结节病患者肺功能与支气管肺泡灌洗液细胞学关系的研究

目的 探讨结节病患者不同影像分期之间肺功能指标、支气管肺泡灌洗液细胞学的改变以及两者的相关性.方法 回顾性调查71例结节病患者肺功能以及支气管肺泡灌洗液检查的资料.结果 在结节病患者不同影像分期之间肺功能指标用力肺活量(FVC)占预计值%、第1秒用力呼气容积(FEV1)占预计值%、肺总量(TLC)占预计值%以及肺一氧化...     

Uniformity of bronchoalveolar lavage in patients with pulmonary sarcoidosis

Sarcoidosis is a granulomatous disease of unknown cause characterized by a lymphocytic alveolitis. Previous studies have shown that the inflammatory cell population of the distal lung units of patients with this disorder can be accurately assessed using bronchoalveolar lavage (BAL). The present study evaluated the uniformity of BAL between different sites of the lung in patients with sarcoidosis. In general, there was a good correlation between sites for percentages of lymphocytes (LYM) (r = 0.750, p less than 0.0001), LYM number (r = 0.356, p = 0.0007), percentages of neutrophils (NEUT) (r = 0.917, p less than 0.0001), NEUT number (r = 0.999, p less than 0.0001), and macrophage (MAC) number (r = 0.858, p less than 0.001). Despite the good overall correlation, we found that 43% of the patients with high percent LYM (greater than 30%) had this finding on one side only. These patients did not differ from the group as a whole based on radiographic stage of their disease but did differ in the number of radiographs demonstrating focal infiltrates (2 of 28 patients with both sides less than 30% LYM, 2 of 14 with both sides greater than 30% LYM, and 4 of 9 with only one side greater than 30% LYM p less than 0.05 by chi-square); and in each situation the highest percent LYM was seen on the side with focal changes on the chest radiograph.(ABSTRACT TRUNCATED AT 250 WORDS)     

High concentrations of soluble Fas ligand in bronchoalveolar lavage fluid of patients with pulmonary sarcoidosis

BACKGROUND AND OBJECTIVES: Sarcoidosis is a systematic granulomatous disorder of unknown origin characterized by accumulation of T lymphocytes and macrophages in multiple organs. We postulated that apoptosis through the Fas/Fas ligand (L) system may be associated with regulation of immune reactions characterized by the formation of noncaseous necrotizing granulomas. Soluble (s) FasL is not equivalent to membrane-associated FasL since conversion of membrane-bound FasL to the soluble form is associated with downregulation of cytotoxicity. To examine the involvement of sFasL in lung inflammation, we compared the levels of sFasL in bronchoalveolar lavage (BAL) fluid and serum of patients with pulmonary sarcoidosis to those of healthy subjects. METHODS: sFasL was measured in BAL fluid and in serum of 15 patients with active pulmonary sarcoidosis by sandwich ELISA. RESULTS: High concentrations of sFasL were detected in BAL fluid and serum of patients with sarcoidosis but not in normal subjects. There was a significant correlation between the percentage of lymphocytes and sFasL concentrations in BAL fluid (r = 0.585, p < 0.05). CONCLUSIONS: Our results suggest that sFasL may be upregulated locally in the lung during the inflammatory process of active pulmonary sarcoidosis.     

KL-6, surfactant protein A and D in bronchoalveolar lavage fluid from patients with pulmonary sarcoidosis.

BACKGROUND: KL-6, and surfactant protein A (SP-A) and surfactant protein D (SP-D) derived from alveolar type II cells and/or bronchiolar epithelial cells have been reported to be useful markers for interstitial lung diseases. OBJECTIVE: The aim of this study was to measure the levels of these molecules in bronchoalveolar lavage fluid (BALF) from patients with pulmonary sarcoidosis to investigate their relationship with other markers of inflammatory activity. METHODS: We measured KL-6, SP-A and SP-D levels in BALF from patients with pulmonary sarcoidosis using an ELISA. RESULTS: KL-6 and SP-D, but not SP-A levels were significantly increased in pulmonary sarcoidosis compared with controls. KL-6, SP-A and SP-D levels were significantly correlated with each other. KL-6 and SP-D levels were relatively and significantly correlated with the percentage of lymphocytes in BALF. KL-6, SP-D, but not SP-A levels were significantly correlated with the concentration of albumin in BALF. There was no significant correlation between KL-6, SP-A, or SP-D levels and chest X-ray findings, angiotensin-converting enzyme levels, or CD4/CD8 ratio in BALF. CONCLUSIONS: We conclude that KL-6 and SP-D levels in BALF were increased in pulmonary sarcoidosis. Since these markers are specifically derived from epithelial cells, it is considered that KL-6 and SP-D levels are reflecting damage or release of these markers from epithelial cells due to the inflammatory response.     

