Intracellular detection of interleukin 17 and other cytokines in human bronchoalveolar lavage fluid: a first assessment.
De Luca Anna,Rindi Laura,Celi Alessandro,Melosini Lorenza,Paggiaro Pierluigi,Nelli Luca Ceccherini,Garzelli Carlo,Freer Giulia
Immunology letters
Increasing evidence links pulmonary pathology to cytokines determining an inflammatory environment in the lung. Detection of cells secreting specific cytokines in BALF could be helpful as a diagnostic tool but which cytokines to choose among their great variety may be the first question to solve. The aim of this study was to investigate the Th1, Th2 and Th17 cytokine profile in whole cells within the human bronchoalveolar lavage fluid (BALF) by flow cytometry, with a focus on interleukin (IL)-17-producing cells, in order to assess which cytokines might lend themselves as markers of disease in future studies. BALF and paired peripheral blood samples were collected from 52 patients admitted to hospital for pulmonary pathologies. Cells obtained from BALF and peripheral blood were incubated in vitro in the absence or presence of appropriate stimuli and analyzed for intracellular content of IL-4, -10, -12, -17, interferon (IFN)γ and tumor necrosis factor (TNF)α in association to expression of either HLA-DR or CD4. IL-17-secreting cells were further characterized. Production of IL-17 by unstimulated BALF cells could be detected in 2 of the 32 patients that could be examined; upon PMA/IM stimulation in vitro, IL-17 was produced by varying percentages of lymphocytes, mostly memory CD4(+) cells, in all BALF samples. IL-4 could be detected in a relatively high proportion of unstimulated HLA-DR(+/-), SSC(hi) cells, most probably granulocytes; IL-10 could be found mostly in macrophages in a number of the BALF samples analyzed. Finally, IFNγ and TNFα were only produced by lymphocytes after in vitro stimulation. This study shows that T cells producing IL-17 can be found in the lung of respiratory patients in the absence of ex vivo stimulation, making IL-17 a good candidate marker of specific pathologies of the lung. Upon stimulation, IL-17 production was accounted for by CD4(+) CD45RO(+) cells. Other cytokines are also discussed. An interesting cytokine secretion profile found in BALF from a patient with rheumatoid lung disease is also reported.
10.1016/j.imlet.2011.10.005
IL-11 and CCL-1: Novel Protein Diagnostic Biomarkers of Lung Adenocarcinoma in Bronchoalveolar Lavage Fluid (BALF).
Pastor Maria Delores,Nogal Ana,Molina-Pinelo Sonia,Quintanal-Villalonga Álvaro,Meléndez Ricardo,Ferrer I,Romero-Romero Beatrice,De Miguel Maria José,López-Campos José Luis,Corral Jesús,García-Carboner Rocío,Carnero Amancio,Paz-Ares Luis
Journal of thoracic oncology : official publication of the International Association for the Study of Lung Cancer
INTRODUCTION:Lung cancer (LC) and chronic obstructive pulmonary disease (COPD) are smoking-related diseases, with the presence of COPD itself increasing the risk for development of LC, probably owing to underlying inflammation. LC is typically detected at late stages of the disease and carries a poor prognosis. There is an unmet need for methods to facilitate the early detection of LC in high-risk subjects such as smokers. METHODS:The expression of inflammatory proteins in bronchoalveolar lavage fluid (BALF) samples was studied by antibody arrays in a prospective cohort of 60 smokers of more than 30 pack-years divided into four groups (control, patients with LC, patients with COPD, and patients with LC plus COPD). Relevant biomarkers were validated by Western blot. Additional validation with enzyme-linked immunosorbent assay (ELISA) was carried out on two independent controlled cohorts of 139 patients (control, patients with LC, patients with COPD, and patients with LC plus COPD) and 160 patients (control and patients with LC of all histological types). RESULTS:A total of 16 differentially expressed proteins in samples from patients with LC, COPD, and LC plus COPD were identified by antibody arrays and validated by Western blot and ELISA. C-C motif chemokine ligand 1 (CCL-1) and interleukin-11 (IL)-11 were selectively expressed in samples from patients with adenocarcinoma with or without COPD (p < 0.005). These proteins exhibited a remarkable diagnostic performance for lung adenocarcinoma in an independent cohort of 139 patients. Receiver operating characteristic curves showed that the optimum diagnostic cutoff value for IL-11 was 42 pg/mL (area under the curve = 0.93 [95% confidence