Immunization Against Pseudomonas aeruginosa by Modified Ad Vectors

描述（由申请人提供）：本项目的目标是使用新型基于腺病毒（Ad）的基因转移载体开发抗铜绿假单胞菌疫苗，该基因转移载体：编码铜绿假单胞菌外膜蛋白F（OprF）;已被增强以在纤维中掺入RGD序列，以增强通过？v？3.5整合素;并将Epi 8（一种保守的高度免疫原性OprF表位）掺入六邻体外环。目的1集中于将该策略用于基于Ad 5的疫苗（Ad5cuOprF.RGD.Epi8）。我们预期它将在血液和肺中引发高水平的抗铜绿假单胞菌抗体，并且衣壳上的Epi 8表位将允许加强，即使在存在针对由第一次免疫诱导的Ad的免疫的情况下。在完成毒理学测试并获得监管机构批准后，我们将在正常成年志愿者中进行Ad5cuOprF.RGD.Epi8的临床试验。在目标2中，我们将用AdC7cuOprF.RGD.Epi8进行研究，AdC7cuOprF.RGD.Epi8是第二代抗铜绿假单胞菌疫苗，其类似于Ad5CuOprF.RGD.Epi8，但基于非人灵长类动物AdC 7血清型，该血清型即使在预先存在的抗Ad 5免疫力的情况下也应该是有效的。该项目的具体目标是：目的1 -为了验证以下假设：给予Ad5cuOprF.RGD.Epi8，这是一种编码铜绿假单胞菌抗原OprF的人Ad血清型5腺病毒，并进一步用纤维中的树突状细胞靶向基序RGD和六邻体中的OprF Epi 8表位进行修饰，是安全的，并将在健康个体的血液和肺上皮衬里液中引起强有力的抗假单胞菌调理抗体;目的2 -检验在使用AdC7cuOprF.RGD.Epi8（与Ad5cuOprF.RGD.Epi8相同但基于AdC 7的载体）的抗Ad 5免疫的情况下可以实现有效的抗铜绿假单胞菌免疫的假设，一种人类对其没有免疫力的非人类灵长类动物腺病毒。铜绿假单胞菌呼吸道感染是囊性纤维化患者发病和死亡的主要原因。用于预防铜绿假单胞菌感染的市售疫苗不可用，因此成功的疫苗策略将对CF个体产生深远影响。

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.