than in those with PIE syndrome due to drug allergy. These findings suggest that various cellular components play important roles in the pathogenesis of PIE syndrome and that the accumulation of the effector cells in the lungs is regulated by an allergic mechamism.

To clarify the mechanism of late asthmatic response (LAR) which is related to intractable asthmatic attacks, cellular and humoral components at the sites of LAR were examined by bronchoalveolar lavage fluid (BALF) obtained after bronchial inhalation of Candida allergen. BAL examination was performed 2 hours after the LAR attack. The differential cell count was obtained and the level of leukotrienes (LTs) in the BALF and peripheral blood determined by high performance liquid chromatography. The percentages of eosinophils and basophil・mast cells and neutrophils were statistically high in the BALF after LAR compared with the level in the non-attack state (p

To investigate the mechanism of the late asthmatic response (LAR) which participates in intractable asthmatic attacks, humoral and cellular components at the sites of LAR were examined by bronchoalveolar lavage fluid (BALF) obtained after bronchial inhalation of house dust allergen. BAL examination was performed 2 hours after improvement of LAR, and leukotrienes (LTs) measured by high performance liquid chromatography. The level of LTC4 was significantly higher in BALF after LAR(p

Various allergic and immunologic mechanisms have been postulated in the pathogenesis of PIE syndrome. In this study, the immune response to Aspergillus antigen was examined in patients with the PIE syndrome of allergic bronchopulmonary aspergillosis (ABPA) and those with aspergilloma to elucidate the immune mechanisms wihch regulate the pulmonary changes in PIE syndrome. The same Aspergillus species causes different clinical disease entities such as Aspergillus pneumonia, aspergilloma, ABPA and even hypersensitivity pneumonitis. The specific IgE against the Aspergillus antigen for the type Ⅰ allergy existed in patients with ABPA but not in those with Aspergilloma. However, the prepicitating antibody for the type Ⅲ allergy and enhanced lymphocyte blastogenesis against Aspergillus antigen for the type Ⅳ allergy were found in both the patients with ABPA and those with aspergilloma while pulmonary infiltration, dyspnea and the grade of peripheral eosinophilia correlated with the level of serum IgE and responsiveness of lymphocyte against Aspergillus antigen. These findings indicate that the characteristics of the immune responses of the host play a crucial role in the pathogenesis of PIE syndrome.

Abnormality of cellular immunity has been reported in the pathogenesis of collagen vascular diseases (CVD). IL-2 and IL-2 receptors play important roles as immunological regulation in CVD. In this study, the soluble forms of IL-2 receptor (sIL-2R) and IL-2 receptor positive lymphocytes (CD25) were analyzed to evaluate the role of IL-2R in the pathogenesis of interstitial pneumonia (IP) of CVD such as RA, SLE, polymyositis/dermatomyositis (PM/DM), and PSS and idiopathic interstitial pneumonia (IIP) as a disease control. High levels of sIL-2R were found in sera of patients with IP of CVD. The level of soluble IL-2R in sera of RA without IP correlated with the disease activity of the joints, while the level of sIL-2R in sera of RA with IP was higher than in that of RA without IP. Soluble IL-2R in bronchoalveolar lavage fluind (BALF) was also measured and the level was higher in IP with CVD. There were fewer IL-2R positive lymphocytes in the peripheral blood of CVD with IP than in the normal controls or IIP cases, although the serum level of sIL-2R was higher in CVD with IP and IIP than in the normal controls. These findings indicate that actibve immunological activities involving IL-2/IL-2R/sIL-2R systems play an important role in the pathogenesis of IP of CVD.

