Allergy Asthma Immunol Res.  2016 May;8(3):216-222. 10.4168/aair.2016.8.3.216.

Allergen-Dependent Differences in ILC2s Frequencies in Patients With Allergic Rhinitis

• Fan D
• Wang X
• Wang M
• Wang Y
• Zhang L
• Li Y
• Fan E
• Cao F
• Van Crombruggen K
• Zhang L

• KMID: 2391041
• DOI: http://doi.org/10.4168/aair.2016.8.3.216

Abstract

PURPOSE
Group 2 innate lymphoid cells (ILC2s) are a novel population of lineage-negative cells that induce innate type 2 responses by producing the critical Th2-type cytokines IL-5 and IL-13 in response to IL-25 and IL-33 stimulation. ILC2s accumulation in the peripheral blood of patients with allergic rhinitis (AR) is controversial; the precise role of ILC2s in the immunopathogenesis of AR is still not clear. We investigated the role of ILC2s in phenotypic AR sensitized to distinct allergens.
METHODS
Flow cytometric analysis of the peripheral blood of 7 healthy controls (HCs), 9 patients monosensitized to house dust mite (HDM), and 8 patients monosensitized to mugwort was performed to quantify ILC2s frequency. Peripheral blood mononuclear cells (PBMCs) were isolated from HDM-AR and mugwort-AR patients, and Lineage- and Lineage+ cells were separated using a fluorescence-activated cell sorter (FACS). IL-5 and IL-13 levels in the supernatants of PBMCs, and Lineage- and Lineage+ cells stimulated with IL-25 and/or IL-33 combined with IL-2 in vitro were assessed using the Milliplex magnetic bead kit.
RESULTS
The percentage of ILC2s was significantly elevated in HDM-AR patients compared to mugwort-AR patients and HCs, while no significant difference was found between mugwort-AR patients and HCs. IL-33+/-IL-25 plus IL-2 induced a significantly greater release of IL-5 and IL-13 in the PBMCs of HDM-AR patients compared to PBMCs of mugwort-AR patients. IL-25 plus IL-2 also induced a significantly greater release of IL-13 in the PBMCs of HDM-AR patients compared to PBMCs of mugwort-AR patients. Stimulation with IL-33 and/or IL-25 combined with IL-2 also induced a significantly greater IL-5 and IL-13 release from Lineage- cells compared to Lineage+ cells.
CONCLUSIONS
AR patients sensitized to HDM or mugwort allergen have distinct phenotypic and functional profiles in ILC2s frequencies. ILC2s mediate major type 2 immunity in the development of HDM-AR and may be a potential therapeutic target.

Keyword

ILC2; IL-33; IL-25; group 2 innate lymphoid cell; allergic rhinitis

MeSH Terms

Allergens
Artemisia
Cytokines
Humans
Interleukin-13
Interleukin-2
Interleukin-5
Lymphocytes
Pyroglyphidae
Rhinitis*
Allergens
Cytokines
Interleukin-13
Interleukin-2
Interleukin-5

Figure

• 

  Fig. 1 Sequential gating strategy for peripheral blood ILC2s. Lymphocytes were detected from peripheral blood mononuclear cells (left) and lineage-negative cells gated (middle). Lineage-negative lymphocytes were further assessed for the expression of CD127 and CD294 (CRTH2) or isotype control staining (right). ILC2s were identified as lineage-negative CD127+CRTH2+ lymphocytes (The example shown is typical for a patient with HDM-AR).

• 

  Fig. 2 Percentage of Lineage-CD127+CRTH2+ ILC2s in the lymphocyte fractions of healthy controls (n=7) and patients with HDM-AR (n=9) and mugwort-AR (n=8). Each point represents an individual patient sample, and the horizontal bars indicate the mean for each group. Significance of differences was analyzed by the Mann-Whitney U test (**P<0.01).

• 

  Fig. 3 IL-2-, IL-33-, and/or IL-25-induced (A) IL-5 and (B) IL-13 release from the PBMCs of AR patients (n=9). Data are shown as mean±SEM, and significance of differences were analyzed by the Mann-Whitney U test (*P<0.05; **P<0.01).

• 

  Fig. 4 IL-2-, IL-33-, and/or IL-25-induced release of (A) IL-5 and (B) IL-13 from the PBMCs of patients monosensitized to HDM (HDM-AR, n=5) or mugwort (mugwort-AR, n=4). Data are shown as mean±SEM, and significance of differences were analyzed by the Mann-Whitney U test (*P<0.05).

• 

  Fig. 5 IL-2-, IL-33-, and/or IL-25-induced release of (A) IL-5 and (B) IL-13 from Lineage- and Lineage+ cells separated from the PBMCs of patients with HMD-AR. Data are shown as mean±SEM, and significance of differences were analyzed by the Mann-Whitney U test (*P<0.05).

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