Search Results (263)

Human herpesvirus 6 (HHV-6) is a common virus that can reactivate in immunocompromised patients, including lung transplant (LT) recipients. This study aimed to evaluate the clinical and functional implications of HHV-6 infection in LT patients through a retrospective analysis of 175 individuals who underwent lung transplantation at the City of Health and Sciences of Turin between 2014 and 2023. Surveillance bronchoscopies—including bronchoalveolar lavage (BAL) and transbronchial biopsies—were performed at scheduled intervals over a two-year period to detect HHV-6 and other pathogens, and to assess acute rejection. Spirometries were performed to evaluate graft function. Among the cohort, 33% of 822 BAL samples tested were positive for HHV-6, with a notable association between high viral load (≥500 copies/mL) and the development of post-transplant lymphoproliferative disorder (PTLD) (13% vs. 1%, p = 0.02) at 1 month and (9% vs. 1%, p = 0.026) at 12 months. Co-infection with CMV (78% in positives vs. 55% in negatives; p = 0.006), Epstein–Barr virus (EBV) (35% vs. 16%; p = 0.010), and bacterial and fungal infection (specifically, a higher rate of isolation of Achromobacter xylosoxidans (13%), p = 0.010) was frequently observed in conjunction with HHV-6 positivity. Notably, patients with at least one HHV-6 positive BAL exhibited a significant reduction in forced vital capacity (FVC) at multiple follow-up points, FVC 82% in positives vs. 92% in negatives (p = 0.038) at 4 months and 87% vs. 98% p = 0.033 at 8 months and 87% vs. 99% p = 0.038 at 24 months. No direct associations with acute rejection or overall survival were found. By means of this study, we provide a wide overview of HHV-6 in lung transplant recipients, filling in a gap of evidence in the field. We report a remarkable incidence and a significant association with acknowledged clinically relevant viral infections, PTLD, and functional tests decline, with no association with mortality. Full article

Background/Objectives: Obesity and hyperglycemia predispose patients to respiratory infections. Although the lung is a major organ to utilize glucose, pulmonary glucose homeostasis in type 2 diabetic (T2Dx) subjects remains poorly characterized. We hypothesized that pulmonary glucose transport would be altered during T2Dx, which would be rescued with long-term metformin treatment. Methods: T2Dx was induced by feeding mice a high-fat diet for 16 weeks, with metformin treatment administered during the final 8 weeks. Results: Glucose transporter (GLUT) protein expression and trafficking was quantified by Western blotting and the biotinylated photolabeling assay, respectively. T2Dx mice exhibited obesity, and increased glucose levels in blood and bronchoalveolar lavage (BAL) fluid. T2Dx also significantly decreased protein expression of GLUTs from Class I (i.e., GLUT-2 and -4) and class III (i.e., GLUT-10 and -12) isoforms in lung. Metformin treatment restored the protein expression of GLUT-2, -4, and -10, but not GLUT-12. Pulmonary cell surface expression of GLUT-4 and -8 was also significantly reduced in T2Dx mice and rescued by metformin. Conclusions: These findings suggest that alterations in pulmonary GLUT expression and trafficking during diabetes could contribute to the elevated airway glucose levels and severity of respiratory infections. Metformin treatment restored pulmonary glucose transport during T2Dx. Full article