Phagocyte enzymes in bronchoalveolar lavage from patients with pulmonary sarcoidosis and collagen vascular disorders

The balance between proteases and antiproteases in the lower respiratory tract is believed to play a role in the outcome of interstitial lung diseases. In this cross-sectional study, we measure several phagocyte derived enzymes, namely plasminogen activator, neutrophil elastase and an ill-defined protease active on the trialanine chromophore substrate succinyl-alanine3-nitroanilide (SLAPN) in bronchoalveolar lavage (BAL) fluid from 42 patients with pulmonary sarcoidosis and from 43 patients with collagen vascular disease (CVD), 22 without lung disease (group I) and 21 associated with parenchymal lung disease (group II). The results show: a) that sarcoidosis is associated with increased plasminogen activator activity and with the presence of enzymatic activity against SLAPN corresponding at least in part to a metalloprotease; b) that CVD in the absence of radiographic lung disease is associated with an increase of plasminogen activator activity and increased levels of alpha 1-antiprotease-neutrophil elastase complexes; c) that the majority of untreated CVD (group II) patients have detectable levels of neutrophil elastase activity. These data show that patients with pulmonary sarcoidosis and CVD have different enzymatic profiles in their lower respiratory tract as assessed by BAL. Thus, sarcoidosis (mostly lymphocytic) is associated with enhanced macrophage-derived proteolytic activity in BAL, while CVD patients both with and without lung disease have increased neutrophil counts and neutrophil elastase complexed to alpha 1-protease inhibitor and presumably inactive in BAL. Finally, only BAL from untreated CVD patients with interstitial lung disease contain neutrophil elastase activity. This latter activity could contribute to the lung lesions frequently observed in these disorders.     

Antibacterial components in bronchoalveolar lavage fluid from healthy individuals and sarcoidosis patients.

Antibacterial peptides and proteins are an integral part of the epithelial defense barrier that provides immediate protection against bacterial invasion. In humans, alpha-defensins are mainly bactericidal effectors in circulating granulocytes, beta-defensin-1 is synthesized in epithelial cells, and LL-37 is produced in granulocytes but is also induced in skin epithelia during inflammation. To investigate the importance of these defense effectors in disease, we analyzed bronchoalveolar lavage fluid (BALF) for bactericidal activity. Antibacterial activity was found in BALF material from healthy individuals and sarcoidosis patients, with enhanced activity in BALF from the patients. The activity was present as several antibacterial components, of which we have so far characterized LL-37, lysozyme, alpha-defensins, and antileukoprotease. In addition, the antibacterial peptide LL-37 was located in alveolar macrophages, bronchial epithelial cells, and bronchial glands, suggesting that it has a defensive role in airway mucosa. In conclusion, the airway epithelium is protected by a complex antibacterial defense system. This is activated in sarcoidosis, and may explain why these patients seldom develop severe respiratory tract infections.     

Platelet-activating factor in bronchoalveolar lavage from patients with sarcoidosis.

Platelet-activating factor (PAF), a lipid mediator of inflammation and anaphylaxis, may play a role in several physiopathologic alterations of the lung. A lipid compound with physicochemical and biologic characteristics similar to synthetic PAF was extracted and purified from bronchoalveolar lavage (BAL) fluid of 15 of 34 patients with sarcoidosis. PAF was quantitated by a bioassay on washed rabbit platelets. The specificity of platelet aggregation was assessed by using two different PAF receptor antagonists. The incidence of detectable amounts of PAF in BAL fluid of sarcoid patients was statistically significant (chi 2 = 4.064, p = 0.044) when compared with the 14 normal control subjects. The results demonstrated an increased production of PAF in the lower respiratory tract of patients with sarcoidosis. The presence of PAF in BAL fluid, however, did not correlate with radiologic stage, intensity of alveolitis, gallium scanning positivity, angiotensin-converting enzyme serum level, or lung function tests. Therefore, a direct relationship between presence of PAF in BAL fluid and activity of lung disease in patients with sarcoidosis was not directly established.     