interval: 0.896-0.975], sensitivity 90%, specificity 86%), whereas for CCL-1 it was 39.5 pg/mL (0.83 [95% confidence interval: 0.749-0.902], sensitivity 83%, and specificity 74%). Further validation of the ELISA biomarkers at the aforementioned cutoffs was performed in an additional cohort of 160 patients (20 controls, 66 patients with LC, and 74 patients with LC plus COPD). There was a significant correlation between BALF levels of IL-11 and CCL-1 (r = 0.76, p < 0.001), and the use of both biomarkers increased the diagnostic accuracy to 96.1% in the two validation cohorts. Appropriate diagnostic performance was observed for all subgroups regardless of stage at diagnosis, involvement of the bronchial tract, pack-years smoked, and number of cells in BALF. CONCLUSIONS:IL-11 and CCL-1 are highly specific biomarkers with great accuracy for the diagnosis of lung adenocarcinoma in BALF specimens. Further study of these proteins as markers for the early diagnosis and screening of plasma and other biological materials is warranted.
10.1016/j.jtho.2016.07.026
Distribution and Expression of IL-17 and Related Cytokines in Children with Mycoplasma pneumoniae Pneumonia.
Fan Huifeng,Lu Bingtai,Yang Diyuan,Zhang Dongwei,Shi Tingting,Lu Gen
Japanese journal of infectious diseases
The pathogenesis of Mycoplasma pneumoniae pneumonia (MPP), specifically the local immune responses in the lungs, is poorly understood. In this study, flow cytometry was used to analyze IL-17 and related cytokines in plasma and bronchoalveolar lavage fluid (BALF) samples from 18 and 30 pediatric patients with general MPP (GMPP) and refractory MPP (RMPP), respectively. The levels of IL-1Ra, IL-6, IL-8, IL-17, and TNF-α were significantly elevated in the BALF of MPP children compared to the plasma (P < 0.01). Although the plasma IL-6 levels in the children with RMPP were higher than those in the children with GMPP (P < 0.05), the IL-17 levels showed the opposite trend (P < 0.05). The children with RMPP had significantly higher BALF levels of IL-8, IL-17, and TNF-α than the children with GMPP (P < 0.05), and the elevated levels of IL-17 correlated with the increased focal size of the lung lesions (P < 0.05). The elevated levels of IL-17 and related cytokines in the BALF samples could indicate that the local inflammatory response should be distinguished from the systemic inflammatory response in children with MPP. Moreover, RMPP might involve an aggravated inflammatory progression at the site of infection. The levels of IL-17 might correlate with the extent and severity of the lung lesions in MPP.
10.7883/yoken.JJID.2019.113
Evaluation of cytokines in the tumor microenvironment of lung cancer using bronchoalveolar lavage fluid analysis.
Cancer immunology, immunotherapy : CII
INTRODUCTION:Lung cancer is the leading cause of death by cancer. In recent years, immunotherapy with checkpoint inhibitors (ICI) emerged as a promising new therapeutic approach. However, a deeper understanding of the immunologic responses adjacent to the tumor known as tumor microenvironment (TME) is needed. Our study investigated TME of lung cancer by analyzing cytokines in bronchoalveolar lavage fluid (BALF). MATERIALS AND METHODS:Between January 2018 and June 2019, 119 patients were prospectively enrolled in this study. For each cancer patient, levels of 16 cytokines (fractalkine, granulocyte-macrophage colony-stimulating factor (GM-CSF), interferon gamma (IFN-γ), tumor necrosis factor alpha (TNF-α), and interleukins (IL): IL-1b, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IL-12p70, IL-13, IL-17A, and IL-23) were measured in BALF and serum and compared to healthy individuals and patients with other lung diseases. RESULTS:There were several significant differences of cytokine levels of patients with lung cancer compared to healthy individuals. However, none of them remained in the multivariate analysis compared to other lung diseases in either BALF or serum. Furthermore, there were no significant differences between the groups in cell differentiation of either BALF or serum. Cytokine levels in BALF were generally near the lower detection limit and showed almost no correlation with their respective levels measured in serum of the same individual. CONCLUSIONS:Cytokines in BALF and serum of lung cancer patients may indicate unspecific inflammation. BAL is not recommendable as a tool to investigate TME of lung cancer. Therefore, cytokines measured in BALF are probably not appropriate as predictors in patients treated with ICIs.
