Analysis of bone-marrow derived fibroblasts involved in asthmatic airway remodeling and its interventional control of function

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Analysis of the asthma model with bone marrow transferred chimeric miceMethod : We made the murine asthma model with bone marrow transferred chimeric C57BL/6 mice. At two weeks after sensitization with Ovalbumin (OVA), the mice were exposed to OVA every other day during one month. Twenty four hours later from final exposure with OVA, we collected blood and broncho-alveolar lavage fluid (BAL) and obtained the lung tissue from these mice on 7^<th> day, 14^<th> day and 28^<th> day. In addition, we divided these mice into two groups. One group of the mice was administered intraperitoneally with G-CSF and the other group with saline as the control.Estimation of histological changes was performed. The pulmonary tissue was fixed with formaldehyde and were embedded in paraffin. On the tissue sections, we estimated infiltrated inflammatory cells into airway wall as a score semi-quantitatively with Hematoxylin-Eosin staining. We identified GFP-expressing cells in bronchial tissues under chemiluminescence microscopy and also GFP positive cells with immunohistochemistry using ant-GFP antibody.Results : On 7^<th> day after OVA exposure, eosinophils as major cells, lymphocytes and monocytes were observed in peribronchial and alveolar tissue in the chimeric mice. On 14^<th> and 28^<th> day, thickening of epithelial basement membrane and deposition of collagen in submucosal layer of bronchi became prominent. Some GFP-expressing cells were spindle-like shape under epithelium and assumed fibroblast like cells. They were also confirmed by immunohistochemistry.GFP-expressing cells increased more in the lung of the mice with G-CSF treatment than control, and collagen deposition estimated with Elastica-Masson staining decreased in G-CSF treated mice compared to control.Conclusion : Bone marrow derived cells were recruited to peribronchial tissue with allergic inflammation and might play a regulatory role in collagen deposition in the airway.