The cellular components in lungs of idiopathic interstitial pneumonia (IIP) and interstitial pneumonia with collagen disease (IP with CD) were examined by bronchoalveolar lavage (BAL) to analyze the pathogenesis of IIP. The findings of BAL fluid of IIP and IP with CD were increase of total cell count and increased percent of lymphocytes, neutrophils, eosinophils and basophil/mast cells in comparison with the normal control. The BAL fluid of the patients with IIP and IP with CD in active stage contained a higher proportion of lymphocyte, neutrophil, eosinophil and basophil/mast cell than those in a nonactive stage. Furthermore, an increase of T-cells and Ia positive T-cells was detected in the examination of lymphocyte subpopulation in BAL fluid of patients with IIP, while the lymphocyte subpopulation of peripheral blood showed no significant difference. Cellular components in BAL fluid of IIP patients treated with or without steroids were compared. Under steroid therapy a decrease of total cell count and eosinophil was shown in BAL fluids of patients with IIP. These findings indicate that various cells such as macrophage, lymphocyte, neutrophil eosinophil and basophil/mast cell in lungs play important roles in the pathogenesis of IIP and IP with CD.

Various pulmonary involvements have been reported in collagen vascular diseases (CVD). Interstitial pneumonia (IP) is one of the common complications. The pathogenesis of IP is still obscure. In this study bronchoalveolar lavage (BAL) was applied to analyze the cellular response in the pathological changes in the bronchioloalveolar regions in IP of CVD such as RA, SLE, polymyositis/dermatomyositis and PSS. Increased total cell counts and cellular density in the BAL fluid were shown in most of the CVD with IP. Furthermore, a relative decrease in the percetn of macrophages and increase in the percent of lymphocytes, neutrophils, eosinophils and basophil/mast cells in BALL fluid were revealed, the increase being more marked in active IP than in non-active IP. T-cell subsetes of lymphocytes in the BALL fluid were analyzed with flow cytometry. A decrease in the CD4/CD8 ratio was shown in most of the IP with CVD except for RA in which the CD4/CD8 ratio was increased. Total cell count and percentage of macrophage was higher in the smoking group than in the non-smoking group of IP with RA. These findings indicate that cellular responses obtained by BAL could be useful not only for analyzing the pathogenesis of IP of CVD but also for evaluating the disease activity of IP.

Radiation pneumonitis （RP） is a serious disorder caused by radiation therapy, for the detection of which there exists no simple and sensitive clinical examination. In this study, we examined whether the measurement of serum pulmonary surfactant protein A （SP-A） levels could be effective for detecting the onset of RP. Of the 18 patients included in the study, 9 suffered RP complication after radiation therapy. Serum SP-A levels in patients with RP （64.1±6.5 ng/ml） were 1.82 times as high as those of pre-radiation （36.2±4.0 ng/ml）. The difference between them was significant （p=0.0003）, while the SP-A value in patients without RP after radiation therapy was almost the same as the pre radiation level. To study the mechanisms of these elevations, we developed a model of whole lung irradiation （20Gy） in adult rats. Prior to alveolitis, which was a histological change seen at day 28, levels of SP-A in lung tissue homogenates and bronchoalveolar lavage fluid had become elevated by day 21. Serum SP-A levels in the irradiated rats were significantly elevated by day 21, peaking at day 35, and then declined. Hydroxyproline contents in lung tissues were elevated by day 42 and remained so through day 56. Therefore, the increase in serum SP-A level may be reflected by alveolitis but not fibrosis. In conclusion, determination of serum SP-A could be helpful in detecting the onset of RP.

