CD4+ T cells play a critical role in the development of allergic inflammation in several target organs. Various adhesion molecules are involved in the local recruitment of T cells and other inflammatory cells. We investigated the differential contribution of adhesion molecules to T helper 1 (Th1) and Th2 cell-mediated allergic lung and bowel inflammation by employing their neutralizing antibodies. BALB/c mice transferred with in vitro-differentiated antigen-specific Th1 and Th2 cells were intratracheally or intrarectally challenged with a relevant antigen. Infiltration of infused T cells occurred, along with the accumulation of neutrophils and eosinophils in the lungs of Th1 and Th2 cell-transferred recipients, respectively. Th1-mediated neutrophil and Th2-mediated eosinophil accumulation in the large intestine, which occurred after intrarectal challenge with the antigen, was indicated by the significant elevation of myeloperoxidase (MPO) and eosinophil peroxidase (EPO) activity. Blocking experiments with neutralizing antibodies indicated that intercellular cell adhesion molecule (ICAM)-1; vascular cell adhesion molecule (VCAM)-1; and αL, β2, and β7 integrins participate in the accumulation of Th2 cells and eosinophils in the lungs. In contrast, the migration of Th1 cells and neutrophils was diminished by blockage of αL/β2-integrin and ICAM-1, respectively. Mucosal addressin cell adhesion molecule (MadCAM)-1, vascular cell adhesion molecule (VCAM)-1, α4, β1, and β7 contributed to Th1-mediated neutrophilic inflammation in the bowel, though only MadCAM-1, α4, αL, and β2 were involved in Th2-mediated eosinophilic inflammation. We conclude that distinct sets of adhesion molecules are involved in Th1- and Th2-mediated allergic lung and bowel inflammation.
The eosinophil is deeply associated with the pathogenesis of bronchial asthma and other allergic diseases. We recently identified a novel eosinophil-specific cell surface molecule, major facilitator super family domain containing 10 (Mfsd10). A monoclonal antibody (mAb) against Mfsd10 (M2) showed selective binding and neutralizing activities for eosinophils. However, the relative potency of the blockage of Mfsd10 and other eosinophil-specific molecules for the treatment of allergic diseases has not been evaluated. Therefore, in this study, the effects of M2 and an anti-Siglec-F mAb on antigen-immunized and antigen-specific Th2 cell-transferred murine eosinophilic inflammation models were comparatively investigated. Ovalbumin (OVA)-specific Th2 cells were differentiated from naïve CD4+ T cells of DO11.10/RAG-2−/− mice in vitro and cytokine producing activity of the Th2 cells was examined. OVA-immunized and Th2 cell- transferred BALB/c mice were treated with M2 or anti-Siglec-F and challenged with OVA. Then the number of inflammatory cells and the concentration of IL-5 in the bronchoalveolar lavage fluid (BALF) were determined. Antigen-specific Th2 cells produced large amounts of IL-4, IL-5 and IL-13 but not IL-17A or IFN-γ. Administration of M2 significantly suppressed antigen-induced lung eosinophil infiltration both in OVA-immunized and Th2 cell-transferred mice. The potency as well as selectivity of M2 for down-regulating eosinophils was quite similar to that of anti-Siglec-F. Both mAbs did not affect antigen-induced IL-5 production in the lungs. Mfsd10 as well as Siglec-F could be an effective target to treat eosinophil-related disorders including bronchial asthma.
Background: Oral immunotherapy is potentially useful for the treatment of allergic diseases. We previously demonstrated that allergen-induced airway inflammation and immunoglobulin E (IgE) production in mice were suppressed by oral administration of high-dose transgenic (Tg) rice seeds (approximately 50 g/kg/day) expressing a T cell epitope of Dermatophagoides pteronyssinus group 1 allergen (Der p 1). However, this amount of Tg rice seeds was not realistic in our daily life. In this study, allergen-induced airway inflammation and IgE production following oral immunotherapy with a realistic (lowest) dose of Tg rice seeds were investigated. Methods: Mice orally administered with Tg or non-Tg rice seeds at approximately 5 g/kg/day for 1 week were immunized with recombinant Der p 1, and then challenged with the corresponding allergen. The infiltration of inflammatory cells into the airways and the levels of allergen-specific serum IgE were examined. Results: Low-dose oral administration of Tg rice seeds significantly inhibited the allergen-induced infiltration of eosinophils and lymphocytes into the airways, but allergen-specific IgE synthesis was not changed. Conclusions: Low-dose oral immunotherapy with Tg rice seeds could suppress allergen-induced airway inflammation through mechanisms other than the downregulation of IgE synthesis.
Airway accumulation of eosinophils and bronchial hyperresponsiveness (BHR) are prominent features of bronchial asthma, though the contribution of eosinophils to the development of BHR is controversial. Similar to Th2 cell-mediated pathology, Th9 cells, characterized by IL-9-producing activity, have been demonstrated to induce airway eosinophilia and BHR. In this study, we investigated the role of eosinophils in Th9-mediated BHR by employing Th9 cell-transferred murine airway inflammation model. Ovalbumin (OVA)-specific Th2 and Th9 cells were differentiated from CD4+ T cells of DO11.10/RAG-2−/− mice in vitro and cytokine-producing activity of those cells was examined. BALB/c mice were adoptively transferred with Th2 or Th9 cells and challenged with OVA. Then, the number of inflammatory cells in bronchoalveolar lavage fluid and bronchial responsiveness to inhaled methacholine were determined. Both in Th2 and Th9 cell-transferred mice, substantial accumulation of eosinophils in the lungs and BHR were induced by challenge with specific antigen. Nevertheless, an essential and dispensable role of eosinophils in Th2- and Th9-mediated BHR, respectively, was demonstrated by employing eosinophil-deficient mice. The neutralization of IL-9 as well as deficiency of IL-10 in the donor cells did not affect Th9-mediated BHR. In contrast to Th2-mediated and eosinophil-dependent BHR, Th9 could induce BHR independently from eosinophils and its characteristic cytokines, IL-9 and IL-10.
Activated T cells are crucial for the development of allergic diseases. We have recently clarified that L-type amino acid transporter 1 (LAT1) plays a functional role in activated T cells. Here, we comparatively investigated the effect of a LAT1 inhibitor, JPH203, on allergic inflammation induced in multiple organs, such as the skin, lungs, and nose of antigen-specific Th2 cell-transferred mice. The local antigen provocation to those mice evoked tissue-specific eosinophilic inflammation, especially accompanied by bronchial and nasal hyperresponsiveness (BHR and NHR) in the lungs and nose, respectively. Antigen-induced ear thickness, BHR, and NHR were significantly suppressed by the administration of JPH203, though the attenuation of eosinophil accumulation was only seen in the skin and nose. The infiltration of antigen-specific T cells determined in the lungs and nasal-associated lymphoid tissue was not affected by the JPH203 treatment. Activation-induced amino acid incorporation, oxidative phosphorylation, glycolysis, cyclin-related protein expression, and resulting cytokine synthesis in Th2 cells were suppressed by JPH203. JPH203 is potentially effective for treating allergic diseases through attenuating the function of activated T cells. However, the mechanisms may not involve the suppression of eosinophil or T cell infiltration in some target organs.
