Allergy Asthma Immunol Res.  2017 Jan;9(1):61-69. 10.4168/aair.2017.9.1.61.

Inhibition of Allergic Response by Intranasal Selective NF-κB Decoy Oligodeoxynucleotides in a Murine Model of Allergic Rhinitis

• Wee JH
• Zhang YL
• Rhee CS
• Kim DY

• KMID: 2355895
• DOI: http://doi.org/10.4168/aair.2017.9.1.61

Abstract

PURPOSE
It remains unknown whether local inhibition of Nuclear factor-kappa B (NF-κB) could have therapeutic value in the treatment of allergic rhinitis (AR). This study aimed to evaluate the effect of selective NF-κB inhibition using NF-κB decoy oligodeoxynucleotides (ODNs) for the local treatment of AR in ovalbumin (OVA)-sensitized wild-type mice.
METHODS
BALB/c mice were sensitized with OVA and alum, and then challenged intranasally with OVA. NF-κB decoy ODNs were given intranasally to the treatment group, and NF-κB scrambled ODNs were given to the sham treatment group. Allergic symptom scores, eosinophil infiltration, cytokine levels in the nasal mucosa, nasal lavage fluid, and spleen cell culture, serum total and OVA-specific immunoglobulins, as well as intercellular adhesion molecure-1 (ICAM-1) in the nasal mucosa, were analyzed.
RESULTS
NF-κB decoy ODNs significantly reduced allergic symptoms and eosinophil infiltration in the nasal mucosa. They also suppressed serum levels of total IgE, OVA-specific IgE, and IgG1. IL-5 and TNF-α levels and the expression of ICAM-1 were decreased in the nasal mucosa of the treatment group compared to the positive control and sham treatment groups. In addition, IL-6 levels were significantly decreased in the nasal lavage fluid of the treatment group. Furthermore, NF-κB decoy ODNs significantly reduced expression of the systemic Th2 cytokines, IL-4 and IL-5 in spleen cell culture.
CONCLUSIONS
This study demonstrates for the first time that local NF-κB inhibition using NF-κB decoy ODNs suppressed the allergic response in a murine AR model. This shows the therapeutic potential of local NF-κB inhibition in the control of AR.

Keyword

Allergic rhinitis; anti-allergic agents; NF-kappa B; NF-kappa B decoy; oligodeoxyribonucleotides; immunoglobulin E

MeSH Terms

Animals
Anti-Allergic Agents
Cell Culture Techniques
Cytokines
Eosinophils
Immunoglobulin E
Immunoglobulin G
Immunoglobulins
Intercellular Adhesion Molecule-1
Interleukin-4
Interleukin-5
Interleukin-6
Mice
Nasal Lavage Fluid
Nasal Mucosa
NF-kappa B
Oligodeoxyribonucleotides*
Ovalbumin
Ovum
Placebos
Rhinitis, Allergic*
Spleen
Anti-Allergic Agents
Cytokines
Immunoglobulin E
Immunoglobulin G
Immunoglobulins
Intercellular Adhesion Molecule-1
Interleukin-4
Interleukin-5
Interleukin-6
NF-kappa B
Oligodeoxyribonucleotides
Ovalbumin
Placebos

Figure

• 

  Fig. 1 Experimental protocol. BALB/c mice were sensitized with intraperitoneal injection of ovalbumine (OVA) and alum on days 0, 7, and 14. Intranasal challenges were administered via daily OVA instillation from days 20 to 26. The control mice (group A) were intraperitoneally injected and intranasally chal-lenged with phosphate-buffered saline (PBS) instead of OVA on the same schedule. In addition to sensitization and challenge, mice of group C were treated with nasal instillation of the nuclear factor-kappa B (NF-κB) decoy oligodeoxynucleotides (ODNs) and those of group D were sham treated with NF-κB scrambled ODNs on days 20, 22, 24, and 26 (6 hours before nasal challenge). On day 27, after intranasal allergen provocation with 100 µg of OVA, the frequencies of sneezing and nasal rubbing behaviors were recorded during 15 minutes. Mice were killed 24 hours after the final OVA challenge.

• 

  Fig. 2 Symptom scores. Local treatment of NF-κB inhibitor (Group C) suppressed allergic symptoms. Sneezing symptom score (A) and nasal rubbing symptom score (B) of each group. Data are expressed as mean±standard error mean (SEM). *P<0.05.

• 

  Fig. 3 Local treatment of NF-κB inhibitor (Group C) suppressed eosinophil infiltration in the nasal mucosa. Sirius red staining (×400 magnification) of each group (A). Photographs of representative nasal mucosa in each group. Eosinophil count of the septal mucosa of each group (B, n=6). Data are expressed as mean±standard error mean (SEM). *P<0.05.

• 

  Fig. 4 Expression of IL-4 (A), IL-5 (B), IL-6 (C), TNF-α (D), and IFN-γ (E) in the nasal mucosa by real time PCR. The transcriptional levels of IL-5 and TNF-α were decreased in treatment group C than positive control group B. Data are expressed as mean±standard error mean (SEM). *P<0.05.

• 

  Fig. 5 Concentrations of IL-6 in nasal lavage fluid of each group by ELISA. The levels of IL-6 were significantly decreased in treatment group C than positive control group B. Data are expressed as mean±standard error mean (SEM). *P<0.05.

• 

  Fig. 6 Concentrations of IL-4 (A), IL-5 (B), IL-10 (C), and IFN-γ (D) in spleen cell culture of each group by ELISA. Systemic Th2 cytokines (IL-4 and IL-5) were significantly decreased in treatment group C than in positive control group B. However, systemic regulatory cytokine (IL-10) or Th1 cytokine (IFN-γ) did not change in treatment group C. Data are expressed as mean±standard error mean (SEM). *P<0.05.

• 

  Fig. 7 Serum immunoglobulin levels in each group. Local treatment with NF-κB decoy ODNs (Group C) significantly suppressed levels of serum total IgE (A), OVA-specific IgE (B), and IgG1 (C). The serum levels of OVA-specific IgG2a (D) were increased. Data are expressed as mean±standard error mean (SEM). *P<0.05.

• 

  Fig. 8 Local treatment with NF-κB decoy ODNs (Group C) significantly reduced the expression of ICAM-1 protein in the nasal mucosa by Western blot. Immunoblot of ICAM-1 and β-actin in each group (A). Photographs of representative sample in each group. Quantitative expression level of ICAM-1/β-actin in the nasal mucosa of each group (B, n=6). Data are expressed as mean±standard error mean (SEM). *P<0.05.

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