Background: Protracted bacterial bronchitis (PBB) and bronchiectasis share common clinical and microbiological features, but direct comparative data in children are limited. Objectives: To compare bronchoalveolar lavage (BAL) microbiology between pediatric PBB and bronchiectasis and identify predictors of lower airway and polymicrobial infections. Methods: We retrospectively reviewed children diagnosed with PBB or bronchiectasis at a tertiary center (January 2019–June 2025) who underwent both high-resolution computed tomography of the chest and bronchoscopy with BAL within a 6-month period. Multivariable logistic regression was used to identify predictors of lower airway and polymicrobial infections, adjusting for age, gender, tracheomalacia/bronchomalacia, asthma, and Bhalla score. Results: Among 135 children (85 with PBB, 50 with bronchiectasis), those with bronchiectasis were older (median 7.8 vs. 4.2 years, p < 0.001), while comorbidities showed statistically non-significant differences. The prevalence of lower airway infection was high (PBB 81.2%, bronchiectasis 74.0%; p = 0.330). Pathogen distribution demonstrated statistically non-significant differences between groups after adjustment, with Haemophilus influenzae being the most common pathogen in both groups. Moraxella catarrhalis was more frequent in PBB in unadjusted analysis (21.2% vs. 8.0%; p = 0.045), but this difference did not persist after adjustment. Polymicrobial infection occurred in 25.9% of PBB and 16.0% of bronchiectasis cases (p = 0.180). In regression analyses, younger age independently predicted polymicrobial infection (adjusted OR 0.81, 95% CI 0.69–0.95). Conclusions: BAL microbiology showed statistically non-significant differences between PBB and bronchiectasis, supporting the concept of a disease continuum. Younger children were more prone to polymicrobial infection. These findings support early targeted antibiotic therapy and close clinical surveillance to prevent progression to irreversible airway damage. Full article

Background: Hypersensitivity pneumonitis (HP) is an interstitial lung disease (ILD) characterized by inflammation of the lung parenchyma, alveoli and bronchioles induced by inhalation of organic compounds. Bird-related-HP (BRHP) is the most common type of HP, occurring in susceptible people in regular contact with birds, although a genetic susceptibility is unclear. This study investigates the impact of environmental volatile organic compounds (VOCs) on the development of HP and other pulmonary diseases, and their relationship with pulmonary inflammatory cell composition and patient outcomes. Methods: Geospatial environmental levels of VOCs (benzene, toluene, ethylbenzene, m,p-xylene and o-xylene) in patients’ homes were related to bronchoalveolar lavage (BAL) leukocyte profiles analyzed by flow cytometry of 1515 patients with different lung diseases in the region of Murcia (southeastern Spain). Results: Ethylbenzene levels over the threshold limit of 10 µg/m³ (EB10) were associated with HP (23.9% vs. 15.2%, p < 0.05). A strong association with HP was observed in patients in contact with birds living in areas with EB10 (63.0% vs. 27.4%, p < 0.001). Linear regression analysis showed that age (B = −0.058, p < 0.012), smoking (B = −0.125, p < 0.001), bird contact (B = 0.275, p < 0.001) and EB10 (B = 0.109, p < 0.001) were independent variables associated with HP. In HP patients, BAL CD4/CD8-ratio > 1.5 was associated with shorter overall survival (8.9 years vs. not-reached, p < 0.011), probably due to lower CD8+ T-lymphocyte counts observed in HP fibrotic patients (11.65 ± 2.8% vs. 23.6 ± 2.9%, p = 0.008) and in those who died during follow-up (10.0 ± 1.9% vs. 23.8 ± 2.7%, p = 0.012), suggesting a protective role for CD8+ T cells. Conclusions: High environmental ethylbenzene is strongly associated with BRHP. CD8+ T-lymphocytes could have a protective role in HP, preventing fibrosis and increasing overall survival. Full article

Aspergillosis includes a variety of diseases caused by species of the genus Aspergillus, ranging from non-invasive allergic diseases to chronic, invasive pulmonary infections, which are potentially fatal in immunocompromised hosts. Therefore, there is an urgent need for new diagnostic tools and the optimization of existing tests to improve patient care. This work reviews the most commonly used molecular methods for the diagnosis of aspergillosis from clinical samples, emphasizing their advantages. These methods included HTS, NTS, ISH, microarrays, PCR-RFLP, LAMP, and PCR in various modalities (qPCR, multiplex PCR, nested PCR, RT-PCR, endpoint PCR, U-dHRM, and ddPCR). The review showed that the most commonly used methods for diagnosing aspergillosis are NGS and PCR in their different modalities; however, each method has advantages and disadvantages. qPCR is the method that has demonstrated the greatest sensitivity and specificity on clinical samples (such as blood, serum, bronchoalveolar lavage [BAL], tissue, or sputum), since it detects specific sequences, and the validation of this method shows greater progress in achieving this objective. Likewise, NGS showed that BAL is the most suitable sample, with a higher fungal load than sputum or blood. On the other hand, NGS is not a targeted technique, since it sequences all the genetic material present. Additionally, the sensitivity for detecting pathogens decreases when clinical samples are used due to the high background of nucleic acids present in the human host. Full article