10.1007/s00262-020-02798-z
Exhausted and Apoptotic BALF T Cells in Proinflammatory Airway Milieu at Acute Phase of Severe Mycoplasma Pneumoniae Pneumonia in Children.
Chen Xi,Liu Fang,Zheng Baoying,Kang Xiaohui,Wang Xiaolin,Mou Wenjun,Zhang Hui,Jiao Anxia,Zhao Shunying,Gui Jingang
Frontiers in immunology
Severe mycoplasma pneumoniae pneumonia (MPP) in children presents with serious clinical complications. Without proper and prompt intervention, it could lead to deadly consequences. Dynamics of the inflammatory airway milieu and activation status of immune cells were believed to be the hallmark of the pathogenesis and progress of the disease. In this study, by employing the T-cell sorting and mRNA microarray, we were able to define the main feature of the chemokine/cytokine expression and the unique characteristics of T cells in the bronchoalveolar lavage fluid (BALF) from severe MPP patients at acute phase. Our study for the first time delineated the molecular changes in isolated BALF T cells in severe MPP children with respect to the cytokine/chemokine expression, cell activation, exhaustion, and apoptosis. By comparing the BALF aqueous expression of cytokines/chemokines with that in sorted T cells, our data give a preliminary clue capable of finishing out the possible cell source of the proinflammatory cytokines/chemokines from the BALF mixture. Meanwhile, our data provide a distinctively pellucid expression profile particularly belonging to the isolated BALF T cells demonstrating that in the inflammatory airway, overactivated T cells were exhausted and on the verge of apoptotic progress.
10.3389/fimmu.2021.760488
BALF cytokines in different phenotypes of chronic lung allograft dysfunction in lung transplant patients.
Berastegui Cristina,Gómez-Ollés Susana,Sánchez-Vidaurre Sara,Culebras Mario,Monforte Victor,López-Meseguer Manuel,Bravo Carlos,Ramon Maria-Antonia,Romero Laura,Sole Joan,Cruz Maria-Jesus,Román Antonio
Clinical transplantation
The long-term success of lung transplantation (LT) is limited by chronic lung allograft dysfunction (CLAD). Different phenotypes of CLAD have been described, such as bronchiolitis obliterans syndrome (BOS) and restrictive allograft syndrome (RAS). The purpose of this study was to investigate the levels of cytokines and chemokines in bronchoalveolar lavage fluid (BALF) as markers of these CLAD phenotypes. BALF was collected from 51 recipients who underwent (bilateral and unilateral) LT. The study population was divided into three groups: stable (ST), BOS, and RAS. Levels of interleukin (IL)-4, IL-5, IL-6, IL-10, IL-13, tumor necrosis factor alpha (TNF-α), interferon-gamma (IFN-γ), and granulocyte-macrophage colony-stimulating factor (GM-CSF) were measured using the multiplex technology. BALF neutrophilia medians were higher in BOS (38%) and RAS (30%) than in ST (8%) (P=.008; P=.012). Regarding BALF cytokines, BOS and RAS patients showed higher levels of INF-γ than ST (P=.02; P=.008). Only IL-5 presented significant differences between BOS and RAS (P=.001). BALF neutrophilia is as a marker for both CLAD phenotypes, BOS and RAS, and IL-5 seems to be a potential biomarker for the RAS phenotype.
10.1111/ctr.12898