Studies of bronchoalveolar lavage cells and fluids in pulmonary sarcoidosis. I. Enhanced capacity of bronchoalveolar lavage cells from patients with pulmonary sarcoidosis to induce angiogenesis in vivo

Both allogeneic immunocompetent CD4+ lymphocytes and activated macrophages of mice can induce neovascularization when inoculated intradermally into host animals. Because sarcoidosis is associated with an increase in both activated macrophages and CD4+ effector lymphocytes in the lung, we carried out experiments in which cells obtained by bronchoalveolar lavage (BAL) of patients with pulmonary sarcoidosis were tested in a murine intradermal angiogenesis assay. BAL cells from patients with pulmonary sarcoidosis induced a significantly greater degree of angiogenesis than those from normal volunteers or from patients with other lung diseases. Moreover, the degree of angiogenesis induced by BAL cells from patients with sarcoidosis correlated positively with the severity of the disease. When BAL cells were separated into macrophage and lymphocyte subpopulations by flow cytometric techniques, the observed angiogenic activity was restricted primarily or exclusively to macrophages; lymphocytes were unable to induce angiogenesis in this xenogeneic assay system. These experiments suggest that pulmonary macrophages may play a role in the pathogenesis of sarcoidosis by inducing changes in the pulmonary microvasculature. Moreover, we hypothesize that these vascular changes may be induced not only in the lung but also in other organ systems such as skin, muscle, and eye in which microangiopathies are associated with sarcoid disease.     

Adenosine deaminase activity in bronchoalveolar lavage fluid of sarcoidosis patients]

We studied adenosine deaminase (ADA) activity in bronchoalveolar lavage fluid (BALF) specimens from 24 patients with sarcoidosis. Mean BALF-ADA activity was significantly (p < 0.0001) elevated in patients with sarcoidosis (1.02 +/- 1.01 IU/L, mean +/- SD) compared to the subjects in a healthy control group (0.08 +/- 0.29 IU/L). In the sarcoidosis patients with high BALF-ADA activity (> or = 1.0 IU/L), AaDO2 was significantly (p < 0.0001) elevated compared to its level in patients with normal BALF-ADA activity (< 1.0 IU/L). BALF-ADA activity was significantly (p < 0.01) higher in patients who exhibited lung-field accumulations on 67Ga scintigrams compared to those with no accumulations, and significantly (p < 0.001) higher in patients undergoing corticosteroid treatment compared to in those patients who did not receive such treatment. These findings were similar to the results of studies using data on BALF-ADA/albumin ratios. Furthermore, they suggest that the localized production of ADA may increase in sarcoidosis patients displaying 67Ga scintigram lung-field accumulations with increased AaDO2, and that BALF-ADA activity may be a useful indicator of disease activity and the need for treatment.     

Predictive value of bronchoalveolar lavage in pulmonary sarcoidosis

We investigated whether analysis of cellular composition (including lymphocyte subsets) in bronchoalveolar lavage (BAL) fluid at the start of follow-up in patients with untreated sarcoidosis has any predictive value for further evolution of the disease. The outcome was evaluated by the chest roentgenograms, the lung volumes, and the single breath diffusing capacity for CO (DCO) after 22 to 36 months. In contrast to the general belief, patients who improved radiologically had a significantly higher T4 cell count (as percentage of BAL lymphocytes) (p less than 0.02) and a higher T4-T8 ratio in the initial BAL sample (9.3 vs 3.2; p less than 0.05) than those whose chest roentgenogram showed deterioration or remained unchanged. Total cell count and the percentage of lymphocytes in BAL fluid were not different between both groups. The change in DCO at the end of the follow-up period correlated positively with the baseline BAL T4 cells (Rs = 0.44; p less than 0.05) and with the BAL T4-T8 ratio (Rs = 0.51; p less than 0.03) and negatively with the baseline BAL T8 cells (Rs = -0.48; p less than 0.04). In only three patients progression of the disease necessitated steroid therapy, and they all had a low to normal T4-T8 ratio in the initial BAL sample. Bronchoalveolar lavage was repeated at least once in ten patients. Improvement of the chest roentgenograms in these patients was accompanied by a decrease of the BAL T4 cell count (as percentage of lymphocytes) and of the T4-T8 ratio. We conclude that a high lymphocyte count, a high T4 cell count (as percentage of lymphocytes), and a high T4-T8 ratio in BAL fluid reflect an intense alveolitis at the time of the procedure, but they are not indicators of poor prognosis on which therapeutic decisions can be based.     

