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.     

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.     

Elevated levels of soluble TNF receptors in bronchoalveolar lavage fluid in extrinsic allergic alveolitis

BACKGROUND: Soluble TNF receptors (sTNFR1 and sTNFR2) are inhibitors of TNF and can block TNF bioactivity. TNF plays an important role in the development of extrinsic allergic alveolitis (EAA). OBJECTIVE: To evaluate whether sTNFR1 and sTNFR2 are locally increased in EAA. METHODS: We measured sTNFR1 and sTNFR2 in bronchoalveolar lavage fluid (BALF) and serum from nine EAA patients and 11 control subjects using an ELISA method. RESULTS: BALF sTNFR1 and sTNFR2 levels were 0.24+/- 0.04 ng/mL and 0.59+/-0.16 ng/mL in EAA patients, and thus significantly elevated in comparison with the controls (0.13+/-0.02 ng/mL and 0.08+/-0.04 ng/mL, both P<0.05). Serum sTNFR levels were not significantly different between the two groups. Both sTNFR1 and sTNFR2 concentrations in BALF correlated significantly with the lymphocyte percentage of BALF (r = 0.57 and 0.81, respectively). CONCLUSION: The two alveolar sTNFRs, particularly sTNFR2, may be involved in the pathogenesis of EAA as counter-regulators of TNF.     

CC chemokine receptor 4 ligand production by bronchoalveolar lavage fluid cells in cigarette-smoke-associated acute eosinophilic pneumonia

We examined the production of macrophage-derived chemokine (MDC/CCL22) and thymus- and activation-regulated chemokine (TARC/CCL17) by bronchoalveolar lavage fluid (BALF) cells in cigarette-smoke-associated acute eosinophilic pneumonia (CS-AEP). The CC Chemokine Receptor 4 (CCR4) ligand levels in BALF from patients with CS-AEP were considerably higher than those in healthy volunteers and correlated well with Th2 cytokine levels. Interleukin-4 enhanced CCR4 ligand production. MDC expression was observed in CD68-positive cells from patients with CS-AEP and in healthy control smokers. In contrast, TARC expression in CD68- or CD1a-positive cells was detected only in CS-AEP. An in vivo cigarette smoke challenge test induced increases in CCR4 ligands in the BALF and in the cultured supernatant of BALF adherent cells. These results suggest that alveolar macrophages and dendritic cells contribute to the pathogenesis of CS-AEP by generating CCR4 ligands, probably in response to cigarette smoke.     

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.     

Phospholipase activities of bronchoalveolar lavage fluid in rat Pneumocystis carinii pneumonia

Respiratory distress, main clinical symptom of P. carinii pneumonia, is unexplained. We wondered if it could be linked with a lung surfactant anomaly. We have shown in the bronchoalveolar fluid lavage of P. carinii-infected rats: a very significant decrease of the phospholipid content; a lowering of the phospholipid/protein ratio; and an increase in phospholipase activities, partly due to the activation of an inactive phospholipase into the active form. We conclude that, in P. carinii-infected rats, there is an increased catabolism of lung surfactant.     

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.     

Phospholipase activities of bronchoalveolar lavage fluid in rat Pneumocystis carinii pneumonia.

Respiratory distress, main clinical symptom of P. carinii pneumonia, is unexplained. We wondered if it could be linked with a lung surfactant anomaly. We have shown in the bronchoalveolar fluid lavage of P. carinii-infected rats: a very significant decrease of the phospholipid content; a lowering of the phospholipid/protein ratio; and an increase in phospholipase activities, partly due to the activation of an inactive phospholipase into the active form. We conclude that, in P. carinii-infected rats, there is an increased catabolism of lung surfactant.     

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.     

Accumulation of CXCR3-expressing eosinophils and increased concentration of its ligands (IP10 and Mig) in bronchoalveolar lavage fluid of patients with chronic eosinophilic pneumonia

BACKGROUND: Since human peripheral eosinophils have been shown to migrate to the CXC chemokine receptor 3 (CXCR3) ligands IFN-gamma-inducible protein 10 (IP10) and monokine induced by IFN-gamma (Mig), this confirms that CXCR3 is functionally expressed on these cells. IP10 expression has been shown to be increased in the airways of asthmatics. Eosinophil accumulations are found in bronchoalveolar lavage fluid (BALF) from patients with chronic eosinophilic pneumonia (CEP). To examine the contribution of IP10 and Mig in the pathogenesis of CEP, we measured the concentration of IP10 and Mig, and evaluated the expression of CXCR3 on eosinophils in BALF taken from patients with CEP. METHODS: The concentrations of IP10 and Mig in BALF were measured by ELISA. The proportion of CXCR3-expressing CD4+ T cells and CD16-negative eosinophils was determined by flow cytometry. RESULTS: The BALF concentrations of IP10 and Mig were higher in patients with CEP, as well as in patients with sarcoidosis, when compared to healthy controls. The absolute number of CXCR3+ CD4+ T cells was significantly higher in the BALF of patients with sarcoidosis, but not in the patients with CEP, when compared to healthy volunteers. There were higher percentages of CXCR3-expressing eosinophils in the BALF than in the peripheral blood of patients with CEP. CONCLUSIONS: Our findings suggest that IP10 and Mig contribute to the accumulation of CXCR3-expressing eosinophils in the lungs of patients with CEP, and modulate the eosinophilic inflammation of the lung.     

Cytologic examination of bronchoalveolar lavage fluid from immunosuppressed patients

Pulmonary infiltrates in immunocompromised patients frequently represent infections. This study was undertaken to evaluate cytologic examination of bronchoalveolar lavage samples and the value of different preparations and silver staining. Over 6 mo, 336 samples were collected from 155 immunosuppressed patients in whom both cytologic and microbiologic studies were performed. In 27 samples, the cytology, microbiology or both demonstrated the presence of infectious agents. In four cases, cytology identified neoplastic process. Cytology had a sensitivity of 34.6% for detection of infection, which increased to 42.9% when Cytomegalovirus was excluded. Cytology detected 6 of 15 cases of Aspergillus, including three cases not detected by microbiology and 3 of 4 cases of Pneumocystis, but did not identify any of the Cytomegalovirus cases. The type of preparations did not affect detection of the organism when present in cytologic samples and silver staining did not appear to have added value in examination of these samples.     

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.     

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 role of fibronectin in bronchoalveolar lavage fluid of asthmatic patients

Allergic and chronic inflammation of the airway is regarded as the main pathogenesis of bronchial asthma, in which adhesion of inflammatory cells requires the expression of adhesion molecules. Thus, to clarify the role of fibronectin (FN) in the airway inflammation of bronchial asthma, FN levels in plasma and bronchoalveolar lavage fluid (BALF) from bronchial asthmatics were determined. FN concentrations in plasma and BALF were measured by enzyme-linked immunosorvent assay (ELISA) in 17 asthmatic patients and 10 healthy controls to elucidate the role of FN in allergic inflammation. The mean FN/albumin (Alb) level in the BALF of asthmatic patients was 2.973 micrograms/mg, which was significantly higher than that of healthy controls (0.727 microgram/mg). Non-atopic asthmatics showed a significantly higher level of FN in their BALF in comparison with atopic asthmatics, although the ratio of FN to albumin showed no significant difference. FN levels in BALF correlated significantly with total cell density (r = 0.71, P < 0.05) and alveolar macrophage density (r = 0.64, P < 0.05). FN levels in plasma did not correlate with those in BALF. In conclusion, increased FN in BALF, which was produced locally in the airways of asthmatic patients, is actively involved in the regulation of allergic inflammation.