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Characterisation of resident alveolar macrophages in bronchoalveolar lavage fluid (BALF) in non-fumer healthy, fumer and COPD patients

ABSTRACT: Characterisation of resident alveolar macrophages in bronchoalveolar lavage fluid (BALF) in non-fumer healthy, fumer and COPD patients

PROVIDER: PRJNA762566 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR15859400.fastq.gz	Fastqsanger.gz
	SRR15859401.fastq.gz	Fastqsanger.gz
	SRR15859402.fastq.gz	Fastqsanger.gz
	SRR15859403.fastq.gz	Fastqsanger.gz
	SRR15859404.fastq.gz	Fastqsanger.gz

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Characterization of two lung alveolar macrophages in bronchoalveolar lavage fluid (BALF) cells in non-fumer healthy, fumer and COPD patients

Project description:Rationale: Alveolar macrophages (AM) are functionally important innate cells involved in lung homeostasis and immunity. However, it remains unclear whether ontogenically and functionally distinct subpopulations of AM can be found in healthy human airways, and to what extent conditions like smoking or chronic obstructive pulmonary disease (COPD) trigger changes in the AM compartment. Objective: To explore the phenotypical, ontological, transcriptional and functional diversity of human AM isolated from healthy non-smokers, healthy smokers and COPD patients. Methods: We analyzed bronchoalveolar lavage fluid (BALF) cells by flow cytometry, bulk and single-cell(sc) RNA-sequencing (RNA-seq). Main Results: We found that BALF CD206 + macrophage subpopulations could be distinguished based on their level of auto-fluorescence. CD206 + autofluorescent high (AF hi ) AM were identified as classical self-proliferative AM, while autofluorescent low (AF lo ) AM originated from monocytes and were expressing both monocyte- and classical AM-related genes. Of note, monocyte-derived AF lo AM were equally represented in each category of subjects and exhibited a functionally distinct immunoregulatory profile, including the ability to secrete the immunosuppressive cytokine interleukin-10 (IL-10). Finally, we provide evidence that the monocyte-associated marker CCR2 can be used by flow cytometry to distinguish AM according to their origin.

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Project description:Abstract: Chronic obstructive pulmonary disease (COPD) is a progressive lung disease for which there is no cure. Accumulating research results suggest a role for extracellular vesicles (EVs) in the pathogenesis of COPD. This study aimed to uncover the involvement of EVs and their molecular cargo in the progression of COPD by identification of EV-associated protein and microRNA (miRNA) profiles. We isolated EVs from bronchial alveolar lavage fluid (BALF) of 18 patients with COPD and 11 healthy controls by size exclusion chromatography. EV isolates were characterized by Nnanoparticle tracking analysis and protein content. Proteomic analysis revealed higher abundance of 284 proteins (log2FC > 1) and lower abun-dance of 3 proteins (log2FC <-1) in EVs derived from patients with COPD. Ingenuity pathway analysis showed that proteins enriched in COPD-associated EVs trigger inflammatory responses including neutrophil degranulation. Variances in surface receptors and ligands associated with COPD EVs suggested preferential interaction with alveolar cells. Small RNAseq analysis iden-tified higher abundance of ten miRNAs and lower abundance of one in EVs from COPD versus controls (Basemean >100, FDR<0.05). Our data indicate that the molecular composition of EVs in BALF of patients with COPD is altered compared to healthy control EVs. Several components in COPD EVs were identified that may perpetuate inflammation and alveolar tissue destruction.

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