Oxalic acid level in bronchoalveolar lavage fluid from patients with invasive pulmonary aspergillosis

Oxalic acid is a fermentation product of Aspergillus. We have measured the oxalic acid level in bronchoalveolar lavage fluids recovered from immunocompromised patients with and without invasive pulmonary aspergillosis. These levels were significantly higher in patients with invasive aspergillosis than in patients with pneumonitis of other causes. Thus, the determination of oxalic acid in bronchoalveolar lavage could be a presumptive argument for invasive aspergillosis until positive fungal cultures or histologic diagnosis; its potential value in monitoring the course of invasive pulmonary aspergillosis, particularly under treatment, has to be confirmed in more patients.     

Nitric oxide derivative in bronchoalveolar lavage fluid from patients with idiopathic pulmonary fibrosis

Idiopathic pulmonary fibrosis is a chronic inflammatory disorder of unknown aetiology with a number of inflammatory cells playing a role in its pathogenesis. In this study, we have attempted to find out the possible role of nitric oxide in its pathogenesis by way of measuring the nitrite and nitrate levels in the bronchoalveolar lavage fluid. Ten patients of histologically proved idiopathic pulmonary fibrosis and 10 controls (5 with normal chest skiagrams and 5 with sarcoidosis) were included in the study Bronchoalveolar lavage was carried out in these cases. The levels of nitrates and nitrites were increased in cases of idiopathic pulmonary fibrosis (0.77+/- 0.36 and 8.93 +/- 2.63 nmol/mg of protein) compared to those in controls (0.38 +/- 0.06 and 3.80 +/- 1.11, respectively for sarcoidosis patients); (0.39 +/- 0.13 and 6.56 +/- 1.61 for subjects with normal chest skiagrams). These differences were statistically significant (p < 0.05 to 0.01). These findings suggest a possible role of nitric oxide in the pathogenesis of idiopathic pulmonary fibrosis.     

Studies of bronchoalveolar lavage cells and fluids in pulmonary sarcoidosis. II. Enhanced capacity of bronchoalveolar lavage fluids from patients with pulmonary sarcoidosis to induce cell movement in vitro

The ability to increase the motility of endothelial cells in vitro is a property common to most if not all angiogenesis-inducing factors. Because bronchoalveolar lavage (BAL) cells from patients with pulmonary sarcoidosis have an enhanced capacity to induce neovascularization, the BAL fluids from these patients were assessed for their effect on human and murine endothelial cells and fibroblasts obtained from a variety of tissue sources. A recently developed computer-assisted image analysis system was used to determine the extent and pattern of cell migration in a microwell screening assay. Data were obtained for BAL fluids from 10 patients with pulmonary sarcoidosis and from five normal volunteers. BAL supernatants from patients with active sarcoidosis showed an enhanced (2- to 8-fold) capacity to induce chemokinesis of both endothelial cells and fibroblasts, as measured by increased area of migration and polarized cell movement. There was a marked heterogeneity in the motility of cells from different organ origins, but enhanced cell movement was observed with both endothelial cells and fibroblasts. In contrast, BAL fluids from normal and sarcoid patients were similar in their effect on muscle cells and urothelial cells, whereas pericytes, which responded to BAL fluids from normal subjects or patients with nongranulomatous pulmonary disease, were inhibited by BAL fluids from patients with pulmonary sarcoidosis. The induction of endothelial cell movement in vitro induced by individual supernatants generally correlated with the capacity of BAL cells from these patients to induce angiogenesis in vivo.(ABSTRACT TRUNCATED AT 250 WORDS)     

Inflammatory cytokine levels in induced sputum and bronchoalveolar lavage fluid in pulmonary sarcoidosis

BACKGROUND: The immune inflammatory process in patients with sarcoidosis is not only compartmentalized within the alveolar walls, but also involves the bronchial airways. Analysis of induced sputum has been used as a non-invasive tool for investigating the airways and may reflect the endobronchial and parenchymal inflammation in patients with sarcoidosis. This present study was designed to measure the soluble pro-inflammatory cytokine levels interleukin-1 (IL-1), interleukin-6 (IL-6), tumuor necrosis factor-alpha (TNF-alpha) and percentage of macrophages expressing these cytokines in induced sputum and bronchoalveolar lavage (BAL) fluid in patients with pulmonary sarcoidosis. METHODS: Sputum induction and BAL was carried out in 27 patients with newly diagnosed sarcoidosis. Control group consisted of six patients with a normal chest radiograph (three patients with carcinoma esophagus and three patients with doubtful history of hemoptysis). Induced sputum was also obtained from 10 non-smoking, non-atopic healthy controls. RESULTS: Percentage of macrophages expressing pro-inflammatory cytokines and soluble cytokine levels in induced sputum were higher in patients with sarcoidosis compared to both groups of controls. There was good correlation between IL-6 and TNF-alpha levels (r = 0.49, 0.58 p < 0.05) and percentage of macrophages expressing all three cytokines (r = 0.56-0.71, p < 0.01) between induced sputum and BAL fluid. Mild positive correlation between cytokine levels in sputum and age was also noted (r = 0.33-0.38, p < 0.05). CONCLUSIONS: Induced sputum may reflect changes in cytokine milieu in BAL in sarcoidosis.     

