Study On Expression And Role Of Th1 Type Transcription Factor T-bet In Nasal Mucosa Epithelial Cells By Recombined Adenovirus-Mediated

The transcription factor T-bet promotes the differentiation of inflammatory Th1 helper cells. It was revealed by Szabo in 2000. Usually, T-bet expression in lymphoid cells is regulated by cytoplasmic signaling through Janus kinase phosphorylation, nuclear signaling using signal transducers and activators of transcription(Stat) family proteins, and autocrine/paracrine feedback involving interferon-γ(IFNγ). Interestingly, in our test, T-bet was shown to be present in epithelial cells of the humen nasal mucosa(hNMECs) in vitro. Then regulation of T-bet expression was modulated by cytokines interleukin-15(IL-15). This mechanisms of T-bet regulation in epithelia differ from those in conventional immune cells. Although there is no autocrine IFNγfeedback loop in hNMECs, an IL-15/T-bet positive feedback loop may exist. It is important to Allergic Rhinitis(AR) that T-bet activates IFNγand represses IL-4 production in developing Th2 cells, and redirects effector Th2 cells into the Th1 pathway to convert polarized Th2 cells into Th1 cells. A novel target is clear on treatment to AR.We constructed adenovirus with T-bet gene, named rAdT-bet, which tite was 10^5.4TCID50/ml. To further test the tissue-specific expression of T-bet and identify whether T-bet is responsible for the production of cytokine in epithelial cells in vitro, recombined adenovirus gene-mediated transfer of T-bet into hNMECs in vitro was performed. IL-15,IL-4,IFNγproduction was measured by ELISA after hNMECs were infected with rAdT-bet 24h, 48h and 72h. On 24 hours, the transduced level of T-bet mRNA increased 71%(p<0.005) versus the control whithout transduction with rAdT-bet, 48h, increased 74%(p<0.005), 72h, increased 67%(p<0.005). The T-bet protein was measured showing the increase on 21.3%((p<0.005)(the transduction versus the control). On 24h, a dramatic increase was seen in IL-15 on 86%(p<0.005),48h 68%(p<0.005),72h 73%(p<0.005)。Whereas, IL-4 and IFNγproduction remained unchanged.Then during the 12h and 24h exposure to IL-15,IL-4,IFNγ, hNMECs T-bet mRNA levels were measured. The group exposured to IL-15, 12h T-bet mRNA levels increased 50.2%(p<0.005) compared to the control without any stimulation,24h, 66.3%(p<0.005)。The hNMECs had no responsibility to IL-4 and IFNγ.So we concluded that it was high effect the transduction of recombined adenovirus to hNMECs. T-bet expression was shown in hNMECs under the regulation of cytokine in vitro. The expression levels of T-bet in hNMECs was responsible for IL-15 production. In hNMECs, IL-15/T-bet positive feedback loop may exist.In brief, in hNMECS, the T-bet/IL-15 positive feedback loop has an marked advantage to the study on molecule mechanisms and gene treatment of AR. the recombined adenovirus is a high effect vector using to infect hNMECs, but the implications of rAdT-bet played a role in the Mucosa Immune System will be discussed by zoopery.