NOVEL APPROACHES TO DENDRITIC CELL STIMULATION

SIV-smm-pbj-14株是一种独特的灵长类慢病毒， 含有基于免疫受体的酪氨酸激活的Nef蛋白 基序（ITAM）。这种ITAM通过与ZAP-70相互作用激活T细胞 酪氨酸激酶是T细胞受体复合物（TCR）的关键组分。的 该建议的假设是，该Nef-ITAM可以激活细胞内 信号通路，从而增强免疫刺激， 抗原呈递两个具体目标解决了这一假设。要求1 描述了各种质粒构建体和腺病毒载体的构建 设计成与靶抗原一起表达几种形式的Nef-ITAM， 它是HIV-1包膜糖蛋白。目标2评估以下方面的潜在作用： Nef-ITAM对人和小鼠培养物中抗原提呈功能影响 用腺病毒载体感染树突状细胞。额外的体内 用腺病毒载体在小鼠中进行的实验测试了 Nef-ITAM对包膜糖蛋白免疫应答的影响。

The SIV-smm-pbj-14 strain is a unique primate lentivirus that encodes a Nef protein containing an immunoreceptor-based tyrosine activating motif (ITAM). This ITAM activates T-cells through interaction with ZAP-70 tyrosine kinase, a key component of the T-cell receptor complex (TCR). The hypothesis for this proposal is that this Nef-ITAM may activate intracellular signal pathways in dendritic cells, and thereby enhance immune stimulation and antigen presentation. Two Specific Aims address this hypothesis. Aim 1 describes the construction of various plasmid constructs and adenovirus vectors designed to express several forms of Nef-ITAM together with the target antigen, which is the HIV-1 envelope glycoprotein. Aim 2 assesses the potential role of the Nef-ITAM on antigen presentation functions in cultures of human and mouse dendritic cells infected with adenovirus vectors. Additional in vivo experiments in mice with the adenovirus vectors test the effects of the Nef-ITAM on immune responses to the envelope glycoprotein.