Allergy Asthma Immunol Res.  2011 Jul;3(3):199-204. 10.4168/aair.2011.3.3.199.

The Role of Plasmacytoid and Myeloid Dendritic Cells in Induction of Asthma in a Mouse Model and the Effect of a TLR9 Agonist on Dendritic Cells

• Mo JH
• Chung YJ
• Hayashi T
• Lee J
• Raz E

• KMID: 1803326
• DOI: http://doi.org/10.4168/aair.2011.3.3.199

Abstract

PURPOSE
To determine the role of plasmacytoid dendritic cells (pDC) and myeloid dendritic cells (mDC) in priming effector T cells to induce allergy, and to evaluate the effect of immunostimulatory sequences (ISS, TLR9 agonist) on dendritic cells.
METHODS
Cultured mDC and pDC with/without ISS were injected intratracheally into sensitized Balb/C mice. Mice were sacrificed, and then pulmonary function tests, bronchoalveolar lavage (BAL), cell counts, and cytokine levels were evaluated. Migration of dendritic cells was also evaluated after ISS administration.
RESULTS
In mice injected with mDC, airway hyperresponsiveness, eosinophil counts, and Th2 cytokine levels in BAL increased with increasing numbers of mDC injected. However, in mice injected with pDC, none of these changed, suggesting poor priming of T cells by pDC. In addition, mDC pulsed with ISS inhibited asthmatic reactions, and ISS administration inhibited migration of DC to the lung.
CONCLUSIONS
We suggest that pDC played a limited role in priming T cells in this asthma model and that mDC played a major role in inducing asthma. In addition, ISS inhibited migration of DC to the lung.

Keyword

Plasmacytoid dendritic cell; myeloid dendritic cell; allergy; immunostimulatory sequence; TLR9; migration

MeSH Terms

Animals
Asthma
Bronchoalveolar Lavage
Cell Count
Dendritic Cells
Eosinophils
Hypersensitivity
Lung
Mice
Respiratory Function Tests
T-Lymphocytes

Figure

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  Fig. 1 Myeloid dendritic cells (DC) induced an asthmatic reaction in myeloid dendritic cells (mDC) in a number-dependent manner. (A) Schematic illustration of the experimental protocol. Different numbers of myeloid DC were administered intratracheally to Balb/c mice after ovalbumin/alum sensitization. Five mice were used in each group and this experiment was performed in triplicate. (B) Airway hyperresponsiveness to methacholine was evaluated. mDC increased airway hyperresponsiveness in a number-dependent manner. (C) Eosinophil counts in bronchoalveolar lavage (BAL) fluid also depended on the number of mDC administered. (D) IL-4 and IL-5 levels increased with increasing numbers of mDC in BAL fluid. Interferon-γ decreased in group 3. Gp, group; IFN- γ, interferon-γ; MCh: methacholine.

• 

  Fig. 2 Isolation of plasmacytoid dendritic cells (DC) and the effect of plasmacytoid dendritic cells (pDC) with or without an immunostimulatory sequence (ISS) on asthma induction. (A) Schematic illustration of the experimental protocol. Myeloid dendritic cells (mDC) or pDC (106) with or without ISS stimulation were administered intratracheally to Balb/c mice after ovalbumin (OVA)/alum sensitization. Five mice were used in each group and this experiment was performed in duplicate. (B) Characterization of pDC subsets after isolation using anti-CD11c and anti-B220 MACS antibodies. Isolated cells exhibited 94.4% purity. (C, D) Airway hyperresponsiveness curve showing that pDC with or without OVA pulsing did not induce airway hyperresponsiveness compared to mDC without OVA (P>0.05). mDC pulsed with OVA increased airway hyperresponsiveness, eosinophil counts, and IL-4 and IL-5 levels in BAL, the effect of which was abolished by administration of mDC pulsed with ISS and OVA.

• 

  Fig. 3 Immunostimulatory sequence (ISS) inhibits myeloid dendritic cells (mDC) migration into the lung of mice in vivo. (A) Schematic illustration of the experimental protocol. Mice were sensitized with ovalbumin (OVA)/alum and ISS was administered intraperitoneally on day 19. Intranasal OVA challenge was performed on days 20 and 25, and this experiment was performed in duplicate. (B) CD11c+CD11b+ cell counts increased from 1.84% to 15.80% after OVA challenge and decreased to 9.02% in the ISS administration group. In the case of CD11c+B220+ cells, counts increased from 0.60% to 1.08% after OVA challenge, and then decreased to 0.69% after ISS administration. (C) ISS administration significantly lowered eosinophil counts and Th2 cytokine (IL-4 and IL-5) levels in BAL fluid (all, P<0.01).

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