The role of lung epithelium in asthma inflammation

• Main Authors: Wan-Ning Wang, 王琬甯
• Other Authors: Yi-Ling Ye
• Format: Others
• Language: zh-TW
• Published: 2005
• Online Access: http://ndltd.ncl.edu.tw/handle/93932263558071909589

碩士 === 中華醫事學院 === 生物科技研究所 === 92 === Asthma is a Th2 type inflammatory immune response. Cytokines and cells belonging to innate immunity contribute to the Th2 response. In asthma inflammatory response, the chemokines secreted by lung epithelium after antigen stimulation will subsequently influence the direction of immune response. We use different antigens co-cultured with lung cells of C57BL/6 mice to develop an in vitro system in order to mimic the in vivo condition. Then analyze the expression of chemokines by ELISA or RT-PCR assay. According to the recent study about the role of lipopolysaccharide in the establishment of OVA asthma animal model, we also measure the content of endotoxin in our antigens by LAL assay. There are many kinds of cells exist in the lung epithelium. We use immuno-histochemistry to analyze it. Furthermore, mice were stimulated with different antigens by intra nasal administration. After 7 days stimulation, the bronchial alveolar larvage fluids (BALFs) was collected then analyze the kinds of cells and chemokines secretion. We found that OVA, LPS, Der pI and Der pII can stimulate lung cells to secrete high level of RANTES, but BSA could not do it. OVA also can induce the higher secretion of eotaxin than BSA and the more serious airway inflammation than BSA in our in vivo experiment. Eodotoxin could not influence our result because the level exist in our experiment is lower than 10-19ug/mL(0.173EU/mL) LPS. Fibroblasts were the major cell type in our primary lung cell culture. We also found epithelial cells exist in it. In our study, we establish an in vitro primary lung cell culture system and evaluate the secretion ability after different antigen stimulation. We also use in vivo assay to prove the in vitro results. It verifies that lung cells could secrete chemokines and contribute to the subsequent inflammation after proper antigen stimulation. We hope that this primary culture system can provide us to evaluate the first response after allergens enter into our human body. It is also interesting that different antigen induce different chemokine expression pattern. Furthermore studies are needed to answer these questions.