Accumulation of CCR4-expressing CD4+ T cells and high concentration of its ligands (TARC and MDC) in bronchoalveolar lavage fluid of patients with eosinophilic pneumonia

BACKGROUND: Th2 cells are thought to be involved in eosinophilic inflammation of the lung. CC chemokine receptor 4 (CCR4) has been identified as a specific receptor for both thymus and activation-regulated chemokine (TARC) and macrophage-derived chemokine (MDC), and is preferentially expressed on Th2 cells. OBJECTIVE: Our aim was to evaluate the role of Th2 cells in the lung of patients with eosinophilic pneumonia (EP). METHODS: The concentrations of TARC, MDC, and interleukin (IL)-5 were measured in bronchoalveolar lavage fluid (BALF) by ELISA. Proportion of CCR4-expressing CD4+ T cells (CCR4+ CD4+ T cells) was determined by flow cytometry. RESULTS: TARC and MDC concentrations in BALF were higher in patients with EP than in normal subjects. The proportion of CCR4-expressing cells among CD4+ T cells was higher in BALF than in peripheral blood of patients with EP. There was a significant correlation between the number of CCR4+ CD4+ T cells and the levels of TARC, MDC, and IL-5 in BALF of patients with EP. CONCLUSIONS: Our data suggest that Th2 cells, which express CCR4 and its ligands (TARC and MDC), contribute to the pathogenesis of EP in the lung.     

Elevated chemokine levels in bronchoalveolar lavage fluid of patients with eosinophilic pneumonia

BACKGROUND: Allergic lung inflammation is caused by accumulation and activation of different leukocyte subsets, such as eosinophils and T lymphocytes, in the lung. The chemokines are a large group of chemotactic cytokines that regulate leukocyte trafficking and may play an important role in allergic lung inflammation. OBJECTIVE: The purpose of this study was to evaluate the role of various chemokines, including eotaxin, RANTES, monocyte chemotactic protein (MCP)-1, macrophage inflammatory protein (MIP)-1beta, and IL-8 in the pathogenesis of eosinophilic pneumonia (EP). METHODS: The concentrations of eotaxin, RANTES, MCP-1, MIP-1beta, and IL-8 in bronchoalveolar lavage fluid (BALF) were measured by using ELISA in 15 patients with EP, 10 with idiopathic pulmonary fibrosis, 10 with sarcoidosis, and 11 healthy volunteers. RESULTS: Eotaxin in BALF was high only in patients with EP, and its level correlated significantly with the number of eosinophils in BALF of patients with EP and healthy volunteers. MCP-1 and MIP-1beta in BALF were preferentially increased in patients with EP. There was a significant correlation between MCP-1 levels and the number of macrophages in BALF of patients with EP and healthy volunteers. CONCLUSION: Our findings suggest that these CC chemokines contribute to the pathogenesis of EP through the specific recruitment of leukocyte subsets in the lung.     

Elevated interleukin-18 levels in bronchoalveolar lavage fluid of patients with eosinophilic pneumonia

BACKGROUND: Interleukin (IL)-18 can induce Th2 cytokine production particularly in collaboration with IL-2. Accumulation of Th2 cells and increased levels of Th2 cytokines are found in bronchoalveolar lavage fluid (BALF) from patients with eosinophilic pneumonia (EP). To evaluate the role of IL-18 in the pathogenesis of EP, we measured the concentration of IL-2, IL-12, IL-18, and Th2 cytokines in BALF from patients with EP. METHODS: The concentrations of interferon (IFN)-gamma, IL-2, IL-5, IL-10, IL-12, IL-13, and IL-18 in BALF were measured in patients with idiopathic acute eosinophilic pneumonia (AEP), with idiopathic chronic eosinophilic pneumonia (CEP), with sarcoidosis and healthy volunteers (HV). RESULTS: The BALF concentrations of Th2 cytokines, IL-5, IL-10, and IL-13, were higher in patients with EP than in sarcoidosis and control. The IL-2 level in BALF was higher in EP than in sarcoidosis and control. The IL-18 and IL-12 (p40 + p70) levels were higher in patients with EP than sarcoidosis, while the level of IL-12 (p70) was below the detection limit in patients with EP. There was a significant correlation between IL-2 level and both IL-5 and IL-13 in BALF of patients with EP. CONCLUSIONS: Our findings suggest that IL-18 may contribute to Th2 cytokine-dominant responses in patients with EP in collaboration with IL-2.     