Fibronectin (FN) is known to promote fibroblast proliferation and attachment of collagen fibers at the site of tissue repair or fibrosis. Although increaed levels of FN were shown in bronchoalveolar lavage (BAL) fluid of patients with interstitial pneumonia, the source of FN and the role of FN in the pathogenesis of interstitial pneumonia are still obscure. BAL fluid of patients with various types of interstitial pneumonia were obtained to measure FN and albumin (Alb) levels and to examine the cellular components. Because of the different recovery rate of BAL fluid, the FN/Alb ratio of the BAL fluid was evaluated as the real FN level in the lungs. A high FN/Alb ratio was shown in patients with idiopathic interstitial pneumonia, interstitial pneumonia with rheumatoid arthritis, hypersensitivity pneumonitis, and sarcoidosis in comparison with normal volunteers. Patients with impaired pulmonary function or acute and progressive interstitial pneumonia were shown to have a higher FN/Alb ratio than patients with normal pulmonary function or inactive interstitial penumonia. The levels of FN in BAL fuid showed a statistically significant correlation with total cell, alveolar macrophage and neutrophil concentrations of BAL fluid, but not with the lymphocyte concentration. Alveolar macrophages, isolated as adhesive cells by a brief incubation in plastic culture plates, were cultured for 48 hours, and the level of FN in the culture medium was measured. FN production by alveolar macrophages was greater in patients with interstitial pneumonia than in norman volunteers. These data suggest that the local increased production of FN might accelerate the fibrotic process in the alveolar wall, following the accumulation of alveolar macrophages and other cellular components.

Gallium-67 citrate ((67)Ga) is known to accumulate in inflammatory areas. Though the exact mechanism of the transportation and localization of (67)Ga citrate in foci of inflammaion is still obscure, (67)Ga scintgraphy has been used clinically to evaluate patients with interstitial pneumonia with regard to disease activity and progression. Quantitative analysis using (67)Ga scintigraphy has been attempted several times, but the results have been inadequate, even with computer analysis. In this study, the combination of (67)Ga scintigraphy and bronchoalveolar lavage (BAL) was applied to patients with various types of interstitial pneumonia to estimate quantatively (67)Ga accumlation in the lungs. The radio activity of BAL fluid and peripheral blood were measured directly, and the ratio of radioactivity of BAL fluid/peripheral blood was calculated for each of the patients. Radioactivity in the peripheral blood was contained mainly in the plasma. On the other hand, most radioactivity in BAL fluid was shown to be located in the cellular component. The cellular component of BAL fluid was separated by culture in plastic culture plates into adherent cells (macrophage-enriched fraction) and non-adherent cells (lymphocyte-enriched fraction). Comparison of radioactivity of each cell fraction proved that alveolar macrophages contained most of the (67)Ga citrate in BAL fluid. Therefore, the high (67)Ga uptake in scintigrams of patients with interstitial pneumonia represents the increased number or activated state of alveolar macrophages in the lungs. The direct measurement of (67)Ga radioactivity in BAL fluid could be a useful method to analyze quantatively (67)Ga scintigrams of interstitial pneumonia patients.

Cytological studies were performed on bronchoalveolar lavage(BAL) fluid in 7 cases with hypersensitivity pneumonitis, 6 with sarcoidosis, 6 with chronic bronchitis and 5 with bronchiolitis. In the cases with hypersensitivity pneumonitis and bronchiolitis, cytological studies of BAL fluid were compared with the histological findings of lungs. Furthermore, animal models of hypersensitivity pneumonitis were made and examined likewise. The results were as follows. 1) In the cases with hypersensitivity pneumonitis, a marked increase in the lymphocyte count in BAL fluid was characteristic. These cases showed an increase in the percentage of T lymphocytes in BAL fluid as compared with healthy subjects. In the cases with sarcoidosis, an increase in the number of lymphocytes was also found and especially the ratio of the percentage of T lymphocytes to that of B lymphocytes was much higher than those of the other diseases and healthy subjects. 2) An increased number of neutrophils in BAL fluid was observed in the cases with chronic bronchitis. In the cases with bronchiolitis, a marked increase in the neutrophil count was characteristic. 3) A common histological finding in hypersensitivity pneumonitis was mononuclear cell infiltration into alveolar septa and most of the cells in BAL fluid were lymphocytes. In bronchiolitis, mononuclear cell infiltration into and around the bronchiolar walls was found commonly and neutrophils were found increased in BAL fluid. 4) In experimental hypersensitivity pneumonitis, mononuclear cell infiltration into septa closely paralleled the lymphocyte appearance in BAL fluid.

