Tuberc Respir Dis.  2016 Jul;79(3):165-178. 10.4046/trd.2016.79.3.165.

Lung Microbiome Analysis in Steroid-NaÑ—ve Asthma Patients by Using Whole Sputum

• Jung JW
• Choi JC
• Shin JW
• Kim JY
• Park IW
• Choi BW
• Park HW
• Cho SH
• Kim K
• Kang HR

• KMID: 2326685
• DOI: http://doi.org/10.4046/trd.2016.79.3.165

Abstract

BACKGROUND
Although recent metagenomic approaches have characterized the distinguished microbial compositions in airways of asthmatics, these results did not reach a consensus due to the small sample size, non-standardization of specimens and medication status. We conducted a metagenomics approach by using terminal restriction fragment length polymorphism (T-RFLP) analysis of the induced whole sputum representing both the cellular and fluid phases in a relative large number of steroid naïve asthmatics.
METHODS
Induced whole sputum samples obtained from 36 healthy subjects and 89 steroid-naÑ—ve asthma patients were analyzed through T-RFLP analysis.
RESULTS
In contrast to previous reports about microbiota in the asthmatic airways, the diversity of microbial composition was not significantly different between the controls and asthma patients (p=0.937). In an analysis of similarities, the global R-value showed a statistically significant difference but a very low separation (0.148, p=0.002). The dissimilarity in the bacterial communities between groups was 28.74%, and operational taxonomic units (OTUs) contributing to this difference were as follows: OTU 789 (Lachnospiraceae), 517 (Comamonadaceae, Acetobacteraceae , and Chloroplast), 633 (Prevotella), 645 (Actinobacteria and Propionibacterium acnes), 607 (Lactobacillus buchneri, Lactobacillus otakiensis, Lactobacillus sunkii, and Rhodobacteraceae), and 661 (Acinetobacter, Pseudomonas, and Leptotrichiaceae), and they were significantly more prevalent in the sputum of asthma patients than in the sputum of the controls.
CONCLUSION
Before starting anti-asthmatic treatment, the microbiota in the whole sputum of patients with asthma showed a marginal difference from the microbiota in the whole sputum of the controls.

Keyword

Asthma; Microbiota; Polymorphism, Restriction Fragment Length; RNA, Ribosomal, 16S; Sputum

MeSH Terms

Acetobacteraceae
Asthma*
Consensus
Healthy Volunteers
Humans
Lactobacillus
Lung*
Metagenomics
Microbiota*
Polymorphism, Restriction Fragment Length
Propionibacterium
Pseudomonas
RNA, Ribosomal, 16S
Sample Size
Sputum*
RNA, Ribosomal, 16S

Figure

• 

  Figure 1 Diversity of microbiota profile in the healthy control and asthma groups. Diversity of microbiota did not show a distinction between healthy controls and asthma patients (p=0.937).

• 

  Figure 2 Analysis of similarity test for global community composition (ANOSIM test). Microbial composition showed a significant difference between healthy controls and asthma patients (Global R value=0.148, p=0.002).

• 

  Figure 3 Cluster analysis. Hierarchical cluster analysis indicated no distinct difference in the operational taxonomic units in the sputum samples between the two groups.

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  Figure 4 Nonmetric multidimensional scaling (nMDS). nMDS indicated no distinct difference in the operational taxonomic units in the sputum samples between the two groups.

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  Figure 5 Operational taxonomic units (OTUs) with at least 1.5-fold differences in sputum microbiota between the controls and asthma patients and fold abundance.

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