Allergy Asthma Immunol Res.  2020 May;12(3):412-429. 10.4168/aair.2020.12.3.412.

Relationship of Microbial Profile With Airway Immune Response in Eosinophilic or Neutrophilic Inflammation of Asthmatics

• Son JH
• Kim JH
• Chang HS
• Park JS
• Park CS

• KMID: 2471226
• DOI: http://doi.org/10.4168/aair.2020.12.3.412

Abstract

PURPOSE
Different characteristics of airway microbiome in asthmatics may lead to differential immune responses, which in turn cause eosinophilic or neutrophilic airway inflammation. However, the relationships among these factors have yet to be fully elucidated.
METHODS
Microbes in induced sputum samples were subjected to sequence analysis of 16S rRNA. Airway inflammatory phenotypes were defined as neutrophils (>60%) and eosinophils (>3%), and inflammation endotypes were defined by levels of T helper (Th) 1 (interferon-γ), Th2 (interleukin [IL]-5 and IL-13), Th-17 (IL-17), and innate Th2 (IL-25, IL-33, and thymic stromal lymphopoietin) cytokines, inflammasomes (IL-1β), epithelial activation markers (granulocyte-macrophage colony-stimulating factor and IL-8), and Inflammation (IL-6 and tumor necrosis factor-α) cytokines in sputum supernatants was assessed by enzyme-linked immunosorbent assay.
RESULTS
The numbers of operational taxonomic units were significantly higher in the mixed (n = 21) and neutrophilic (n = 23) inflammation groups than in the paucigranulocytic inflammation group (n = 19; p < 0.05). At the species level, Granulicatella adiacens, Streptococcus parasanguinis, Streptococcus pneumoniae, Veillonella rogosae, Haemophilus parainfluenzae, and Neisseria perflava levels were significantly higher in the eosinophilic inflammation group (n = 20), whereas JYGU_s levels were significantly higher in the neutrophilic inflammation group compared to the other subtypes (P < 0.05). Additionally, IL-5 and IL-13 concentrations were correlated with the percentage of eosinophils (P < 0.05) and IL-13 levels were positively correlated with the read counts of Porphyromonas pasteri and V. rogosae (P < 0.05). IL-1β concentrations were correlated with the percentage of neutrophils (P < 0.05). had a tendency to be positively correlated with the read count of JYGU_s (P = 0.095), and was negatively correlated with that of S. pneumoniae (P < 0.05).
CONCLUSIONS
Difference of microbial patterns in airways may induce distinctive endotypes of asthma, which is responsible for the neutrophilic or eosinophilic inflammation in asthma.

Keyword

Asthma; eosinophils; neutrophils; microbiome; IL-13; IL-1β

MeSH Terms

Asthma
Colony-Stimulating Factors
Cytokines
Enzyme-Linked Immunosorbent Assay
Eosinophils*
Haemophilus parainfluenzae
Inflammasomes
Inflammation*
Interleukin-13
Interleukin-33
Interleukin-5
Microbiota
Necrosis
Neisseria
Neutrophils*
Phenotype
Pneumonia
Porphyromonas
Sequence Analysis
Sputum
Streptococcus
Streptococcus pneumoniae
Veillonella
Colony-Stimulating Factors
Cytokines
Inflammasomes
Interleukin-13
Interleukin-33
Interleukin-5

Figure

• 

  Fig. 1 Comparison of the OTUs numbers and the Shannon index among the four inflammatory subtypes: mixed (n = 21), Neu (n = 23), Eos (n = 20), and Pauci (n = 19). The normality of the data distribution was assessed with the Shapiro-Wilk test, between-group comparisons were performed using the Kruskal-Wallis test followed by post hoc analyses, and Mann-Whitney U tests were used to evaluate nonparametric data. All data are presented as medians with 25% and 75% quartiles.OTU, operational taxonomic unit; Eos, eosinophilic; Neu, neutrophilic; Pauci, paucigranulocytic.

• 

  Fig. 2 Clustering and heatmap for microbial species among the four inflammatory subtypes.M, mixed (n = 21); N, neutrophilic (n = 23); E, eosinophilic (n = 20); P, paucigranulocytic (n = 19).

• 

  Fig. 3 Correlations between cytokine levels and the percentages of eosinophils and neutrophils in the sputum for the overall cohort (n = 83). The IL-5, IL-13, and IL-1β values were normalized to protein levels in the sputum. The normality of the data distribution was assessed with the Shapiro-Wilk test and statistical significance was evaluated with the Spearman's rho test. Values are presented as correlation coefficients (r) with P values.IL, interleukin.

• 

  Fig. 4 Correlations between cytokine levels and bacterial normalized read counts for the overall cohort (n = 83). The normality of the data distribution was assessed using the Shapiro-Wilk test and statistical significance was evaluated with the Spearman's rho test. Values are presented as correlation coefficients (r) and with P values.IL, interleukin.

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