Sarcoidosis is a systemic disorder characterized by the accumulation of a large number of T-lymphocytes in pulmonary sites. Considerable evidence has been presented that alveolar lymphocytes are activated by some agent in pulmonary sarcoidosis, although the pathogenesis of this disease remains unknown.The proliferation of lymphocytes, obtained by bronchoalveolar lavage, was determined by in vitro incorporation of 3H-thymidine induced by Propionibacterium acnes (P.acnes).The mean response rate of alveolar lymphocytes was 1.91±0.80 in 22 untreated sarcoidosis patients, 1.22±0.46 in nine sarcoidosis patients given prednisolone, and 0.90±0.39 in 15 controls. The response rate was significantly enhanced in untreated patients compared to both treated patients (P

T-lymphocyte subsets in bronchoalveolar lavage fluid (BALF) were examined to study immunological abnormalities in the lung of bronchial asthma patients. T-cell subsets were determined by immunofluorescent microscopy using OKT monoclonal antibodies: T3 for pan T-cells, T4 for helper/inducer cells, and T8 for suppressor/cytotoxic cells. In these analyses, patients with bronchial asthma were classified according to the background of atopy, onset age, and duration of asthmatic symptoms. Decreased T4 percent and T4/T8 ratio were found in patients with non-atopic asthma, patients with onset later than 40 years of age, and patients with a duration of less than 5 years. Although corticosteroids are known to decrease the T4 percent and T4/T8 ratio in peripheral blood lymphocytes, their effect on T-cell subsets in BALF is not well known. In this study, the T4 percent and T4/T8 ratio in BALF were found to be lower in patients with corticosteroids than in patients without corticosteroids and in healthy subjects. These results indicate that an immunological abnormality which might be related to the pathogenesis of bronchial asthma exists in particular groups of asthmatic patients.

Bronchoalveolar lavage and subsequent bronchial brushing techniques were examined in 60 consecutive cases of endoscopically non-visible peripheral lung cancer in an attempt to improve the diagnostic rate. Lavaged fluid was processed routinely for cytological examinations, and carcinoembryonic antigen (CEA) concentrations were measured concomitantly. Lavaged fluid gave a cytological diagnosis in 63.3% of the patients. The positive cytology rate by brushing was 76.7% . Of 14 patients with negative cytology on brushing, 6 were positive in lavaged fluid. The combination of bronchoalveolar lavage and subsequent bronchial brushing gave a cytological diagnosis in 86.7% of the patients. CEA concentrations of lavaged fluid of patients with lung cancer were higher than those of patients with non-malignant pulmonary diseases. It may be said that increased CEA concentrations in lavaged fluid suggest lung cancer. The relatively low positive rate (76.7%) by brushing cytology appeared due to a technical problem that the preceding bronchoalveolar lavage made it difficult to accomplish a precise brushing under fluoroscopy. Accordingly, the sequence of the two techniques was reversed thereafter. Of 40 consecutive cases with endoscopically non-visible peripheral lung cancer, brushing cytology gave a diagnosis in 87.5% , and bronchoalveolar lavage did so in 65.0% . The combination of bronchial brushing and subsequent bronchoalveolar lavage gave a cytological diagnosis in 92.5% of the patients. In conclusion, bronchoalveolar lavage was useful in the diagnosis of patients with peripheral lung cancer.