Elevated soluble ADAM8 in bronchoalveolar lavage fluid in patients with eosinophilic pneumonia

BACKGROUND: ADAM (a disintegrin and metalloprotease) family members, characterized by a metalloprotease and a disintegrin domain, are membrane-anchored glycoproteins involved in proteolysis and cell adhesion. ADAM8 might have an important role in allergic inflammation. It can cleave a variety of substrates and is a sheddase for VCAM-1 and CD23, the low-affinity IgE receptors. METHODS: To evaluate the contribution of ADAM8 to the pathogenesis of eosinophilic pneumonia (EP), we measured the concentrations of soluble ADAM8 (sADAM8) and its substrates, soluble VCAM-1 (sVCAM-1) and soluble CD23 (sCD23), in bronchoalveolar lavage fluid from patients with smoking-induced acute eosinophilic pneumonia (AEP), chronic idiopathic eosinophilic pneumonia (CEP), and drug-induced eosinophilic pneumonia (drug-EP). RESULTS: The sADAM8 and sVCAM-1 concentrations were increased in AEP and CEP. The sCD23 concentration was elevated in AEP. In AEP, but not CEP, the sADAM8 concentration significantly correlated with those of both sVCAM and sCD23. CONCLUSION: The pathogenesis of AEP, CEP, and drug-EP was distinct with regard to ADAM8. Our results are the first to associate ADAM8 with eosinophilic responses and lung inflammation in humans.     

Overexpression of CD44 on alveolar eosinophils with high concentrations of soluble CD44 in bronchoalveolar lavage fluid in patients with eosinophilic pneumonia

BACKGROUND: High levels of interleukin-5 (IL-5) are found in the alveolar space of patients with eosinophilic pneumonia (EP). IL-5 promotes the growth and differentiation of eosinophils, as well as activating these cells. IL-5 also induces the expression of CD44 on eosinophils in vitro. To evaluate the contribution of CD44 to the pathogenesis of EP, we examined the expression of CD44 on eosinophils in bronchoalveolar lavage fluid (BALF) and measured the concentration of soluble CD44 (sCD44) in BALF from patients with EP. METHODS: The concentrations of IL-5, sCD44, and hyaluronic acid (HA) were measured in BALF. The expression levels of CD44 on eosinophils in BALF and peripheral blood in patients with EP were compared. RESULTS: The expression of CD44 on alveolar eosinophils and the concentration of sCD44 were increased in BALF of patients with EP. There was a significant correlation between IL-5 and sCD44 in BALF. A high concentration of HA was observed in BALF of EP patients. CONCLUSIONS: Our results suggest that the high expression of CD44 on eosinophils probably results from upregulation by IL-5 and could be important in the pathogenesis of EP.     

Detection of IL-5 and IL-1 receptor antagonist in bronchoalveolar lavage fluid in acute eosinophilic pneumonia

BACKGROUND: Acute eosinophilic pneumonia is an idiopathic cause of respiratory failure, characterized by very high numbers of alveolar eosinophils without significant blood eosinophilia. OBJECTIVE: The purpose of this study was to determine which cytokines are associated with acute eosinophilic pneumonia. METHODS: Soluble IL-1 type II receptor and the cytokines IL-1β, IL-1ra, IL-3, IL-5, granulocyte-macrophage colony-stimulating factor, and tumor necrosis factor-α were measured in serum and in bronchoalveolar lavage fluid from two patients with acute eosinophilic pneumonia during both acute and convalescent phases. RESULTS: Compared with patients with adult respiratory distress syndrome, the patients with acute eosinophilic pneumonia had high bronchoalveolar lavage fluid levels of IL-5, IL-1ra, and soluble type II IL-1 receptor but not IL-1β, tumor necrosis factor-α>, IL-3, or granulocyte-macrophage colony-stimulating factor. Bronchoalveolar lavage fluid levels of IL-5 and IL-1ra fell after resolution of symptoms. In the serum of patients with acute eosinophilic pneumonia, IL-5 was not detectable, and IL-1ra was initially high but fell after corticosteroid treatment. CONCLUSION: Acute eosinophilic pneumonia is characterized by locally high levels of IL-5, IL-1ra, and soluble type II IL-1 receptor in the alveolar space. (J ALLERGY CLIN IMMUNOL 1996;97:1366-74.)     