Background: Gastroesophageal reflux disease (GERD) has been implicated in recurrent wheezing, but mechanisms and diagnostic markers remain debated. Multichannel intraluminal impedance-pH (MII-pH) monitoring improves reflux detection compared to pH-metry, while bronchoalveolar lavage (BAL) cytology may provide evidence of aspiration-related airway inflammation. Objectives: This study aims to examine the relationship between reflux characteristics, BAL cytology and clinical outcomes in preschool children with severe recurrent wheeze. Methods: Preschool-aged children undergoing combined MII-pH and bronchoscopy for severe recurrent wheeze were included. BAL samples were assessed for lipid-laden macrophages (LLM). Associations between reflux parameters, BAL cytology and response to antireflux treatment were analysed. Results: GERD was identified in 70% of participants, with weakly acidic and proximal reflux episodes predominating. Children with GERD exhibited significantly higher percentages of LLM compared with those without GERD (12% vs. 1%, p < 0.001). LLM percentage correlated with multiple reflux characteristics, including weakly acidic, liquid and proximal reflux (p < 0.047; p < 0.047 and p < 0.047, respectively), as well as symptom indices (p < 0.001). Following antireflux therapy, wheezing episodes were substantially reduced. Conclusions: GERD, particularly weakly acidic and proximal reflux, is associated with airway inflammation and recurrent wheeze in preschool children. BAL LLM percentage may serve as a surrogate marker of reflux-related microaspiration. MII-pH monitoring enhances diagnostic accuracy beyond pH-metry and may help guide targeted antireflux interventions. Full article

Background/Objectives:  Pneumocystis jirovecii pneumonia (PJP) remains a major cause of morbidity and mortality in immunocompromised patients. Bronchoalveolar lavage (BAL) is the diagnostic gold standard but is invasive and often impractical in critically ill patients. Oropharyngeal wash (OW) polymerase chain reaction (PCR) offers a rapid, non-invasive alternative. We performed a systematic review focusing on this respiratory sample’s diagnostic accuracy and clinical utility. Methods: We searched PubMed, Scopus, Web of Science, Cochrane Library, and clinical trial registries including ClinicalTrials.gov and MedRxiv for studies of PCR-based P. jirovecii detection in OW samples from immunocompromised adults, using BAL or induced sputum as reference standards. The Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) methodology was followed. Quality was assessed with Quality Assessment of Diagnostic Accuracy Studies-2 (QUADAS-2), and pooled sensitivity/specificity were estimated using a bivariate random-effects model. Results: Twelve studies (n = 633; 346 confirmed PJP cases) met the inclusion criteria. Most cohorts were human immunodeficiency virus (HIV)-positive. Pooled sensitivity was 68.3% (95% CI: 59.2–75.9) and specificity 91.8% (95% CI: 85.9–95.3); the area under the summary receiver operating characteristic curve (AUC) was 0.887. Diagnostic yield improved with pre-sample cough induction, 60-s gargling, early sampling before extended therapy, and higher fungal loads. Risk of bias was low, and no significant publication bias was detected. Conclusions: OW-based PCR delivers high specificity and moderate sensitivity for PJP diagnosis, offering a safe, scalable, and patient-friendly alternative when invasive testing is unfeasible. Optimizing collection protocols and expanding evaluation to non-HIV immunosuppressed populations could enhance its role as an early screening tool, enabling faster treatment decisions and reducing unnecessary antimicrobial exposure. Full article