Cytological studies were performed on bronchoalvelar lavage(BAL) fluid in 40 cases with bronchial asthma. The results were as follows. 1) In 13 healthy subjects the cells in BAL fluid comprised 89.6% macrophages, 9.7% lymphocytes, 0.7% neutrophils, 0.4% eosinophils and 0% basophils. 2) In asthmatic cases the cells in BAL fluid comprised 54.4% macrophages, 12.6% lymphocytes, 11.8% neutrophils, 21.2% eosinophils and 0.1% basophils. These cases showed a marked increase in the number of eosinophils and a moderate increase in the neutrophil count as compared with healthy subjects. The increased number of eosinophils in lavage fluid was relatively closely related to positive immediate skin tests, high serum IgE levels and peripheral eosinophilia. As for asthmatic types, this was marked in the atopic type. By contrast, an increased number of neutrophils in the lavage fluid was frequently found in non-atopic or intractable asthmatics showing the frequent complication of peripheral airway disorder in pulmonary function tests. Therefore, it seems that the present study gave much information on the characteristics and the etiology of the intractable asthmatics. 3) Eosinophils in BAL fluid reflected the appearance of eosinophils in the alveolar spaces and eosinophil infiltration into the septa in the histological findings. On the other hand, neutrophils could be observed clearly by the BAL method even when they could not be observed clearly by histological studies. In conclusion, the cytological studies performed on BAL fluid are considered useful in the diagnosis of bronchial asthma, especially for the classification into subtypes of asthma.

The present study was designed to investigate the local immunity and pathological state of sarcoidosis. Cell populations and angiotensin converting enzyme (ACE) activity were examined in the fluid obtained by bronchoalveolar lavage (BAL) of 28 patients with sarcoidosis and 31 healthy volunteers. The proportion and number of lymphocytes in BAL fluid were significantly higher in patients with sarcoidosis than in healthy volunteers. Furthermore, the number of T lymphocytes and B lymphocytes were increased. The cell number in BAL fluid was also higher in non-smokers with sarcoidosis than in healthy non-smokers. The ACE activity in bronchoalveolar lavage fluid (BALF-ACE activity) was highly elevated in patients with sarcoidosis as compared with healthy volunteers and correlated with the number of macrophages in BAL fluid of non-smokers with sarcoidosis (r=0.474, p

