Bronchoalveolar Fluid Of Patients Research Articles

Overexpression of Human Aspartyl (Asparaginyl) β-hydroxylase in NSCLC: Its Diagnostic Value by Means of Exosomes of Bronchoalveolar Lavage.

The human aspartyl β-hydroxylase (ASPH) is overexpressed in tumor tissues. Bronchoalveolar lavage (BAL) is a diagnostic procedure for infections and malignancies. The aim of this study was to investigate whether tumor exosomes carrying ASPH gene marker were present in bronchoalveolar fluid of patients with non-small cell lung cancer (NSCLC). A tissue microarray analysis was applied to explore the expression of ASPH in different histologic NSCLC. The human NSCLC cell lines and normal bronchial cell lines were used to study exosomal ASPH exprerssion. A total of 27 NSCLC, 21 benign tumor, and 15 healthy controls underwent BAL. Immunohistochemistry was performed to study the ASPH expression in malignant and normal lung tissues. The expression characteristics of ASPH in different NSCLC and normal bronchial cells and pneumocytes were confirmed by cell blocks. A reverse transcription-quantitative polymerase chain reaction was carried out to study the levels of exosomal ASPH expression. Immunohistochemical staining of tissue microarray demonstrated that overexpression of ASPH was found in NSCLC tissues including adenocarcinoma, large cell carcinoma, and squamous cell carcinoma, but absent in adjacent normal tissues. All NSCLC specimens exhibited high levels of ASPH immunoreactivity, while nonmalignant and normal lung tissues exhibited a very low level of expression. Overexpression of ASPH was found in exosomes from NSCLC cell lines but absent from the normal bronchial cell line NL-20. ASPH level from BAL exosomes was significantly increased in NSCLC patients compared with that from nonmalignant or health group. Our method of isolation of BAL exosomes was easily performed in the clinical laboratory. BAL exosomal ASPH can be a potential biomarker for NSCLC diagnosis.

Monocytes and macrophages in severe COVID-19 - friend, foe or both?

Monocytes and macrophages in severe COVID-19 - friend, foe or both?

Role of endothelial microRNA 155 on capillary leakage in systemic inflammation

BackgroundCapillary leakage is a key contributor to the pathological host response to infections. The underlying mechanisms remain incompletely understood, and the role of microRNAs (MIR) has not been investigated in detail. We hypothesized that specific MIRs might be regulated directly in the endothelium thereby contributing to vascular leakage.MethodsSmallRNA sequencing of endotoxemic murine pulmonary endothelial cells (ECs) was done to detect regulated vascular MIRs. In vivo models: transgenic zebrafish (flk1:mCherry/l-fabp:eGFP-DPB), knockout/wildtype mouse (B6.Cg-Mir155tm1.1Rsky/J); disease models: LPS 17.5 mg/kgBW and cecal ligation and puncture (CLP); in vitro models: stimulated human umbilical vein EC (HUVECs), transendothelial electrical resistance.ResultsEndothelial MIR155 was identified as a promising candidate in endotoxemic murine pulmonary ECs (25 × upregulation). Experimental overexpression in a transgenic zebrafish line and in HUVECs was sufficient to induce spontaneous vascular leakage. To the contrary, genetic MIR155 reduction protects against permeability both in vitro and in endotoxemia in vivo in MIR155 heterozygote knockout mice thereby improving survival by 40%. A tight junction protein, Claudin-1, was down-regulated both in endotoxemia and by experimental MIR155 overexpression. Translationally, MIR155 was detectable at high levels in bronchoalveolar fluid of patients with ARDS compared to healthy human subjects.ConclusionsWe found that MIR155 is upregulated in the endothelium in mouse and men as part of a systemic inflammatory response and might contribute to the pathophysiology of vascular leakage in a Claudin-1-dependent manner. Future studies have to clarify whether MIR155 could be a potential therapeutic target.

TRAV1-2+ CD8+ T-cells including oligoconal expansions of MAIT cells are enriched in the airways in human tuberculosis

Mucosal-associated invariant T (MAIT) cells typically express a TRAV1-2+ semi-invariant TCRα that enables recognition of bacterial, mycobacterial, and fungal riboflavin metabolites presented by MR1. MAIT cells are associated with immune control of bacterial and mycobacterial infections in murine models. Here, we report that a population of pro-inflammatory TRAV1-2+ CD8+ T cells are present in the airways and lungs of healthy individuals and are enriched in bronchoalveolar fluid of patients with active pulmonary tuberculosis (TB). High-throughput T cell receptor analysis reveals oligoclonal expansions of canonical and donor-unique TRAV1-2+ MAIT-consistent TCRα sequences within this population. Some of these cells demonstrate MR1-restricted mycobacterial reactivity and phenotypes suggestive of MAIT cell identity. These findings demonstrate enrichment of TRAV1-2+ CD8+ T cells with MAIT or MAIT-like features in the airways during active TB and suggest a role for these cells in the human pulmonary immune response to Mycobacterium tuberculosis.

