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PubMed2002

Matrix metalloproteases in BAL fluid of patients with cystic fibrosis and their modulation by treatment with dornase alpha.

BACKGROUND: Matrix metalloproteinases (MMPs) are involved in the remodelling and degradation of extracellular matrix and may play a role in pulmonary tissue destruction in cystic fibrosis (CF). METHODS: Bronchoalveolar lavage (BAL) fluid levels of MMP-8, MMP-9, and their natural inhibitor TIMP-1 were measured on two occasions within 18 months in 23 children with mild CF, 13 of whom were treated with DNase. RESULTS: MMP-8 (39.3 (6.8) v 0.12 (0.01) ng/ml), MMP-9 (58.0 (11.4) v 0.5 (0.02) ng/ml), and the molar ratio of MMP-9/TIMP-1 (0.36 (0.05) v 0.048 (0.01)) were significantly higher in patients with CF than in control children without lung disease. Gelatine zymography showed the typical banding pattern of neutrophil derived MMP-9, including 130 kDa NGAL-MMP-9 complex and 92 kDa latent MMP-9 bands; 85 kDa bands (corresponding to active MMP-9) were seen in all patients. There was a close correlation between BAL fluid concentrations of MMPs and alpha(2)-macroglobulin, a marker of alveolocapillary leakage. After 18 months MMP levels were increased in untreated patients and decreased in patients treated with DNase. CONCLUSIONS: Uninhibited MMPs may contribute to pulmonary tissue destruction even in CF patients with mild lung disease that may be positively affected by treatment with DNase.

Adolescent↗
PubMed1990

Corticosteroids increase lipocortin I in BAL fluid from normal individuals and patients with lung disease.

Lipocortin I is a corticosteroid-inducible protein that has potent anti-inflammatory activity. To determine whether lipocortin I is present on the epithelial surface of the human lung, we used a specific polyclonal antibody by the technique of Western blotting to evaluate bronchoalveolar lavage (BAL) fluid of normal individuals and patients with idiopathic pulmonary fibrosis. Lipocortin I was a normal constituent of the epithelial surface of the normal lung and comprised 0.23 +/- 0.03% of BAL fluid proteins. Four separate immunoreactive species were detected, at 37, 36, 34, and 33 kDa, consistent with previously published results. Corticosteroids increased the amounts of lipocortin present in normal volunteers and in patients with idiopathic pulmonary fibrosis. These results demonstrate that lipocortin I is normally present in the human lung and further suggest that lipocortin I may be an important modulator of the anti-inflammatory effects of corticosteroids in the lung.

Adrenal Cortex Hormones↗
PubMed1988

The intensity of sarcoidosis alveolitis is related to albumin and IgG levels in BAL.

To define whether the levels of immunoglobulins and albumin in bronchoalveolar lavage (BAL) fluid are related to the intensity of sarcoidosis alveolitis, 49 patients affected with pulmonary sarcoidosis were studied. To distinguish between high- and low-intensity alveolitis, two parameters were used: the percentage of T lymphocytes and the CD4+/CD8+ ratio in BAL fluid. The results showed an increase of albumin and of the IgG/albumin ratio (p less than 0.05) in patients with high-intensity alveolitis as assessed by both criteria. Moreover, an increase (p less than 0.05) of albumin concentration related to T lymphocyte alveolitis was found. We conclude that: albumin is a reliable index of inflammation; the increase of the IgG/albumin ratio is the expression of local production of IgG as well as of inflammation; the evaluation of albumin and of the IgG/albumin ratio should be useful because they both correlate with the intensity of the alveolitis: the former express inflammatory damage and the latter, immunological activity.

Adult↗
PubMed1992

Correlation between cell content of bronchoalveolar lavage (BAL) and histologic findings in asthma.