肺胞マクロファージは,常に外気と接触し刺激を受けている細胞であり肺胞内細胞の90%以上を占め,生体防御機構において重要な働きをしているものと考えられている。また肺内における環境の変化に対応して腫揚免疫においても重要なエフェクター細胞と考えられる。しかし,肺癌の発癌過程における肺胞マクロファージの免疫学的意義は,十分に明らかではない。そこで著者は家兎を用い,肺胞マクロファージの機能的変化を,抗腫瘍活性を中心に検討した。 まず,家兎の肺胞マクロファージは,気管支ファイバースコープにてBALを行う事により採取したが,はじめにその方法,間隔について検討した。次に家兎の肺に化学発癌を惹起させ,発癌過程における肺胞マクロファージの機能的変化を観察した。さらにBRMを用いその抗腫瘍活性の増強方法について検討し,以下の結論を得た。 1) 家兎における,気管支ファイバースコープを用いたBALは,注入量20ml,洗浄回数2回で2～3×10個の肺胞マクロファージを回収できた。 2) BALの施行間隔を1週間以下とした場合,肺胞内細胞成分または肺胞マクロファージの機能に変化が認められた。2週間では変化を認めなかった。 3) 家兎肺胞マクロファージは,6種のヒト培養腫瘍細胞株に対し,増殖抑制能を示したが,正常細胞であるヒト線維芽細胞に対しては,増殖抑制能を示さなかった。 4) MNUを左主気管支に注入し, 12週で1羽/3羽, 24週で3羽/3羽の中枢型肺癌の発生を認めた。 5) その発癌過程における肺胞マクロファージの腫瘍細胞増殖抑制能を観察し,経時的な上昇を認めた。 6) 実験12週で屠殺した2羽の家兎において,発癌家兎と非発癌家兎の間で腫瘍細胞増殖抑制能の上昇の程度には差を認めなかった。実験24週で屠殺した3羽の家兎において癌腫の浸潤の程度と腫瘍細胞増殖抑制能の上昇に関連は認められなかった。 7) 発癌過程における肺胞マクロファージのNBT還元能は有意な変化を示さず,貪食能は経時的な低下,免疫抑制活性は経時的な上昇を認めた。免疫抑制活性は個体差を認めた。 8) 発癌過程における肺胞マクロファージの腫癌細胞増殖抑制能は,免疫抑制活性との間に正の相関を認め,貪食能との問には負の相関を認めた。 9) LPS, N-CWS, IFNγは,腫瘍細胞に対し,直接の増殖抑制作用は示さず, TNFは腫蕩細胞に対し,腫瘍細胞の種頬によっては直接,増殖抑制作用を示した。 10) LPS, N-CWS, IFNγは,それぞれ肺胞マクロファージの腫瘍細胞増殖抑制能を増強させた。 11) その場合IFNγに対する反応性は,個体差が認められ反応性を示さないものも認められた。 12) 反応性を示さない家兎においてもIFNγ添加後にLPSを添加した場合は, IFNyに容量依存性作用の発現を認めた。 13) 併用の場合他に, N-CWSとIFNγ, N-CWSとTNFおよびIFNyとTNFの組み合わせにおいて肺胞マクロファージの腫瘍細胞増殖抑制能増強作用に関して相乗効果が認められた。TNFを含む組み合わせでは,特にTNF resistant cell lineに対して強い相乗効果を認めた。 以上より肺胞マクロファージは発癌の初期段階において抗腫瘍性を上昇させており,それはBRMの併用により効果的に上昇させ得ることが試験管内において確認できた。但し腫瘍細胞増殖抑制能と免疫抑制活性の間には相関が認められ肺胞マクロファージの抗腫癌性の検討の際に注意が必要であると思われた。, To examine the effects of carcinogenesis on the antitumor activity of alveolar macrophages, MNU was instilled into the rabbit bronchus to produce cancer, and changes in rabbits were observed for the next 24 weeks. Alveolar samples were collected by bronchofiberscopic BAL (Bronchoalveolar lavage), which does not require animal sacrifice. Although BAL itself causes changes in the lung, it was confirmed an interval of two weeks allows BAL to be performed serially. Significant changes with time in the total cell count and differential cell count in BALF (Bronchoalveolar lavage fluid) were not seen. Functions of PAM were as follows: cytostatic activity increased progressively, phagocytosis gradually decreased, and both showed significant differences from pre-values. No significant change with time was noted m the NBT reduction. Suppressor activity increased with time in some rabbits, and showed a significant correlation with cytostatic activity. The in vitro effect of BRMs on the cytostatic activity of alveolar macrophage was also studied. LPS potenbated the cytostatic activity against each of 3 kinds of target cells, while N-CWS, IFN and TNF each potentiated activity partially. When these were used in combinations, synergistic effects were observed between LPS and IFN, between N-CWS and IFN and between TNF and N-CWS., 広島大学医学雑誌, 37(3), 347-389, 平1-6月 (1989)