ЗМІНА СТАНУ КОМПЛЕКСУ ЛЕГЕНЕВОГО СУРФАКТАНТУ ПРИ ВТРУЧАННЯХ З ПРИВОДУ ГОСТРОГО АОРТАЛЬНОГО СИНДРОМУ В УМОВАХ ШТУЧНОГО КРОВООБІГУ

Objective. To study changes in the state of pulmonary surfactant complex during interventions in acute aortic syndrome. Materials and methods. Biochemical analysis and cytological study were conducted on 142 samples obtained during the bronchoalveolar lavage performed intraoperatively or in the early postoperative period. Results. It was determined that in the postoperative period the bronchoalveolar fluid of patients with disorders of gas exchange contained significantly more general protein, albumin and fibrinogen, indicating an increase in the alveolar membrane transport. The higher content of plasma proteins in the alveolar fluid of patients with ARDS was due to low phospholipid content, reflecting damage to the lung surfactant system. Conclusion. The obtained data indicates the damage of pulmonary surfactant complex followed by an increase in the alveolar membrane transport leading to development of acute respiratory distress syndrome after surgery on the aorta.

Determination of MIC Distribution and Mechanisms of Decreased Susceptibility to Bedaquiline among Clinical Isolates of Mycobacterium abscessus.

ABSTRACTChemotherapeutic options against Mycobacterium abscessus infections are very limited. Bedaquiline, a new antituberculosis (anti-TB) drug, is effective for the treatment of multidrug-resistant TB. However, few data are available on bedaquiline for treatment of M. abscessus infections. In this study, we determined the profile for in vitro susceptibility of M. abscessus clinical isolates to bedaquiline and investigated the potential molecular mechanisms of decreased susceptibility. A total of 197 M. abscessus clinical isolates were collected from sputum and bronchoalveolar fluid of patients with lung infections. Standard broth microdilution test revealed that bedaquiline exhibited high in vitro killing activity against M. abscessus isolates, with a MIC50 of 0.062 and a MIC90 of 0.125 mg/liter. Whole-genome sequencing data showed that no nonsynonymous mutation occurred in atpE, the gene encoding the bedaquiline-targeted protein. However, of 6 strains with decreased susceptibility of bedaquiline (MIC = 0.5 to 1 mg/liter), 3 strains had nonsynonymous mutations in mab_4384, the gene encoding the repressor of efflux pump MmpS5/MmpL5. Quantitative reverse transcription-PCR (qRT-PCR) analysis showed that the expression of MmpS5/MmpL5 in the group with decreased susceptibility to bedaquiline was significantly higher than in those with medium MICs (MIC = 0.125 to 0.5 mg/liter) or in the low-MIC group (MIC ≤ 0.062 mg/liter). Two isolates with increased MICs did not show overexpression of MmpS5/MmpL5, which could not be explained by known molecular mechanisms. This is the first report showing the association of MmpS5/MmpL5 with decreased bedaquiline susceptibility in M. abscessus clinical isolates and suggesting the presence of other, yet-to-be identified mechanisms for decreased bedaquiline susceptibility in M. abscessus.

Порівняння локального запалення при загостренні хозл та перитоніті з урахуванням цитокінового профілю

Introduction. Pathological signs of chronic obstructive pulmonary disease (COPD) is the destruction of the parenchyma with emphysema and inflammation of the small airways. Peritonitis – one of the most frequent pathologies in surgery. It is associated with the development of inflammation on systemic and local levels. Determining the level of proinflammatory mediators in peritoneal fluid and serum and their ratio is important for the diagnosis and prognosis of disease and choice of effective treatments. Typically, the level of inflammatory cytokines in sputum and bronchoalveolar fluid of patients with COPD is increasing. On the contrary, the content of cytokine suppressive properties may decline. In literature there is no data on the comparative characteristics of proinflammatory cytokine suppressive and locally in exacerbations of COPD and development of serous-fibrinous peritonitis.The aim of the study – to compare local levels of proinflammatory cytokine suppressive and in patients with exacerbations of COPD and serous-fibrinous peritonitis.Methods of the research. The objects of the study were samples of sputum of COPD patients and control individuals and patients with peritoneal fluid serous-fibrinous peritonitis obtained during surgery. The content of cytokines in induced sputum and peritoneal fluid was determined by ELISA using reagents production "Vector-Best".Results and Discussion. It was found that the concentration of proinflammatory cytokines such as interlekiny-1β, - 6 (IL-1β, IL-6) and tumor necrosis factor-α (TNF-α), was significantly higher than in the control group, indicating the uniformity of local inflammation during peritonitis as well as in COPD. However, in patients with peritonitis content of IL-6 and TNF-α increased relative COPD patients, indicating a higher activity of local inflammation during peritonitis. Regarding suppressive cytokines, when peritonitis concentration of IL-10 was significantly higher, and transforming growth factor-β - lower than in patients with COPD. In serum and induced sputum of patients with COPD observed increase in IL-1ta TNF-α.Conclusions. Local inflammation in COPD exacerbation and serous-fibrinous peritonitis is characterized by the simultaneous increase in proinflammatory cytokines and suppressive. In patients with peritonitis compared to COPD exacerbation, observed a higher activity of local inflammation, manifested significant increase in the level of IL-6 and TNF-α. When analyzing the cytokine profile of the definition of proinflammatory mediators and suppressive installed uniformity of local inflammation in COPD exacerbation and development of serous-fibrinous peritonitis.