122例重症肺炎患儿支气管肺泡灌洗液的病毒检测分析

目的 了解湖南长沙地区重症肺炎患儿中呼吸道病毒的感染状况.方法 2011年1月至2011年12月,收集重症肺炎儿童的肺泡灌洗液标本122份,采用巢氏或普通逆转录聚合酶链反应(nested-PCR或RT-PCR)对腺病毒(ADV)、博卡病毒(HBoV)及副流感病毒1、2和4(PIV1、2、3、4)、呼吸道合胞病毒(RSV)、鼻病毒(HRV)、流感病毒A、B(IFVA-B)、人偏肺病毒(HMPV)、冠状病毒HKU1及NL63(HCoV-HKU1、HCoV-NL63)进行检测.结果 122份重症肺炎患儿肺泡灌洗液标本中,60份病毒阳性(49.1％),其中ADV检出率最高(40.98％,50/122),其次为RSV (7.37％,9/122),HBoV(7.37％,9/122).2种病毒混合感染有21例,占35％ (21/60),40％(20/50)的ADV阳性存在混合感染.结论 2011年,腺病毒感染是长沙地区重症肺炎的重要病因,其中腺病毒的混合感染在重症肺炎中常见.     

Pentraxin 3 in bronchoalveolar lavage fluid and plasma in non-neutropenic patients with pulmonary aspergillosis

Objectives

Pentraxin 3 (PTX3) contributes to resistance to Aspergillus infections. This study aimed to evaluate the presence of PTX3 in bronchoalveolar lavage fluid (BALF) and plasma in non-neutropenic patients with pulmonary aspergillosis.

Role of bronchoalveolar lavage in mechanically ventilated patients with suspected pneumonia

To determine the usefulness of samples obtained by bronchoalveolar lavage (BAL) in establishing the diagnosis of ventilator-associated pneumonia, quantitative cultures of BAL and protected specimen brush (PSB) samples obtained via fiberoptic bronchoscope were compared in 42 patients with suspected ventilator-associated pneumonia. Direct examination of BAL fluid was also used to identify cells with intracellular organisms. Ventilator-associated pneumonia was diagnosed in 18 patients; a total of 39 microorganisms were recovered from BAL fluid and 29 from PSB specimens. Cultures of 21 BAL and 23 PSB specimens were sterile. Quantitative BAL and PSB cultures coincided in 76 % of cases. Sterile BAL and PSB cultures agreed in 87 % of cases. Cultures were completely discordant in only three cases. The sensitivity of BAL for diagnosis of ventilator-associated pneumonia using bacterial counts of 10⁴ cfu/ml was 89 %, and specificity was 100 %. In 14 of the 18 patients with ventilator-associated pneumonia, the percentage of cells containing intracellular organisms in specimens recovered by BAL was 11.6 % versus 0.45 % in patients without pneumonia (p<0.05). In the remaining four patients, all of whom hadPseudomonas aeruginosa pneumonia, no intracellular organisms could be detected. Using a cut-off point of 5 % of cells with intracellular organisms, the sensitivity and specificity for the early diagnosis of ventilator-associated pneumonia was 67 % and 96 %, respectively. The results confirm the usefulness of the quantitative BAL culture (with a cut-off at 10⁴ cfu/ml) for the diagnosis of ventilator-associated pneumonia. The identification of intracellular organisms in BAL fluid is a good early indicator of pneumonia, but the sensitivity of this technique may be lower forPseudomonas aeruginosa infections.     

Detection and quantification of human herpesvirus 6 genomes using bronchoalveolar lavage fluid in immunocompromised patients with interstitial pneumonia