Acute rejection and infections are the most frequent complications in the first year after lung transplantation, often representing relevant causes of death. There is still no consensus on the ideal strategy for preventing these events, with a still open debate on active bronchoscopic surveillance protocols vs. clinically mandated ones. The aim of our single-center exploratory study was to evaluate retrospectively the role of microbiology at bronchoalveolar lavage (BAL) at the first month from transplantation in asymptomatic patients in relation to the development of complications up to 12 months from surgery. We collected data from 28 patients who underwent surveillance bronchoscopies according to our center protocol (transbronchial biopsies and BAL at months 1, 4, 8, 12, 18, and 24 post-transplantation) who had a 12-month follow-up. The inclusion criterion was the absence of infiltrates at 1-month post-transplantation chest CT. We excluded patients transplanted due to suppurative diseases of the lung to minimize the pre-transplantation risk factors for infection. We also assessed differences in complications according to the underlying disease. We enrolled 15 patients with interstitial lung diseases (ILDs) and 13 with chronic obstructive pulmonary disease (COPD). Of the 28 patients, 11 had a positive BAL for bacteria. Patients with a positive BAL developed a higher number of pulmonary infectious complications (odds ratio of 18.33, p-value = 0.013 at regression model), with a near significance for moderate–severe pulmonary infections (odds ratio 4.8, p-value = 0.061). We did not find a significant correlation with rejection, cytomegalovirus reactivation, or pseudomembranes. We did not find differences in the rates of complications when grouping subjects according to pre-transplantation disease. Our results suggest a possible role for BAL positivity for bacteria in asymptomatic patients at surveillance bronchoscopy in predicting the development of future infections, warranting a tailored follow-up of patients that considers this data. Larger, multicentric studies are needed to explore and confirm the utility of our findings. Full article

Introduction: With advances in surgical techniques and immunosuppressive therapies, lung retransplantation has become a viable option for patients experiencing graft failure. However, retransplantation is associated with inferior clinical outcomes, and infection remains a leading cause of morbidity and mortality in lung retransplant recipients. Objectives: This study examined clinical characteristics, outcomes, and microbial spectra of respiratory infections in lung retransplant recipients. Methods: This retrospective case–control study, conducted at two tertiary care centers, included 10 lung retransplant patients and 20 matched primary lung transplant patients. Respiratory pathogens identified using bronchoalveolar lavage (BAL) were compared between two groups over a two-year post-transplantation period. Results: Pulmonary hypertension was more prevalent in the retransplant group (p = 0.030). Five-year mortality and infection-related mortality were higher in the retransplant group (both p = 0.015), along with longer hospital and intensive care unit stays (p = 0.035 and 0.017, respectively). BAL cultures revealed distinct temporal patterns: Elizabethkingia predominated early (31.6% within 1 month) in the retransplant group, with Pseudomonas increasing gradually. The primary transplant group demonstrated a more heterogeneous distribution, with Acinetobacter, Pseudomonas, and Enterococcus detected early. Conclusions: Retransplant recipients exhibited worse clinical outcomes and a distinct temporal distribution of respiratory pathogens. Particularly, the high incidence of Elizabethkingia in lung retransplant recipients highlights the need for center-specific infection surveillance and tailored preventive strategies to improve retransplantation outcomes. Full article

The use of immune checkpoint inhibitors (ICIs) has increased exponentially in recent years, leading to a significant impact on cancer patient survival. However, their administration can trigger immune-mediated adverse effects, notably pulmonary toxicity, which is a potentially serious complication. ICI-induced pneumonitis has a variable incidence ranging from 5 to 19% and usually appears in the first few months of treatment. The diagnosis requires a high index of suspicion, especially in patients with risk factors (elderly male smokers with squamous cell lung cancer, previous respiratory or autoimmune disease, and receiving combination treatment with other ICIs or chemo-radiotherapy). Chest computed tomography (CT) is a key test, allowing the identification of different radiological patterns. This study can be completed with bronchoscopy with bronchoalveolar lavage (BAL) to rule out infection or tumour progression. In general terms, treatment is based on discontinuing the causative drug, with or without the initiation of systemic corticosteroids, escalating to immunosuppressants depending on the severity and/or refractoriness of the condition. This paper provides an updated narrative review of ICI pulmonary toxicity, addressing its pathophysiology, different types of lung damage, diagnostic and therapeutic algorithms, and the emerging role of biomarkers such as KL-6 or IL-6. This article emphasises the need for a multidisciplinary approach and further prospective studies to optimise the management and prognosis of this immune-mediated complication. Full article