肺胞マクロファージは,常に外気と接触し刺激を受けている細胞であり肺胞内細胞の90%以上を占め,生体防御機構において重要な働きをしているものと考えられている。また肺内における環境の変化に対応して腫揚免疫においても重要なエフェクター細胞と考えられる。しかし,肺癌の発癌過程における肺胞マクロファージの免疫学的意義は,十分に明らかではない。そこで著者は家兎を用い,肺胞マクロファージの機能的変化を,抗腫瘍活性を中心に検討した。 まず,家兎の肺胞マクロファージは,気管支ファイバースコープにてBALを行う事により採取したが,はじめにその方法,間隔について検討した。次に家兎の肺に化学発癌を惹起させ,発癌過程における肺胞マクロファージの機能的変化を観察した。さらにBRMを用いその抗腫瘍活性の増強方法について検討し,以下の結論を得た。 1) 家兎における,気管支ファイバースコープを用いたBALは,注入量20ml,洗浄回数2回で2～3×10個の肺胞マクロファージを回収できた。 2) BALの施行間隔を1週間以下とした場合,肺胞内細胞成分または肺胞マクロファージの機能に変化が認められた。2週間では変化を認めなかった。 3) 家兎肺胞マクロファージは,6種のヒト培養腫瘍細胞株に対し,増殖抑制能を示したが,正常細胞であるヒト線維芽細胞に対しては,増殖抑制能を示さなかった。 4) MNUを左主気管支に注入し, 12週で1羽/3羽, 24週で3羽/3羽の中枢型肺癌の発生を認めた。 5) その発癌過程における肺胞マクロファージの腫瘍細胞増殖抑制能を観察し,経時的な上昇を認めた。 6) 実験12週で屠殺した2羽の家兎において,発癌家兎と非発癌家兎の間で腫瘍細胞増殖抑制能の上昇の程度には差を認めなかった。実験24週で屠殺した3羽の家兎において癌腫の浸潤の程度と腫瘍細胞増殖抑制能の上昇に関連は認められなかった。 7) 発癌過程における肺胞マクロファージのNBT還元能は有意な変化を示さず,貪食能は経時的な低下,免疫抑制活性は経時的な上昇を認めた。免疫抑制活性は個体差を認めた。 8) 発癌過程における肺胞マクロファージの腫癌細胞増殖抑制能は,免疫抑制活性との間に正の相関を認め,貪食能との問には負の相関を認めた。 9) LPS, N-CWS, IFNγは,腫瘍細胞に対し,直接の増殖抑制作用は示さず, TNFは腫蕩細胞に対し,腫瘍細胞の種頬によっては直接,増殖抑制作用を示した。 10) LPS, N-CWS, IFNγは,それぞれ肺胞マクロファージの腫瘍細胞増殖抑制能を増強させた。 11) その場合IFNγに対する反応性は,個体差が認められ反応性を示さないものも認められた。 12) 反応性を示さない家兎においてもIFNγ添加後にLPSを添加した場合は, IFNyに容量依存性作用の発現を認めた。 13) 併用の場合他に, N-CWSとIFNγ, N-CWSとTNFおよびIFNyとTNFの組み合わせにおいて肺胞マクロファージの腫瘍細胞増殖抑制能増強作用に関して相乗効果が認められた。TNFを含む組み合わせでは,特にTNF resistant cell lineに対して強い相乗効果を認めた。 以上より肺胞マクロファージは発癌の初期段階において抗腫瘍性を上昇させており,それはBRMの併用により効果的に上昇させ得ることが試験管内において確認できた。但し腫瘍細胞増殖抑制能と免疫抑制活性の間には相関が認められ肺胞マクロファージの抗腫癌性の検討の際に注意が必要であると思われた。, To examine the effects of carcinogenesis on the antitumor activity of alveolar macrophages, MNU was instilled into the rabbit bronchus to produce cancer, and changes in rabbits were observed for the next 24 weeks. Alveolar samples were collected by bronchofiberscopic BAL (Bronchoalveolar lavage), which does not require animal sacrifice. Although BAL itself causes changes in the lung, it was confirmed an interval of two weeks allows BAL to be performed serially. Significant changes with time in the total cell count and differential cell count in BALF (Bronchoalveolar lavage fluid) were not seen. Functions of PAM were as follows: cytostatic activity increased progressively, phagocytosis gradually decreased, and both showed significant differences from pre-values. No significant change with time was noted m the NBT reduction. Suppressor activity increased with time in some rabbits, and showed a significant correlation with cytostatic activity. The in vitro effect of BRMs on the cytostatic activity of alveolar macrophage was also studied. LPS potenbated the cytostatic activity against each of 3 kinds of target cells, while N-CWS, IFN and TNF each potentiated activity partially. When these were used in combinations, synergistic effects were observed between LPS and IFN, between N-CWS and IFN and between TNF and N-CWS., 広島大学医学雑誌, 37(3), 347-389, 平1-6月 (1989)