Abstract

BackgroundTuberculosis (TB) remains a debilitating, deadly disease that warrants innovative research tools to fully understand the pathogenesis and host immune responses, particularly at the site of infection and disease. In this regard, bronchoscopies with bronchoalveolar lavage (BAL) serve as a valuable technique for site of disease sample retrieval for further clinical- and basic research. Here we investigate the feasibility of research bronchoscopies in a low/middle-income area, where TB remains rife, and assess the value of retrieved BAL cells (BALC) for downstream fluorescent-based cellular evaluations.MethodsUsing quantitative and qualitative methods, we evaluate the outcomes, safety, tolerability, participant -perception and -experience, while also providing insight into participant recruitment and screening processes of our study. Using light microscopy differential counting for BALC analysis, we evaluate the cellular composition of BAL fluid (BALF) from TB patients, healthy community controls and patients with other lung diseases. We also use flow cytometry to describe the challenges associated with fluorescence-based phenotypic analysis of autofluorescent BALC.ResultsOur findings suggest that research bronchoscopies are safe, acceptable procedures for research participants and are indeed a feasible technique for future study design. We also suggest that the majority of participants are receptive to the proposition of a second research bronchoscopy. This poses an important avenue for research entailing follow-up investigations of the same study participant. Furthermore, our results show that smoking is characterized by retrieval of BALC containing particulate matter, that interferes with fluorescence-based flow cytometry data analysis. Based on light microscopy differential cell counting, our findings suggest that there are differences in the cell yields and cellular composition of the BALF between TB patients, healthy community controls and patients with other lung diseases. We also report on subject characteristics and demographic factors, namely gender and age, that have the potential to affect cell yields and cellular data of BALF.ConclusionsThese findings will serve as a valuable reference for appropriate planning and design of studies involving clinical bronchoscopies for TB and lung disease research.

Highlights

  • Bronchoscopies are widely performed as part of the clinical workup of lung disease, including the diagnosis and grading of cancerous lung lesions and identification of lung pathogens

  • We summarize our assessment of the feasibility of research bronchoscopies and provide expected reference ranges for bronchoalveolar lavage (BAL) cell (BALC) and BAL fluid (BALF) retrieval and composition from TB patients, healthy community controls and patients suffering from non-TB related lung disease in the context of the social, economic, and health setting of the Western Cape region, a setting with both affluent and poor socio-economic societies

  • We report significantly increased BAL cells (BALC) yields, decreased macrophage/monocyte frequencies, increased neutrophil frequencies and decreased lymphocyte frequencies in TB patients, relative to healthy and Other lung disease (OLD) control cohorts

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Summary

Introduction

Bronchoscopies are widely performed as part of the clinical workup of lung disease, including the diagnosis and grading of cancerous lung lesions and identification of lung pathogens. It is thought that the cellular patterns in bronchoalveolar lavage (BAL) fluid (BALF) reflect immune processes and host responses within the lung airways, its increasing popularity in the pulmonary research field [1,2,3,4,5]. Tuberculosis (TB) remains a debilitating, deadly disease that warrants innovative research tools to fully understand the pathogenesis and host immune responses, at the site of infection and disease In this regard, bronchoscopies with bronchoalveolar lavage (BAL) serve as a valuable technique for site of disease sample retrieval for further clinical- and basic research. We investigate the feasibility of research bronchoscopies in a low/middle-income area, where TB remains rife, and assess the value of retrieved BAL cells (BALC) for downstream fluorescent-based cellular evaluations

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