Human herpesviruses have been recognized as a pathogen involved in interstitial pneumonia (IP), especially in immunocompromised patients. So far, little is known about involvement of human herpesvirus 6 (HHV-6) in systemic respiratory tract disease. Currently, routine diagnostic tests for HHV-6 are inefficient for HHV-6 reactivation, therefore, we established a rapid quantification system of HHV-6 using real-time PCR in order to determine the possible role of human HHV-6 reactivation in immunocompromised patients showing IP. Bronchoalveolar lavage fluid (BALF) specimens were obtained from 84 consecutively treated patients with interstitial lung diseases (ILD) including various types of IP. First, we determined the viral burden in BALF and peripheral blood obtained from healthy volunteers. In healthy volunteers, the prevalence of HHV-6 in BALF was higher (4/12, 33.3%) than in peripheral blood (8/53, 15.1%), ranging from 0 to 101.65 HHV-6 genome copies per 1 microg of DNA. Among 84 patients with ILD analyzed, the prevalence of HHV-6 in BALF was 27.4% (23/84), ranging from 0 to 103.87 copies per 1 microg of DNA. Three specimens obtained from patients with collagen vascular disease, 2 from Hodgkin's disease, and 1 with sarcoidosis had high level of HHV-6 viral DNA, while none of the patients with idiopathic IP showed elevation of HHV-6 (more than 102) in BALF. Our results suggest that measurement of HHV-6 genomes in BALF using real-time PCR may be useful in management of the care of respiratory complications in immunocompromised patients.     

Eosinophilia in bronchoalveolar lavage fluid and architectural destruction are features of desquamative interstitial pneumonia

Aims:  Desquamative interstitial pneumonia (DIP) is a rare pattern of diffuse parenchymal lung disease known to overlap with respiratory bronchiolitis-interstitial lung disease (RB-ILD). The aim was to review biopsy-proven cases of DIP to investigate further the clinical, imaging and histological features of this disease.
Methods and results:  Twenty patients fulfilled the pathological criteria: 19 men and one woman with a mean age of 54 years. Clinical features, bronchoalveolar lavage (BAL) data, radiological findings, pathological findings other than criteria, effect of therapy and outcome were examined. The BAL data for 17 cases revealed marked eosinophilia (mean 18%) and moderate neutrophilia (mean 11%). Computed tomography in 17 patients showed peripheral involvement in all cases with a clear margin in 64% and thin-walled cysts in 35% of cases. Additional pathological features were a distinct lobular distribution (70%) and architectural destruction (70%) with cyst formation (55%). Eighteen of the 19 patients (95%) improved under steroid pulse and/or oral therapy. Sixteen subjects (80%) are alive, three died of other diseases and one died of DIP 74 months after the diagnosis. Percent vital capacity increased significantly and new thin-walled cysts appeared in one case.
Conclusions:  BAL eosinophilia, lobular distribution and architectural destruction with cyst formation are characteristic features of DIP.     

Chemokine and chemokine receptor analysis reveals elevated interferon-inducible protein-10 (IP)-10/CXCL10 levels and increased number of CCR5+ and CXCR3+ CD4 T cells in synovial fluid of patients with enthesitis-related arthritis (ERA)

Chemokines and chemokine receptors play a major role in homing of cells to the site of inflammation. Enthesitis-related arthritis (ERA) is a chronic inflammatory arthritis and no data are available on chemokines and their receptors in ERA. Blood (20) and synovial fluid (SF) (11) was collected from patients with ERA, and peripheral blood (PB) was collected from 12 patients with polyarticular juvenile idiopathic arthritis (JIA), nine patients with systemic onset and 18 healthy controls. Chemokines [interleukin (IL)-10/CXCL10, thymus and activation-regulated chemokine (TARC)/CCL17 and regulated upon activation normal T cell expressed and secreted (RANTES)/CCL5] were measured in serum and SF. Chemokine receptor expression was measured by flow cytometry. There was no difference in blood CD4(+) T cells bearing CCR5, CCR4 and CXCR3 in ERA and healthy controls. In paired samples the median frequency of CCR5(+) CD4(+) T cells was higher in SF compared to PB (15.8 versus 3.9%, P < 0.005), as was the frequency of CXCR3(+) T cells (21.61% versus 12.46%, P < 0.05). Median serum interferon-inducible protein-10 (IP-10)/CXCL10 levels were higher in patients with ERA compared to controls (139 versus 93 pg/ml; P < 0.05). Further median SF IP-10/CXCL10 levels were higher than the serum levels (2300 pg/ml versus 139 pg/ml; P < 0.01). Serum levels of RANTES/CCL5 were higher in patients (150 ng/ml) compared to control (99 ng/ml; P < 0.01). The SF levels were significantly lower compared to serum (P < 0.05). TARC/CCL17 levels in SF were lower than serum. There is increased homing of CCR5 and CXCR3(+) CD4 cells to the SF. Increased SF levels of IP-10/CXCL10 may be responsible for this migration in patients with ERA.     

