Exosomes Derived from Dermatophagoides farinae Induce Allergic Airway Inflammation

Abstract

Dermatophagoides farinae , a predominant species of house dust mites in China, has displayed pathogenic role in allergic disorders, and exosomes derived from human bronchoalveolar lavage fluid have been strongly associated with allergic respiratory diseases progression. However, the pathogenic role of D. farinae -derived exosomes in allergic airway inflammation has remained unclear until now.

Dermatophagoides farinae microRNAs released to external environments via exosomes regulate inflammation-related gene expression in human bronchial epithelial cells

“…The miRNAs expression profiles in DFA-related samples. *:Expression profile of miRNAs from DFA-derived exosomes was partly published in our previous report (11).…”

“…A total of 200 dust mites were collected using this approach and stored at −80 °C for sequencing by BGI (Beijing, China). We isolated DFA-derived exosomes using the method described in our previous report (11). To obtain DFA culture supernatants, the solid culture medium of DFA was dissolved with sterile 1 × phosphate buffered saline (PBS; Gibco, Grand Island, NY, USA).…”

“…Following library construction, sRNA was sequenced with the BGISEQ-500 High-throughput Sequencing Platform (BGI; Shenzhen, China). Stringent quality control was performed at each sequencing procedure to ensure the reliability of sequencing data, and detailed sequencing procedures were described in our previous study (11). Following removal of lowquality tags, there were 46,095,191, 40,126,626 and 40,134,247 clean tags yielded from DFA mites, DFA-derived exosomes and DFA culture supernatants, respectively, and the length distribution of sRNA is displayed in Supplementary Figure S1.…”

“…The presence of these conserved and novel miRNAs was verified using qPCR assay, with templates transcribed from RNA samples of DFA mites, DFA-derived exosomes and DFA culture supernatants using the poly (A)-tailing technique (11). qPCR assay detected lower expression of dfa-miR-276-3p, dfa-miR-5735-3p, dfa-miR-252, dfa-miR-1-3p, dfa-miR-let-7b and dfa-novel-miR1, 2, 3, 4 and 5 in DFA culture supernatants than in DFA mites and DFA-derived exosomes (Figures 2A, B), with dfa-miR-276-3p as the most highly expressed conserved miRNA in DFA samples (Figure 2A), and dfa-novel-miR1, 2 and 4 presenting relatively higher expression than other novel miRNAs in DFA mites and DFA-derived exosomes (Figure 2B).…”

“…MiRNA let-7 was reported to present proinflammatory effects in experimental asthma (9), and overexpression of miR-1 reduces asthma phenotype in endothelial cells and alleviates airway eosinophilia in mouse models (10). Our previous study presented the successful isolation of DFAderived exosomes and profiled the expression of miRNAs from DFA-derived exosomes using sRNA sequencing (sRNA-Seq) (11). However, there is no knowledge on the expression profile of DFA miRNAs, and the functions of DFA miRNAs remain to be unraveled in allergy.…”