We have recently examined the relationship between cell content of bronchoalveolar lavage (BAL) fluid and lung histology in a group of 13 asthmatic subjects during clinical remission. Inflammatory cells in BAL appeared to reflect the severity of the overall inflammatory process of the bronchial mucosa. In the present report we examined whether in asthma the presence of a thickened basement membrane is associated with inflammation of the bronchial mucosa. There was a significantly higher number of inflammatory cells (p less than 0.01) and eosinophils (p less than 0.005) in the submucosa of subjects showing a thickened basement membrane (greater than 7 microns) compared to those having normal basement membrane (less than or equal to 7 microns). These findings seem to support the hypothesis that the degree of thickening of basement membrane in asthma is related to mucosal inflammation.

Adult↗
PubMedDate not supplied

Diagnosis of ventilator-associated pneumonia: focus on nonbronchoscopic techniques (nonbronchoscopic bronchoalveolar lavage, including mini-BAL, blinded protected specimen brush, and blinded bronchial sampling) and endotracheal aspirates.

The ideal diagnostic approach for ventilator-associated pneumonia currently is based on invasive procedures to obtain respiratory tract cultures. Given the lack of consensus and relatively poor acceptance of full bronchoscopic bronchoalveolar lavage (BAL) and protected specimen brush (PSB), less invasive procedures have been developed. We review the nonbronchoscopic procedures (nonbronchoscopic bronchoalveolar lavage, including mini-BAL, blinded protected specimen, and blinded bronchial sampling) and endotracheal aspiration. We provide a critique of the methods used, the types of catheters inserted, and the sample collection methods. Most studies were flawed in that antibiotic use before initiation of the procedure was not controlled. The variability of both the methods and the criteria for the gold standard in the numerous investigations show that these procedures are neither standardized nor proven to be accurate and often did not improve management. Pending future studies, use of endotracheal aspirates without the use of quantitation seems to be a reasonable approach for clinicians who are not committed to an invasive procedure.

Bronchi↗
PubMed2004

Predictive value of BAL cell differentials in the diagnosis of interstitial lung diseases.

The current authors aimed to quantify how the likelihood for a given diagnosis changes with the knowledge of bronchoalveolar lavage (BAL) cell differentials. As an initial estimate (a priori probability), frequencies of final diagnoses were taken. Using categorisations for cell differentials, a posteriori probabilities were then derived for each disease, according to Bayes. The analysis was performed in three of five groups of diagnoses suspected prior to BAL: interstitial lung disease (ILD; n=710), inflammatory disease (n=583), or lung tumour mimicking ILD (n=455). Overall, out of 1,971 patients, 18.3% had sarcoidosis, 7.7% usual interstitial pneumonia (UIP), 4.4% extrinsic allergic alveolitis (EAA), and 19.0% tumours. In the group with suspected ILD, the likelihood for sarcoidosis increased from 33.7 to 68.1% when lymphocyte numbers were 30-50% and granulocyte numbers were low; the likelihood for UIP increased from 15.8 to 33.3% when lymphocyte numbers were <30% with granulocytes elevated. CD4/CD8 was informative, especially in sarcoidosis and EAA. Despite considerable increases, the likelihood of rare diseases rarely reached appreciable values. Similar results were obtained in the other two groups of suspected diagnoses. In conclusion, these data suggest that bronchoalveolar lavage cell counts per se provide substantial diagnostic information only in relatively frequent diseases, such as sarcoidosis and usual interstitial pneumonia, and are less helpful in infrequent diseases.

Analysis of Variance↗
PubMed1996

Oxidized BAL fluid proteins in patients with interstitial lung diseases.