Segmental allergen challenge in patients with atopic asthma leads to increased IL-9 expression in bronchoalveolar lavage fluid lymphocytes

BACKGROUND: IL-9 is a T(H)2 cell-derived cytokine that might be involved in the pathophysiology of allergic diseases. Little is known about its expression and release during the allergic response in the human lung. OBJECTIVE: The expression of IL-9 was measured in 10 atopic subjects with mild asthma and 5 nonatopic healthy control subjects at baseline and 24 hours after segmental sham and allergen challenge. METHODS: IL-9 protein was measured in bronchoalveolar lavage (BAL) fluid by means of ELISA and detected within the BAL cells by means of immunocytochemistry. Furthermore, IL9 mRNA expression of BAL cells was detected by means of real-time PCR. RESULTS: Although only low or undetectable amounts of IL9 mRNA and IL-9 protein were present in nonatopic control subjects and atopic asthmatic patients at baseline, there was an increase after segmental allergen challenge in the atopic subjects. Lymphocytes were identified as major cellular sources of IL-9 production by means of immunocytochemistry. Furthermore, IL-9 protein and IL9 mRNA expression correlated with eosinophil numbers in BAL fluid. CONCLUSIONS: These findings demonstrate that IL-9 is specifically upregulated after local allergen challenge in the lungs of atopic asthmatic patients. Lymphocytes are the major cellular source of IL-9. The increased expression and its correlation with eosinophil numbers suggest a potential role for IL-9 in the late phase of the allergic response.     

Glycosaminoglycans and glycoproteins in bronchoalveolar lavage fluid from patients with pulmonary alveolar proteinosis

Bronchoalveolar lavage fluid from two cases of pulmonary alveolar proteinosis were analyzed for glycosaminoglycans and glycoproteins. The clinical courses of the two cases were entirely different. In one patient, signs and symptoms recurred despite repeated therapeutic bronchoalveolar lavages. In the other patient, three successive bronchoalveolar lavages brought about complete recovery. It was found that the bronchoalveolar lavage fluid from the former case contained various subtypes of glycosaminoglycans [hyaluronic acid, chondroitin sulfate A(C), dermatan sulfate, and heparan sulfate] and glycoprotein. On the other hand, the bronchoalveolar lavage fluid from the latter case contained glycoprotein, but no detectable amounts of glycosaminoglycans. There was only a slight qualitative difference in glycoprotein of bronchoalveolar lavage fluid between the two cases. The presence or absence of glycosaminoglycans in bronchoalveolar lavage fluid may be related to the prognosis of pulmonary alveolar proteinosis.     

The chemokine receptor CXCR3 mediates rapid and shear-resistant adhesion-induction of effector T lymphocytes by the chemokines IP10 and Mig

Integrin-mediated adhesion to the vascular endothelium is an essential step in leukocyte diapedesis. We show that the chemokines 10-kDa inflammatory protein (IP10) and monokine induced by IFN (Mig) induce rapid and transient adhesion of human IL-2-stimulated T lymphocytes (IL-2 T cells) to immobilized integrin ligands through their receptor CXCR3, which is selectively expressed on activated T cells. Induction of adhesion by IP10 and Mig was already observed at subnanomolar concentrations and was maximal at 5 – 10 nM, resulting in three- to sixfold increase in adhesion of IL-2 T cells over background. No effect was seen with resting naive/memory T cells which lack CXCR3 and migration responses to IP10 and Mig. Both chemokines are produced in human umbilical vein endothelial cells (HUVEC) upon stimulation with IFN-γ and TNF-α. These chemokines induce IL-2 T cell adhesion also when captured on the surface of endothelial cells. Under conditions of flow, IL-2 T cells roll and rapidly adhere to IP10/Mig-expressing HUVEC, and anti-CXCR3 mAb treatment reduces arrest and firm adhesion. This is the first study that shows chemokine-induced adhesion in activated memory/effector T cells which represent the fraction of T cells that are selectively mobilized in inflammation. The critical role of IFN-γ as inducer of IP10/Mig production in HUVEC indicates that these chemokines are essential mediators of effector T cell recruitment to IFN-γ-dependent pathologies.     