支气管肺泡灌洗液细胞类型与特发性肺纤维化预后的关联性研究

目的 探讨临床指标、肺功能和BALF中细胞类型与特发性肺纤维化(IPF)患者预后的关系.方法 经临床诊断的43例IPF患者进行肺功能和支气管肺泡灌洗检查.采用Kaplan-Meier检验比较组间生存率,采用Cox比例风险回归方法评价各参数的死亡风险度.结果 IPF患者存在限制性通气功能障碍和弥散功能障碍,FVC占预计值%、肺总量占预计值%和DLCO占预计值%分别为(61±18)%、(54±13)%和(48±14)%.在平均随访30.7个月内,IPF患者诊断后的中位生存期为28.5个月.糖皮质激素和(或)细胞毒类药物治疗不能改变IPF患者的预后.单因素Cox比例风险回归分析结果表明,FVC占预计值%、肺总量占预计值%、DLCO占预计值%、中性粒细胞和嗜酸粒细胞比例是影响IPF患者预后的因素,多因素Cox比例风险回归分析结果表明仅DLCO占预计值%和中性粒细胞比例是影响IPF患者预后的主要因素.结论 IPF患者主要为老年男性,存在限制性通气功能障碍和弥散功能障碍.糖皮质激素和(或)细胞毒类药物不能改变IPF患者的预后.DLCO占预计值%和中性粒细胞比例是影响IPF患者预后的主要因素,其中DLCO占预计值%和IPF患者的预后呈负相关,中性粒细胞比例和IPF患者的预后呈正相关.     

Lactate dehydrogenase in bronchoalveolar lavage fluid of patients with active pulmonary tuberculosis

BACKGROUND: Serum lactate dehydrogenase (LDH) concentration is an indicator of tissue injury. It may be increased in a variety of interstitial diseases and in pulmonary tuberculosis (PTB). OBJECTIVE: To investigate the value of LDH levels in bronchoalveolar lavage fluid (BALF) for the diagnosis of active PTB and to assess its relationship with serum LDH levels. METHODS: The study was a prospective clinical study. It included 25 consecutive patients with documented active PTB and 20 healthy adults who underwent bronchoscopy with bronchoalveolar lavage (BAL). Both total serum and LDH levels were measured. RESULTS: BALF LDH level was increased in all patients with active PTB. The mean BALF LDH level was significantly higher in patients with PTB (198.84+/-88.31 mIU/ml) as compared to controls (14.01+/-8.69 mIU/ml) (p = 0.0001). There was a significant correlation between BALF LDH and serum LDH levels in patients with PTB (r = 0.55, p = 0.006). CONCLUSION: BALF LDH levels are not specific and may be increased in many diseases. A very low value (possibly less than 60 mIU/ml) may, on the other hand, be useful to exclude the diagnosis of active PTB.     

Cellular profiles in bronchoalveolar lavage fluid of HIV-infected patients with pulmonary symptoms: relation to diagnosis and prognosis

Bronchoalveolar lavage (BAL) cell differentials and T-lymphocyte subpopulations were analysed in 95 HIV-infected patients with pulmonary symptoms to determine whether the type of cellular inflammatory response could be useful in diagnosis or as a prognostic marker. Patients with Pneumocystis carinii pneumonia (PCP) had more BAL fluid lymphocytes, mainly comprising CD8+ cells, and patients with bacterial infection had more neutrophils than other patients. Neither of these changes were mirrored in peripheral blood. Seven patients who died after their acute episode of PCP had significantly higher BAL fluid neutrophils than 53 patients with PCP who survived (P = 0.002). There seems to be correlation between BAL fluid neutrophilia, PCP and concomitant bacterial infection since four out of seven patients with a fatal outcome had coinfection with bacteria, whereas only one patient with PCP and bacterial coinfection survived (P = 0.0007).