Oxygen-derived free radicals, released by phagocytic cells, have been postulated to contribute to lung tissue damage. We therefore investigated oxidative damage to proteins from bronchoalveolar lavage fluid (BALF) as an indicator of oxidative stress and to assess antioxidant defences in the lungs. We examined BAL fluids from patients with interstitial lung diseases, such as idiopathic pulmonary fibrosis (IPF, nonsmokers (NS) and smokers (S)), sarcoidosis (SARC, nonsmokers), and asbestosis (ASB, ex-smokers (EXS)). The oxidation of BALF proteins is accompanied by the introduction of carbonyl groups into their amino acid side-chains and can be quantitated by labeling these groups with tritiated borohydride. The total lung content of oxidized proteins recovered by bronchoalveolar lavage (BAL) was 0.3 +/- 0.07 nmol carbonyl.mL-1 BALF (mean +/- SEM) in the NS control group (n = 9) and tended to be increased, in the asymptomatic S group (n = 8; 0.59 +/- 0.14 nmol.mL-1). This parameter was significantly elevated both in IPF-NS (n = 14; 0.84 +/- 0.2 nmol carbonyl.mL-1 BALF) and SARC-NS (n = 15; 0.73 +/- 0.16 nmol.mL-1) as compared with the NS control. On the contrary, in smoking patients with IPF (n = 6; 0.41 +/- 0.1 nmol carbonyl.mL-1 BALF) and also in ASB-EXS (n = 6; 0.37 +/- 0.06 nmol.mL-1) it was not different from NS controls. The total amount of oxidized proteins correlated positively with the absolute number of eosinophils (EOS) in IPF-NS, IPF-S and SARC, and also with absolute polymorphonuclear neutrophil (PMN) numbers in IPF-NS and IPF-S. In conclusion, oxidative damage of BALF proteins occurred in nonsmoking patients with IPF and SARC. The amount of oxidized bronchoalveolar lavage fluid protein may provide a quantitative assessment of oxygen burden, a balance between oxidant stress and antioxidant defences.

Adult↗
PubMed1996

Integrin alpha E beta 7 expression on BAL CD4+, CD8+, and gamma delta T-cells in bleomycin-induced lung fibrosis in mouse.

CD4, CD8, and gamma delta T-cells located in the epithelium express the integrin alpha E beta 7 that binds to E-cadherin on the epithelium. Gamma delta T-cells mediate specific cellular immune functions and can recognize damaged cells directly. It was, therefore, of interest to analyse the presence of gamma delta T-cells and the expression of alpha E beta 7 on gamma delta T-cells in the bleomycin (BLM) model of pulmonary fibrosis. Lung fibrosis was induced by a single intratracheal instillation of BLM (0.125 U.mouse-1), and bronchoalveolar lavage (BAL) T-cell subpopulations were examined at various time-points for the expression of the integrin alpha E beta 7 by flow cytometry. CD4+ T-cells accounted for about 40% of the lymphocytes, compared to about 10% of CD8+ T-cells and 10-14% gamma delta T-cells. Within the CD4+ T-cell population the proportion of alpha E beta 7+ cells decreased between Days 2 and 22 from 36 to 11%. The percentage of alpha E beta 7+ CD8+ T-cells increased at the same time from 4 to 68%. However, more than 80% of the gamma delta T-cells in BAL fluid expressed alpha E beta 7 at all time-points. The surface-expression of this integrin on gamma delta T-cells was 2-3 times higher than on CD4+ or CD8+ T-cells. This predominant expression of alpha E beta 7 on gamma delta T-cells suggests a role for these cells in the pathogenesis of bleomycin-induced lung fibrosis.

Animals↗
PubMed1997

Serum and BAL Clara cell 10 kDa protein (CC10) levels and CC10-positive bronchiolar cells are decreased in smokers.