Clinical Implications of Hepatocyte Growth Factor, Interleukin-20, and Interleukin-22 in Serum and Bronchoalveolar Fluid of Patients with Non-Small Cell Lung Cancer.

Hepatocyte growth factor (HGF) is involved in tumorigenesis, interleukin-20 (IL-20) is an inhibitor of angiogenesis, and interleukin-22 (IL-22) stimulates tumor growth. The aim of this study was to determine the level of HGF, IL-20, and IL-22 in both serum and bronchoalveolar lavage fluid (BALF) of non-small cell lung cancer (NSCLC) patients before onset of chemotherapy, the nature of the interrelationships between these markers, and their prognostic significance regarding post-chemotherapy survival time. We studied 46 NSCLC patients and 15 healthy subjects as a control group. We found significantly higher serum levels of HGF and IL-22 in the NSCLC patients than those in controls [pg/ml: HGF - 1911 (693-6510) vs. 1333 (838-3667), p = 0.0004; IL-22 - 10.66 (1.44-70.34) vs. 4.69 (0.35-12.29), p = 0.0007]. In contrast, concentrations of HGF and IL-22 in BALF were lower in NSCLC patients than those in controls [pg/ml: HGF - 72 (6-561) vs. 488 (14-2003), p = 0.0002; IL-22 - 2.28 (0.70-6.52) vs. 3.72 (2.76-5.64), p = 0.002]. In the NSCLC patients, there was a negative correlation between the serum level of IL-20 and time to tumor progression (r = -0.405, p = 0.04) and between the serum level of HGF and survival time (r = -0.41, p = 0.005). In addition, a higher serum level of HGF and a higher BALF level of IL-22 in patients were linked with a shorter overall survival. We conclude that HGF, IL-20, and IL-22 in the serum and BALF of NSCLC patients before chemotherapy may be a prognostic of cancer progression.

(440) - Expression of S100A4 Protein is T Cell-mediated in a Humanized Model of Obliterative Airway Disorder

Obliterative airway disorder (OAD) is a major cause of allograft dysfunction after lung transplantation (LTx). Increased levels of calcium-binding S100 proteins have been recently detected in the bronchoalveolar fluid of patients with OAD after LTx. We sought to investigate the expression of S100A4 protein in an original humanized model of OAD.

Allergen Challenge Increases Anandamide in Bronchoalveolar Fluid of Patients With Allergic Asthma

Preclinical studies suggest that the endocannabinoid anandamide restrains allergic airway obstruction and inflammation. Therefore, we applied saline or allergens via bronchoscopy into different lung segments of patients with allergic asthma. Twenty-four hours later, anandamide concentrations in bronchoalveolar lavage fluids had increased more than fourfold. Anandamide concentrations correlated with severity of airway inflammation. These observations suggest that allergen exposure specifically activates the pulmonary endocannabinoid system (ECS) in patients with allergic asthma.

OR.37. Th17 Response Polarization in Peripheral Blood and Bronchoalveolar Fluid of Patients with Chronic Obstructive Pulmonary Disease

OR.37. Th17 Response Polarization in Peripheral Blood and Bronchoalveolar Fluid of Patients with Chronic Obstructive Pulmonary Disease

In vitro synthesis of angiotensin-converting enzyme by alveolar macrophages is increased in disseminated sarcoidosis.

To determine the responsibility of alveolar macrophage (AM) for the increased concentration of angiotensin-converting enzyme (ACE) in bronchoalveolar fluid of patients with sarcoidosis, we cultured AM recovered by bronchoalveolar lavage of 30 subjects: 9 were normal control subjects (group C), 10 had intrathoracic localized sarcoidosis (group LS), and 11 had disseminated intrathoracic and extrathoracic sarcoidosis (group DS). Cells were cultured for 7 days and synthesis of ACE was evaluated according to the difference between final (ACEqe) and initial (ACEqi) ACE content. In groups C, LS, and DS, ACEqi were, respectively, 3.0 ± 1.7, 3.3 ± 1.9, and 4.8 ± 2.4 U/106 AM, and ACEqe were 6.2 ± 1.7, 6.7 ± 2.5, and 11.3 ± 2.8 U/106 AM. ACEqe was higher than ACEqi in all 3 groups (p < 0.01). When cycloheximide was added to the AM culture, ACEqe did not differ from ACEqi, in opposition to what was observed without cycloheximide (p < 0.002). ACEqi in group LS and DS did not differ from ACEqi in Group C, but ACEqe in Group DS was higher than in Groups C and LS (p < 0.01). We conclude that AM both from normal subjects and from subjects with sarcoidosis contain ACE, cultured AM synthesize ACE, and that a greater amount of ACE is produced when AM are obtained from patients with disseminated sarcoidosis.