| Summary: | 碩士 === 國立中興大學 === 生物化學研究所 === 90 === Most asthma is found in association with immediate hypersensitivity to inhaled allergens, the house dust mite is one of the airway hyper-responsiveness (AHR) allergens. The recently researches reported that the excessively allergenic challenge would cause the hyposensitivity, which will inhibit the Th cell activity and the IgE production, whereas increase the IgG concentration. To examine and develop the novel allergen immunization approach in altering allergenic response, transgenic mice carrying mite’s allergenic genes were built as an animal model. In this study, we have been successfully generated Der p 5 and Der p 2-riched milk in vivo by expressing the Der p 5 and Der p 2 allergen in the mammary gland during lactation which under the control of the promoter of the milk protein, a-lactalbumin. In aLA-derp5i transgenic mice strain, the Der p 5 protein had been found to accumulate in the mammary cell. However, in aLA-derp5t strain that included the aS1 casein signal peptide, the Der p 5 protein had been detected to secrete in secretary tubules by immunofluorescent staining of mammary gland tissue. The lung functional assay to examine the Der p 5 protein biological function, mice sucked the Derp5-containing milk have batter lung capacity after challenged by the allergen. In the second experiment, aLA-derp2t transgenic mice were generated and demonstrated that Der p 2 protein can be secrete in the transgenic milk by Western blot assay. The temporal and spatial expression of derp 2 mRNA in the mammary epithelium cells was regulated with the aLA promoter been demonstrated by RT-PCR at different lactating stage. In the animal test, we also demonstrated that mice sucked the Derp2-containing milk have less inflammation reaction showed in histological assay and ELISA analysis. In the future, the new oral drug immunotherapy could be modulate the IgE antibody response and treated the AHR in atrophic patient.
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