Cigarette smoking has diverse effects on the structure and function of the lung. Smoking appears to reduce the levels of Clara cell 10 kDa protein (CC10) in the alveolar lining fluid, but the influence of smoking serum on CC10 levels is still debated, and it has not been clear whether smoking reduces the number of CC10-producing lung cells. The aims of this study were to clarify the influence of smoking on CC10 levels in the alveolar lining fluid and bloodstream, and on the number of CC10-producing lung cells. CC10 concentrations were measured in sera and bronchoalveolar lavage (BAL) fluids, by means of enzyme-linked immunosorbent assay using monoclonal and polyclonal antibody, and the immunohistochemical expression of CC10 was examined in the lungs of nonsmokers and smokers using the monoclonal antibody, TY-5, against CC10/human urinary protein-1. CC10 concentrations in sera and in BAL fluids from healthy smokers were significantly lower than in healthy nonsmokers. Immunohistochemical expression of CC10 was found exclusively in nonciliated bronchiolar epithelial cells. As compared to that of nonsmokers, the mean percentage of CC10-positive bronchiolar epithelial cells was significantly decreased in lung tissue specimens obtained from smokers who had normal results in pulmonary function tests. It was concluded that smoking reduces the proportion of Clara cell 10 kDa protein-producing bronchiolar epithelial cells, resulting in decreased levels of Clara cell 10 kDa protein in the lower respiratory tract and in the bloodstream. The protein is a new blood biochemical and immunohistochemical marker, reflecting structural changes in peripheral airways induced by cigarette smoking.

Adult↗
PubMed1997

Airway versus transbronchial biopsy and BAL in lung transplant recipients: different but complementary.

Lung transplantation is now an established therapeutic intervention for end-stage cardiopulmonary disease in humans. Chronic rejection, in the form of bronchiolitis obliterans syndrome (BOS), remains the commonest cause of morbidity and mortality in those surviving more than 3 months. The pathology of BOS involves airway changes. We have evaluated the potential for endobronchial biopsies (EBB) to complement existing sampling methods used in allograft monitoring and have compared the results of EBB findings with those of bronchoalveolar lavage (BAL) and transbronchial biopsy (TBB) in 18 clinically stable patients. We found that all the EBB had inflammatory cells present but that only five TBB specimens had evidence of inflammation, with airway material being present in 78% of the TBB. Paired BAL and EBB yielded different results, with no correlations between total macrophages, lymphocytes, CD4+ cells or CD8+ cells. We conclude that endobronchial biopsies are potentially useful as an additional sample for the monitoring of inflammation in lung allografts, since they yield different, and potentially complimentary, information to bronchoalveolar lavage and transbronchial biopsy.

Adult↗
PubMed1996

Increased expression of platelet-derived growth factor A and insulin-like growth factor-I in BAL cells during the development of bleomycin-induced pulmonary fibrosis in mice.

Current concepts suggest that macrophages may play a central role in pulmonary fibrosis by virtue of their ability to release a variety of cytokines. In this study, the expression of interleukin (IL)-1 alpha and beta, platelet-derived growth factor (PDGF) A and B, and insulin-like growth factor (IGF) I in BAL cells, which may be involved in fibroblast proliferation, was investigated in murine bleomycin (BLM)-induced pulmonary fibrosis. BAL cells were obtained at 1, 15, and 29 days from Institute for Cancer Research mice after 10 days of intraperitoneal administration of BLM. The relative amounts of cytokine messenger RNA (mRNA) were evaluated by the reverse transcription-polymerase chain reaction method, which simultaneously amplified complementary DNA for cytokines and beta-actin as an internal control. The level of IL-1 beta mRNA in BLM-treated mice was increased 4.5-fold compared with that in saline solution-treated (control) mice 1 day after treatment, while no significant differences were observed between the two groups at 15 and 29 days. The mRNAs of PDGF-A and IGF-I in BLM-treated mice were sustained at levels eightfold and threefold to fourfold, respectively, those of controls over 4 weeks. No significant differences were noted in IL-1 alpha and PDGF-B expression between the two groups. We conclude that IL-1 beta released from macrophages may be important in the early phase of inflammatory responses and that PDGF-A and IGF-I may play important roles in the development of BLM-induced pulmonary fibrosis.

Animals↗
PubMed1996

Decreased contents of surfactant proteins A and D in BAL fluids of healthy smokers.

