Allergy Asthma Immunol Res.  2017 Nov;9(6):540-549. 10.4168/aair.2017.9.6.540.

Mouse Model of IL-17-Dominant Rhinitis Using Polyinosinic-Polycytidylic Acid

• Bae JS
• Kim EH
• Kim JH
• Mo JH

• KMID: 2421665
• DOI: http://doi.org/10.4168/aair.2017.9.6.540

Abstract

Interleukin (IL)-17 plays an important role in rhinitis and the level thereof correlates with the severity of disease. However, no mouse model for IL-17-dominant rhinitis has yet been developed. Our objective was to establish a mouse model of IL-17-dominant rhinitis via intranasal application of polyinosinic-polycytidylic acid (abbreviated as Poly(I:C)). Mice were divided into 6 groups (n=8 for each group); 1) 1 negative control group, 2) 1 positive control group (OVA/alum model), 3) 2 Poly(I:C) groups (10 or 100 µg), and 4) 2 OVA/Poly(I:C) groups (10 or 100 µg). The positive control group was treated with the conventional OVA/alum protocol. In the Poly(I:C) and OVA/Poly(I:C) groups, phosphate-buffered saline or an OVA solution plus Poly(I:C) were administered. The OVA/Poly(I:C) groups exhibited significantly greater neutrophil infiltration and increased IL-17/interferon γ expression compared with the other groups. However, the levels of total immunoglobulin E (IgE), OVA-specific IgE, eosinophil infiltration, IL-4, IL-5, IL-6, and IL-10 were significantly lower in the OVA/Poly(I:C) groups than in mice subjected to conventional Th2-dominant OVA/alum treatment (the positive control group). IL-17 and neutrophil measurement, chemokine (C-X-C motif) ligand 1 immunohistochemistry, and confocal microscopy revealed increased numbers of IL-17-secreting cells in the nasal mucosa of the OVA/Poly(I:C) groups, which included natural killer cells, CD4 T cells, and neutrophils. In conclusion, we developed a mouse model of IL-17-dominant rhinitis using OVA together with Poly(I:C). This model will be useful in research on neutrophil- or IL-17-dominant rhinitis.

Keyword

Rhinitis; mouse model; polyinosinic-polycytidylic acid; IL-17; neutrophil

MeSH Terms

Animals
Chemokine CXCL1
Eosinophils
Immunoglobulin E
Immunoglobulins
Immunohistochemistry
Interleukin-10
Interleukin-17
Interleukin-4
Interleukin-5
Interleukin-6
Interleukins
Killer Cells, Natural
Mice*
Microscopy, Confocal
Nasal Mucosa
Neutrophil Infiltration
Neutrophils
Ovum
Poly I-C*
Rhinitis*
T-Lymphocytes
Chemokine CXCL1
Immunoglobulin E
Immunoglobulins
Interleukin-10
Interleukin-17
Interleukin-4
Interleukin-5
Interleukin-6
Interleukins
Poly I-C

Figure

• 

  Fig. 1 (A) Protocol for the induction of AR with OVA and Poly(I:C) in mice. The mice were divided into 6 groups: 1) 1 negative control group, 2) 1 positive control group (OVA/alum model), 3) 2 Poly(I:C) groups (10 or 100 µg), and 4) 2 OVA/Poly(I:C) groups (10 or 100 µg). The positive control group was injected with OVA and alum mixtures intraperitoneally (i.p.) on days 0, 7, and 14 for sensitization. In Poly(I:C) or OVA/Poly(I:C) groups, PBS/Poly(I:C) or OVA/Poly(I:C) mixtures were treated intranasally (i.n.) on days 0, 1, 2, 7, and 14. For challenge, PBS or OVA were administered intranasally on days 14 to 21. (B) Symptom score of nasal rubbing and sneezing for 15 minutes. (C) The total and OVA-specific immunoglobulin (IgE, IgG1, and IgG2a) levels in the serum. AR, allergic rhinitis; OVA, ovalbumin; Poly(I:C), polyinosinic-polycytidylic acid; PBS, phosphate-buffered saline; Ig, immunoglobulin. *P<0.05; **P<0.01; ***P<0.001.

• 

  Fig. 2 Histological staining for inflammatory and IL-17A-producing cells in the nasal mucosa. (A) Eosinophil counts in the nasal mucosa (×400 magnification, Sirius red staining). (B) Neutrophil counts in the nasal mucosa (×400 magnification, IHC with NIMP-R14 antibody), and (C) IL-17A producing cell counts in nasal tissues (×400, IHC with IL-17A antibody). IL, interleukin; IHC, immunohistochemistry. *P<0.05; **P<0.01.

• 

  Fig. 3 (A) Local cytokine production in the nasal mucosa, mRNA expression (IL-4, IL-5, IL-6, IL-10, IL-17, and IFN-γ). (B) Systemic cytokine production from spleen cell culture. IL, interleukin; IFN, interferon. *P<0.05; **P<0.01; ***P<0.001.

• 

  Fig. 4 IHC of (A) NKp46 and (B) CXCL1 (both ×400 magnification). IHC, immunohistochemistry; CXCL1, chemokine ligand 1. *P<0.05; **P<0.01; ***P<0.001.

• 

  Fig. 5 Immunofluorescent double staining with (A) IL-17 (red)/NKp46 (green), (B) IL-17 (green)/CD4 (red), (C) IL-17 (green)/NIMP-R14 (red), and (D) IL-17 (red)/MBP (green) in the nasal mucosa. Blue indicates DAPI nuclear counterstaining. Orange colored portion in merged image means double positive cells. IL, interleukin; MBP, major basic protein; DAPI, 4′,6-diamidino-2-phenylindole.

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