Asia Pac Allergy.  2014 Oct;4(4):212-221. 10.5415/apallergy.2014.4.4.212.

Innate lymphoid cells and cytokines of the novel subtypes of helper T cells in asthma

• Sherkat R
• Yazdani R
• Ganjalikhani Hakemi M
• Homayouni
• Farahani R
• Hosseini M
• Rezaei A

• KMID: 2397102
• DOI: http://doi.org/10.5415/apallergy.2014.4.4.212

Abstract

BACKGROUND
In this study, the expression of interleukin-9 (IL-9), IL-17, IL-22, and IL-25 genes that might be the potential predisposing factors for asthma as well as count of innate lymphoid cells (ILCs) as another source of inflammatory cytokines have been evaluated.
OBJECTIVE
The aim of this study was to evaluate the expression of newly identified helper T cells signature cytokines and amount of ILCs.
METHODS
Blood and sputum samples from 23 patients with moderate to severe asthma and 23 healthy volunteers were collected. The types of allergens to which our patients were sensitive were defined using immunoblotting method. Gene expression of studied cytokines was evaluated using quantitative transcription-polymerase chain reaction and ILCs were counted by the flow cytometry method.
RESULTS
In this research, the gene expressions of IL-9, IL-17, IL-22, and IL-25 were significantly higher in asthmatics, especially in the severe form of the disease. This increase was even higher in serum samples compared with sputum samples. Counting ILCs revealed their increase in comparison with normal people.
CONCLUSION
We showed the importance of IL-25, IL-22, IL-17, and IL-9 cytokines in patients with asthma as their expression levels are increased and these increase are correlated with the severity of the disease. We also showed that the increased amount of ILCs in asthmatics could confirm their potential role in the immunopathogenesis of asthma as another source of inflammatory cytokines.

Keyword

Asthma; Innate lymphoid cell; Inteleukin-9; Inteleukin-17; Inteleukin-22; Inteleukin-25

MeSH Terms

Allergens
Asthma*
Causality
Cytokines*
Flow Cytometry
Gene Expression
Healthy Volunteers
Humans
Immunoblotting
Interleukin-17
Interleukin-9
Lymphocytes*
Methods
Sputum
T-Lymphocytes, Helper-Inducer*
Allergens
Cytokines
Interleukin-17
Interleukin-9

Figure

• 

  Fig. 1 Average expression of interleukin (IL) 9, IL-17, IL-22, and IL-25 cytokines in asthma patients and normal controls, using quantitative transcription-polymerase chain reaction. (A) Average expression of IL-9, IL-17, IL-22, and IL-25 cytokines in serum samples of cases was significantly increased compared with controls (p < 0.05). (B) Average expression of IL-9, IL-17, IL-22, and IL-25 cytokines in sputum samples of cases was significantly higher than controls (p < 0.05). (C) Comparison of the expression of IL-9, IL-17, IL-22 and IL-25 cytokines in serum and sputum samples of the asthmatics showed that the increase is much greater in serum samples (p < 0.05). (D) Comparison of the expression of IL-9, IL-17, and IL-25 cytokines in serum and sputum samples of the normal controls revealed no significant differences. However it was significant for IL-22 expression level. *p < 0.05, statistical significance.

• 

  Fig. 2 Comparing the average cytokines expression in serum and sputum samples of patients with severe and moderate asthma. Transcript levels of IL-9, IL-17, IL-22, and IL-25 cytokines in serum samples of severe asthma is significantly higher in comparison with moderate asthma (p < 0.05). In sputum samples, only IL-17 expression is not significantly more in severe asthma. *p < 0.05, statistical significance.

• 

  Fig. 3 Flow cytometry analysis of the Lin-CD127+CD161+ innate lymphoid cells (ILCs) in the peripheral blood of the asthma patients compared with the healthy individuals. (A) Forward scatter/side scatter cannels diagram of peripheral blood cells shows gated area for lymphoid cells. (B) Flow cytometry analysis of the peripheral blood samples in which T cells, B cells, and natural killer cells are excluded as one population using the fluorescein isothiocyanate-conjugated anti-CD3, anti-CD19, and anti-CD56 antibodies. Lin-/CD127+ are seen in upper left area. (C) The diagram shows the Lin-CD127+CD161+ ILCs count in an asthma patient, which appears in upper right area. (D) Diagram of the Lin-CD127+CD161+ ILCs count in a normal control. These cells are in upper right area.

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