Verruconis species are thermophilic, darkly pigmented fungi commonly found in hot environments. Despite their environmental ubiquity, fewer than fifty human infections have been reported, with V. gallopava responsible for most cases. While infections primarily occur in immunocompromised individuals, only six cases in immunocompetent patients have been documented. We describe a case of pulmonary Verruconis infection in a 75-year-old immunocompetent woman. Despite broad-spectrum antifungal treatments, including liposomal amphotericin B and voriconazole, the patient’s condition deteriorated. Bronchoalveolar lavage (BAL) revealed hyphal forms, and fungal culture identified a Verruconis species. Antifungal susceptibility tests showed low minimal inhibitory concentrations (MICs) for amphotericin B (1 μg/mL) and voriconazole (0.5 μg/mL). Clinical manifestations of Verruconis infection in immunocompetent pneumonia patients are non-specific. Structural lung disease was identified as the primary risk factor in such hosts. BAL fungal cultures and metagenomics are valuable tools in diagnosing rare fungal infections. Treatment regimens vary, with amphotericin B and triazoles being the most commonly used antifungal agents. Currently, there are no standardized guidelines for diagnosis or treatment. Further studies are needed to establish clinical protocols. Full article

Background: Scedosporium apiospermum is a filamentous fungus increasingly recognized as an opportunistic pathogen in immunocompromised hosts, though rare infections in immunocompetent individuals with structural lung disease have been reported. Its diagnosis and management remain challenging due to non-specific clinical presentation and intrinsic resistance to multiple antifungal agents. Case Presentation: We report the case of a 66-year-old immunocompetent woman with idiopathic bilateral non-cystic fibrosis bronchiectasis, who presented with subacute cough and increased sputum production. Chest high-resolution CT revealed new subsolid and ground-glass infiltrates superimposed on stable bronchiectatic changes. Bronchoalveolar lavage (BAL) cultures isolated S. apiospermum as the sole pathogen. The patient was treated with oral voriconazole (200 mg BID) for 4 weeks, followed by a 4-week course of aerosolized amphotericin B. Clinical and radiological improvement was observed, and no relapse occurred during follow-up. Discussion: This case highlights the potential for S. apiospermum to cause clinically relevant pulmonary infection in structurally abnormal but immunocompetent lungs. Non-CF bronchiectasis may facilitate fungal colonization due to impaired mucociliary clearance and chronic mucus retention. Combined antifungal therapy involving systemic voriconazole and inhaled amphotericin B (though not yet standardized) was employed based on clinical rationale and the available literature, resulting in favorable outcomes. Conclusions:S. apiospermum pulmonary infection, although rare in immunocompetent hosts with bronchiectasis, should be considered in cases of new or persistent infiltrates. Early recognition and individualized antifungal strategies, including the potential role of inhaled agents, may improve clinical outcomes. This case reinforces the importance of multidisciplinary collaboration in the management of complex fungal infections in chronic airway disease. Full article