Hydrophilic surfactant proteins, surfactant protein A (SP-A) and surfactant protein D (SP-D), have important roles in modulating the host defense functions in the peripheral airways. It has been reported that cigarette smoke may alter the component and function of pulmonary surfactant. In this study, we determined the contents of SP-A and SP-D in BAL fluids of healthy smokers and nonsmokers by enzyme-linked immunosorbent assay using monoclonal antibodies against each protein. The contents of SP-A and SP-D in BAL fluids were significantly (p<0.05) decreased in smokers compared to those in nonsmokers, although there was no significant difference of total phospholipid content between smokers and nonsmokers. These results suggest that the decreased levels of SP-A and SP-D in smokers may impair the host defense functions of surfactant in the peripheral airways and might have a crucial roles in the development of chronic obstructive lung disease.

Adolescent↗
PubMed2000

Imbalances between tumor necrosis factor-alpha and its soluble receptor forms, and interleukin-1beta and interleukin-1 receptor antagonist in BAL fluid of cavitary pulmonary tuberculosis.

OBJECTIVES: We investigated the possibility that the large pulmonary cavity in tuberculosis (TB) lesions might result from imbalances between tumor necrosis factor-alpha (TNF-alpha) and soluble TNF-alpha receptor forms (sTNF-RI and sTNF-RII), and interleukin-beta (IL-1beta) and IL-1 receptor antagonist (IL-1RA) in sites of local inflammation. PATIENTS AND METHODS: BAL was performed in 32 patients with active pulmonary TB, and the recovered BAL fluid (BALF) was examined for concentrations of TNF-alpha and its soluble receptor forms, IL-1beta, and IL-1RA. Patients were classified into two groups: group 1, patients with a large cavity (> or = 4 cm) on chest radiographs (n = 15); and group 2, patients with a small cavity (< 4 cm; n = 3) or no cavity (n = 14) on chest radiographs. RESULTS: The concentrations of TNF-alpha, IL-1beta, and IL-1RA in BALF were significantly higher in group 1 patients than in group 2 patients before standardization. The difference was also statistically significant for TNF-alpha and IL-1beta after standardization with urea. Furthermore, group 1 patients had significantly higher ratios of TNF-alpha to sTNF-RI and sTNF-RII and IL-1beta to IL-1RA compared with group 2 patients. CONCLUSIONS: These findings suggest that the relative abundance of TNF-alpha and IL-1beta associated with imbalances of secretion of soluble TNF-alpha receptor forms and IL-1RA may have caused tissue necrosis leading to cavity formation in patients with active pulmonary TB.

Adult↗
PubMed2000

The role of BAL in the diagnosis of pulmonary mucormycosis.

Five patients with pulmonary mucormycosis diagnosed during life are described. All had underlying predisposing conditions: either posttransplant or hematologic malignancies. In all cases, the diagnosis was made using fiberoptic bronchoscopy. In three patients, BAL was diagnostic. In two of these patients, the diagnosis was made by identifying the typical hyphae of mucormycosis in the BAL fluid alone. Transbronchial biopsy was diagnostic in three patients. Treatment was based on IV antifungal chemotherapy together with surgical removal of involved lung tissue whenever feasible. The clinical outcome of these patients was dismal and was determined primarily by the underlying condition.

Adult↗
PubMed2002

Diagnosis of invasive pulmonary aspergillosis using polymerase chain reaction-based detection of aspergillus in BAL.

STUDY OBJECTIVE: To assess the value of Aspergillus polymerase chain reaction (PCR) test performed on the BAL in diagnosing invasive pulmonary aspergillosis (IPA). DESIGN: Between January 1996 and 1997, we prospectively followed up 249 cancer patients with pulmonary infiltrates suggestive of pneumonia. Bronchoscopy with fungal stains, cultures, and PCR was performed on all patients. PCR was used for the detection of Aspergillus mitochondrial and alkaline protease gene DNA. The PCR products were visualized either directly on polyacrylamide gel or after Southern transfer and probing with specific probes for mitochondrial and alkaline protease DNA. RESULTS: The 249 patients consisted of 10 patients with proven IPA (tissue invasion), 22 patients with probable IPA (microbiologic culture), 18 patients with possible IPA (consistent clinical and radiologic findings), and 199 control patients with no evidence of IPA. PCR positivity was strongly associated with all forms of IPA (p < 0.002). The sensitivity, specificity, positive predictive value, and negative predictive value of PCR were 80%, 93%, 38%, and 99%, respectively, for proven IPA, and 64%, 93%, 52%, and 96%, respectively, for probable IPA. Southern blotting analysis did not improve the diagnostic yield of the PCR test. CONCLUSION: PCR performed on BAL is associated with high specificity and negative predictive value for IPA. The low positive predictive value could be related to the transient colonizing presence of aspergilli in the respiratory tract. The sensitivity correlates with the certainty of the diagnosis based on tissue invasion.

