The Test of Adeno-Associate Viral Vectors Carrying Allergic Modulatory Genes in a Murine Model

Asthma is a complex pulmonary inflammatory disease. Generally, it is believed that the susceptible individuals have an abnormal immune response to inhaled allergens. According to some previous studies, Th2 cells and their cytokines, IL-4, IL-5, and IL-13 play pivotal roles in the pathogenesis of asthma. The Th2 cytokines cause the differentiation, recruitment, and activation of the mast cells and eosinophils in the airway mucosa. Activation of these cells releases high levels of proinflammatory mediators which induce bronchial obstruction, airway hyper-responsiveness, and airway inflammation. We would first establish the adeno-associate virus (AAV) vectors to carry potential anti-inflammatory molecules, such as Clara cell 10 kD protein (CC10) or siRNA specific to eotaxin-1. The therapeutic potential of AAV vectors carrying these 2 gene fragments will be examined in ovalbumin (OVA)-sensitized mice. In addition, since IL-31 has been shown to be important in the pathogenesis of atopic dermatitis and nearly 80% children with atopic dermatitis develop allergic rhinitis or asthma, we would also apply the AAV gene transfer system to examine the role of IL-31 in asthma pathogenesis. IL-31 signals via the heterodimeric receptor complex composed of IL-31 receptor A (IL-31 RA) and oncostatin M receptor (OSMR). Our preliminary results indicated that IL-31 RA was up-regulated in lung epithelium and bronchoalveolar lavage cells from an animal model of OVA-challenged airway hyper-responsiveness. We will clone IL-31 cDNA from activated splenocytes and construct it into an expression vector to make recombinant protein and to generate AAV vector carrying IL-31. We will examine whether the intratracheal injection of IL-31 protein or AAV-IL-31 would increase of the severity of asthma. The airway hyperresponsiveness, cell infiltration in BALF, or higher expression of chemokines will be evaluated. The BALF cell populations in mice expressing IL-31 RA will be identified. Then, we will generate monoclonal antibodies (MoAbs) specific to IL-31 RA or OSMR. Moreover, we will use adeno-associate vectors that carry tissue specific siRNAs that target IL-31 or IL-31 RA. The importance of IL-31 signaling will be confirmed with the application of the MoAbs or AAV carrying siRNA against IL-31 RA into OVA-sensitized mice. The expression of some regulator molecules for Th2 function, such as STAT3 or SOCS3, will be examined with recombinant IL-31, anti-IL-31 RA, or AAV vector viruses. The results will offer a better understanding of the role and functional mechanisms of IL-31 in allergic diseases. The novel therapeutic approaches with those molecules will be developed based on the information obtained from this study.

Project ID:PC10001-0203
External Project ID:NSC98-2320-B182-030-MY3