IgE-Fc receptor expressions on bronchoalveolar lavage (BAL) cells in a patient with eosinophilic pneumonia accompanied with bronchial asthma--the effect of ketotifen on their expression

In this study, IgE-Fc receptor expressions on each type of BAL cells (lymphocytes, macrophages and eosinophils) in a patients with eosinophilic pneumonia accompanied by bronchial asthma were examined by indirect immunofluorescent method using monoclonal antibody (H107). BAL cell findings showed marked increases of total BAL cell counts and eosinophils, an increased number of lymphocytes and the presence of basophilic cells. These results match those of our previous report. Furthermore, IgE-Fc receptor expressions on lymphocytes, macrophages and eosinophils were markedly increased as compared to those in the peripheral blood and normal control subjects. These findings suggest that IgE production in the lung plays the main role in the pathogenesis of eosinophilic pneumonia and bronchial asthma. On the other hand, during ketotifen administration, decreases of peripheral blood eosinophilia, of eosinophilia in the sputum and of serum IgE level and an improvement in chest X-ray findings were observed. Furthermore not only normalization of total BAL cell counts but also decreases of IgE-Fc receptor expressions on BAL cells were observed.     

Eosinophilic leucocytes and arylsulfatase activity in bronchoalveolar lavage fluid of patients with bronchial asthma

The arylsulfatase activity and histamine concentration of bronchoalveolar lavage fluid (BALF) were examined in patients with bronchial asthma in relation to the eosinophil count and asthma type (atopic and non-atopic). The BALF arylsulfatase activity and histamine concentration were significantly higher in atopic asthmatics than in non-atopic asthmatics. In atopic asthmatics, the activity of arylsulfatase was significantly increased in patients with a higher eosinophil count (10% or more). However, the BALF histamine concentration did not correlate with the eosinophil count. In non-atopic asthmatics, there was no significant correlation between arylsulfatase activity and the eosinophil count. The results show that arylsulfatase participates in IgE-mediated allergic reactions.     

Change in the concentration of neutrophil elastase in bronchoalveolar lavage fluid during anesthesia and its inhibition by cholesterol sulfate

Cholesterol sulfate (CS) in the gastrointestinal tract exhibits a mucosal protective activity in mouse ulcer model. To clarify the possible role of CS for protection from the epithelial injury due to neutrophil elastase in the tracheobronchi, the authors determined the concentrations of CS and neutrophil elastase in bronchoalveolar lavage fluid (BALF) from patients under anesthesia, and they examined the inhibitory activity of CS toward neutrophil elastase. The concentrations of CS and neutrophil elastase were determined by thin-layer chromatography and enzyme-linked immunosorbent assaying, respectively, and the effect of CS on the activity of elastase was determined with a chromogenic substrate. CS was found to be present in human lung, tracheal mucosa, and BALF, and a high synthesis of it was detected in the tracheal mucosa, in which cellular cholesterol sulfotransferase was induced depending on the density of tracheal cells. Among lipids in the tracheal mucosa, only CS was demonstrated to exhibit inhibitory activity toward neutrophil elastase, a powerful erosive agent in inflammation. The secretion of elastase from neutrophils into BALF was stimulated during the course of general anesthesia. In contrast, the amount of CS in BALF gradually decreased during anesthesia. On immune-precipitation of neutrophil elastase in BALF, CS was detected in the immune precipitate, which indicates a possible association of CS with neutrophil elastase in BALF. Conclusion: CS, which is a major acidic lipid in the tracheobronchial epithelium, might function as an epithelial inhibitor toward neutrophil elastase secreted in response to several stimuli such as anesthesia.     

Analyses of bronchoalveolar lavage fluid (BALF) in MRL-lpr/lpr mice

We performed bronchoalveolar lavage (BAL) in MRL-lpr/lpr (MRL/l) and MRL- +/+ (MRL/n) mice and evaluated various cellular and humoral components of the bronchoalveolar lavage fluid (BALF) to clarify the pathogenic mechanism of pulmonary fibrosis in MRL/l mouse. The numbers of macrophages, neutrophils and lymphocytes, N-Acetyl-beta-glucosaminidase (beta-NAG), and fibronectin increased in the BALF from MRL/l mice than that from MRL/n mice, but no significant differences were observed in total protein, beta-glucuronidase, acid phosphatase, or phospholipid level. Increased fibronectin level in the BALF from MRL/l mice may be related with pathogenesis of pulmonary fibrosis.