Mild–moderate equine asthma (MEA) is a very common but underdiagnosed pulmonary disease in horses, with mild cases not showing clinical respiratory signs. This study evaluates the influence of a standardized lunging exercise test (SLET) on bronchoalveolar lavage fluid (BALF) cytology in MEA horses. We hypothesized that SLET would increase the total nucleated cell count (TNCC) and/or percentages of inflammatory cells associated with EA. In a prospective, randomized, non-blinded, between-subjects study design of two independent groups, 39 horses (17 mild and 22 moderate) were included. They were chosen out of a cohort of horses undergoing respiratory investigations (history, clinical examination, and clinical pathology (white blood cells (WBC) and arterial blood gas analysis (aBGA)) consistent with MEA, using a scoring system in a clinical setting of an equine referral clinic. Bronchoalveolar lavage (BAL) was performed 30 min post-SLET in 16 randomly chosen horses. The other horses underwent BAL without SLET. The SLET resulted in a statistically significant increase (p < 0.001) in the proportions of neutrophils in BALF cytology, and in an increased chance of confirmation of the presumed diagnosis in horses with mild phenotypes (p < 0.001, OR = 8.00, CI = 1.28–50.04), while moderate phenotypes were less frequently diagnosed. Exercise had no association with cytology across all horses. Unexpectedly, the SLET group of horses with a moderate phenotype had a statistically significant lower TNCC (p = 0.035). In conclusion, an SLET prior to BAL might increase the chance of an MEA diagnosis. Full article

Chronic obstructive pulmonary disease (COPD) and asthma are characterised by increased oxidative stress in the lungs. The precise contribution of this stress to COPD aetiology remains unclear, partly due to the confounding influence of physiological ageing. Previous reports of increased oxidative stress in bronchoalveolar lavage (BAL) samples from individuals with COPD may at least in part be attributable to the subjects’ age. This study investigated whether increased metal concentrations at the air–lung interface would contribute to oxidative stress in the lungs. We analysed BAL samples from young and old never-smokers, young asthmatic never-smokers, older smokers without COPD and COPD patients (both current and ex-smokers). Inductively coupled plasma mass spectrometry (ICP-MS) was used to quantify a range of transition metals, including iron, copper, zinc, arsenic and cadmium. BAL concentrations of copper and zinc were significantly lower in young groups compared to the older groups, irrespective of smoking status or disease (p < 0.001 for both). BAL copper was significantly associated with several markers of oxidative stress, all of which were elevated with age: glutathione disulphide (ρ = 0.50, p < 0.001), dehydroascorbate (ρ = 0.67, p < 0.001) and 4-Hydroxynonenal (ρ = 0.43, p < 0.001). These data indicate that age-related increases in respiratory tract copper concentrations contribute to elevated levels of oxidative stress at the air–lung interface independently of respiratory disease. Full article

Background/Objectives: The quality of clinical studies is largely determined by the bioanalytical methods used for testing study samples. Rigorous assay validation following defined criteria, for example, the European Medicines Agency guideline for bioanalytical method validation, is a prerequisite for such assays. Alpha1-antitrypsin (AAT) measurement, i.e., the specific measurement of AAT protein and its associated elastase-inhibitory activity, is an integral part of assay panels for clinical studies addressing AAT deficiency. Specifically, AAT must be measured in the matrix of citrated human plasma as well as in diluted solutions with high salt concentrations obtained through bronchoalveolar lavage (BAL). Sensitive and selective measurement methods are required, as BAL has a low level of AAT. Methods: We present the validation data obtained for three AAT measurement methods. Two of them, nephelometry and the enzyme-linked immunosorbent assay, which clearly differ in their sensitivity, provide AAT protein concentrations. The third is the highly sensitive, newly developed elastase complex formation immunosorbent assay that specifically measures the inhibitory activity of AAT against its pivotal target, protease neutrophil elastase. Using samples with relevant AAT concentrations, we addressed the assays’ characteristics: accuracy, precision, linearity, selectivity, specificity, limit of quantification and short-term analyte stability Results: Overall, the three methods demonstrated low total errors, a combined measure reflecting accuracy and precision, even at low analyte concentrations of less than 0.5 µg/mL; adequate linearity over the required assay range; and acceptable selectivity and specificity. Furthermore, the short-time stability of the analyte was also demonstrated. Conclusions: All three AAT measurement methods met the acceptance criteria defined by the guidelines on bioanalytical assay validation, qualifying these methods for clinical sample analysis. Full article