Adolescent↗
PubMed1981

The metabolism of arsenic in humans acutely intoxicated by As2O3. Its significance for the duration of BAL therapy.

The urinary excretion of inorganic arsenic and of its less toxic metabolites [monomethylarsonic acid (MMA) and dimethylarsinic acid (DM)] has been followed-up in five cases of acute oral intoxication by As2O3. In addition to supportive therapy all patients were given BAL. The proportion of the three species of urinary arsenic (inorganic, MMA, DMA) changes markedly with time. During the first 2 to 4 d after the intoxication, arsenic is excreted mainly as its unmetabolized inorganic form but this is rapidly followed-up by a progressive increase of the proportion excreted as MMA and DMA. The time at which the majority of arsenic is excreted as its organic metabolites depends on the severity of the intoxication but in all the cases, more than 95% of th excreted arsenic is in the organic form after 9 d, the dimethylated derivative being preponderant. Speciation of arsenicals in urine might be useful to determine the duration of BAL treatment.

Adult↗
PubMedDate not supplied

Detection and qualitative identification of mineral fibers and particles in alveolar macrophages of BAL fluid by SEM and EDXA.

Inorganic dust inhalation diseases represent one of the most important chapters in respiratory medicine because of their diagnostic, therapeutic, legal, ecological and social implications. While, in fact, toxic substances inhalation may be easily related to particular occupations, it is more difficult to recognize the potential damage represented by occasional and fortuitous exposition due to pollution of one's living environment. The aim of this study was to suggest a useful investigative method for detecting the presence of mineral substances (dusts and fibers) in the lung in pulmonary fibrosis of uncertain origin. We used scanning electron microscopy (SEM) and semi-quantitative energy-dispersive x-ray microanalysis (EDXA) on broncholaveolar lavage (BAL) and sputum samples of 10 patients, all males, aged 41-66 years, smokers, affected by interstitial lung disease. Two subjects had a negative professional anamnesis while the other 8 declared a potential exposition to inorganic toxic dusts: 2 subjects were involved in the production of asbestos-containing building materials, 2 were miners, 1 a ceramic worker, and 3 insulating materials handlers. Data are reported on the detection of asbestos bodies, vitreous fibers and silica content of alveolar macrophages in BAL fluid.

Adult↗
PubMed1992

Value of CEA and SCC-Ag in bronchoalveolar lavage (BAL) and serum of patients with lung carcinoma.

To evaluate the diagnostic usefulness of simultaneous determinations of carcinoembryonic antigen (CEA) and squamous cell carcinoma antigen (SCC-Ag), we studied 25 patients with lung carcinoma and 12 with nonmalignant lung diseases. The measurements were made in serum and bronchoalveolar lavage (BAL). The results showed that positive rates with lavage CEA and SCC-Ag in lung carcinoma patients were higher in comparison with those markers in serum. The combination of lavage CEA and SCC-Ag taken together with the results of bronchoscopy (histology and cytology) showed the highest discriminating power between malignant and nonmalignant lung diseases. The sensitivity of bronchoscopy increased from 48 to 84% with at least 1 positive marker. It appears that the simultaneous determination of CEA and SCC-Ag levels in serum and BAL in lung carcinoma patients may be of considerable importance in diagnosis.

Adenocarcinoma↗