Immunization Against Pseudomonas aeruginosa by Modified Ad Vectors

描述（由申请人提供）：该项目的目标是利用一种新型腺病毒（Ad）为基础的基因转移载体开发一种抗铜绿假单胞菌疫苗，该载体：编码铜绿假单胞菌外膜蛋白F (OprF)；在纤维中加入RGD序列，通过3.5整合素增强向体内树突状细胞的传递；并将Epi8（一个保守的、高度免疫原性的OprF表位）整合到六联体外环中。目的1侧重于将这一策略用于基于ad5的疫苗（Ad5cuOprF.RGD.Epi8）。我们预计它会引起高水平的抗p。并且衣壳上的Epi8表位将允许增强，即使在第一次免疫诱导的Ad免疫存在的情况下。在完成毒理学试验并获得监管部门批准后，我们将对Ad5cuOprF.RGD进行临床试验。正常成人志愿者的Epi8。在目标2中，我们将使用AdC7cuOprF.RGD进行研究。Epi8，第二代抗p。与Ad5CuOprF.RGD相似的铜绿球菌疫苗。但基于非人类灵长类动物AdC7血清型，即使存在预先存在的抗ad5免疫，这种血清型也应该有效。该项目的具体目的是：目的1 -测试Ad5cuOprF.RGD的管理是否有效的假设。Epi8是一种人类Ad血清5型腺病毒，编码铜绿假单胞菌抗原OprF，并在纤维中进一步修饰树突状细胞靶向基序RGD和OprF Epi8表位在六元上，是安全的，并将在健康个体的血液和肺上皮内层液中产生强大的抗假单胞菌调理抗体；目的2 -检验有效反p的假设。AdC7cuOprF.RGD可在抗ad5免疫的背景下实现铜绿菌免疫。Epi8，与Ad5cuOprF.RGD相同的载体。但基于AdC7，一种人类没有免疫力的非人类灵长类腺病毒。呼吸道感染铜绿假单胞菌是囊性纤维化患者发病和死亡的主要原因。目前还没有上市的预防铜绿假单胞菌感染的疫苗，因此成功的疫苗策略将对CF患者产生深远的影响。

DESCRIPTION (provided by applicant): The goal of this project is to develop an anti-Pseudomonas aeruginosa vaccine using a novel adenovirus (Ad)-based gene transfer vector that: codes for the P. aeruginosa outer membrane protein F (OprF); has been enhanced to incorporate an RGD sequence in the fiber to enhance delivery to dendritic cells in vivo via ?v?3.5 integrins; and incorporates Epi8, a conserved, highly immunogenic OprF epitope, into the hexon outer loop. Aim 1 focuses on using this strategy with an Ad5-based vaccine (Ad5cuOprF.RGD.Epi8). We expect that it will elicit high levels of anti-P. aeruginosa antibodies in blood and lung, and that the Epi8 epitope on the capsid will allow boosting, even in the presence of immunity against the Ad induced by the 1st immunization. After completing the toxicology testing, and gaining regulatory approval, we will carry out a clinical trial with Ad5cuOprF.RGD.Epi8 in normal adult volunteers. In aim 2, we will carry out studies with AdC7cuOprF.RGD.Epi8, a 2nd generation anti-P. aeruginosa vaccine that is similar to Ad5CuOprF.RGD.Epi8, but based on the non-human primate AdC7 serotype, a serotype that should be effective even in the presence of pre-existing anti-Ad5 immunity. The specific aims of the project are: Aim 1 - To test the hypothesis that administration of Ad5cuOprF.RGD.Epi8, a human Ad serotype 5 adenovirus that codes for the P. aeruginosa antigen OprF and is further modified with a dendritic cell targeting motif RGD in the fiber and the OprF Epi8 epitope in the hexon, is safe and will evoke robust anti-Pseudomonas opsonizing antibodies in blood and lung epithelial lining fluid of healthy individuals; Aim 2 - To examine the hypothesis that effective anti-P. aeruginosa immunity can be achieved in the context of anti-Ad5 immunity with AdC7cuOprF.RGD.Epi8, a vector identical to Ad5cuOprF.RGD.Epi8 but based on AdC7, a non-human primate adenovirus against which humans do not have immunity. Infections of the respiratory tract with Pseudomonas aeruginosa is the major cause of morbidity and mortality in individuals with cystic fibrosis. A marketed vaccine for prevention of infection with Pseudomonas aeruginosa is not available, therefore a successful vaccine strategy would have profound effects on individuals with CF.