Recombinant Attenuated Salmonella Vaccines for Humans

描述（由申请人提供）：在过去十年中，每年平均有5700万人死亡，其中约1480万人直接死于传染病，另有数百万人死于感染的继发效应（65）。每年因伤寒沙门菌和甲型副伤寒沙门菌、乙型肝炎病毒（HBV）、肺炎链球菌和结核分枝杆菌（Mtb）感染而死亡的人数差别很大，但可能占总死亡人数的30%左右，与这些疾病相关的严重发病率造成的成本甚至更高（232）。相信改善健康、营养和经济福利（后者取决于前两者）是提高全球生活质量的最佳手段，从而减少导致战争和恐怖行为的条件，我们根据我们最近在使用重组减毒沙门氏菌疫苗（RASV）方面的技术发展，提出了一项疫苗开发计划。我们的具体目标是：(i)通过增加膜囊泡的产生和释放以及通过改变RASV细胞粘附属性来增强对RASV表达的保护性肺炎球菌抗原（作为模型抗原）的粘膜和全身抗体反应的诱导；（ii）通过确保不能在体内（特别是在鸟类和动物的肠道中）持续存在和/或完全溶解来增强对活的RASV菌株的生物遏制。（iii）设计和构建A型副伤寒沙门氏菌作为抗原和DNA疫苗递送载体，具有多种遗传特异性特征，以确保用于对婴儿和成人进行免疫以预防肠热病和其他传染病时的安全性和有效性；构建和评估重组减毒伤寒沙门氏菌和甲型副伤寒沙门氏菌疫苗，通过II型和III型分泌传递保护性抗原和/或通过体内调节的延迟裂解(裂解发生在细胞室中，以增强MHC I类或II类的抗原呈递，并在细胞质室中调节延迟裂解以释放针对核靶向优化的DNA疫苗，从而合成和修饰Mtb免疫个体的保护性抗原。我们还将添加我们的主文件（向FDA提交），准备和充分表征候选疫苗主种子的稳定性和安全性，准备和提交IRB批准的协议，提交获得ind所需的信息，并执行任何其他必要的工作，以安排最佳候选疫苗在人类志愿者中进行临床评估。公共卫生相关性：该研究计划将进一步发展和完善设计和构建重组减毒沙门氏菌活疫苗（RASV）载体的技术，在无针口服接种后提供保护性抗原和/或DNA疫苗，以刺激粘膜、全身和细胞免疫，抵抗细菌、病毒和寄生虫传染病病原体。更具体地说，我们将开发一种安全、有效的甲型副伤寒沙门氏菌疫苗载体，用于新生儿和婴儿的口服免疫。我们将使用这种副伤寒沙门氏菌载体和伤寒沙门氏菌载体构建rasv，以防止结核分枝杆菌的传播和感染。

DESCRIPTION (provided by applicant): Of the average 57 million annual deaths over the past ten years, about 14.8 million are directly due to infectious diseases with millions more due to secondary effects of infections (65). The numbers of annual deaths due to infections by Salmonella Typhi and S. Paratyphi A, hepatitis B virus (HBV), Streptococcus pneumoniae and Mycobacterium tuberculosis (Mtb) vary widely but may account for about 30 percent of the total deaths with an even greater cost being due to the severe morbidity associated with these diseases (232). In the belief that improving health, nutrition and economic well being (the latter dependent on the first two) provides the best means to enhance the quality of life globally and thus reduces conditions that result in warlike and terrorist behavior, we propose a vaccine development program based on our recent technical developments in using recombinant attenuated Salmonella vaccines (RASV). Our Specific Aims are (i) to enhance induction of mucosal and systemic antibody responses to RASV-expressed protective pneumococcal antigens (as a model antigen) by increased production and release of membrane vesicles and by altering RASV cell adherence attributes, (ii) to enhance biological containment of live RASV strains by ensuring inability to persist and/or to completely lyse in vivo and especially in the intestinal tracts of birds and animals, (iii) to design and construct S. Paratyphi A as an antigen and DNA vaccine delivery vector with a diversity of genetically-specified features to ensure safety and efficacy when used to immunize infants and adults to protect against enteric fever and additional infectious diseases, and (iv) to design, construct and evaluate recombinant attenuated S. Typhi and S. Paratyphi A vaccines to prevent infections by Mtb using delivery of protective antigens by Type II and III secretion and/or by regulated delayed lysis in vivo with lysis occurring in cell compartments to enhance antigen presentation either by MHC Class I or Class II and by regulated delayed lysis in vivo in the cytoplasmic compartment to release a DNA vaccine optimized for nuclear targeting leading to synthesis and modification of protective antigens by the immunized individual. We will also add to our Master File (filed with FDA), prepare and fully characterize candidate vaccine Master Seeds for stability and safety, prepare and submit protocols for IRB approvals, submit information necessary to obtain INDs, and perform any other work needed to arrange that the best candidate vaccines be clinically evaluated in human volunteers. PUBLIC HEALTH RELEVANCE: The research proposed will further develop and refine technologies for design and construction of live recombinant attenuated Salmonella vaccine (RASV) vectors to deliver after needle-free oral inoculation protective antigens and/or DNA vaccines to stimulate mucosal, systemic and cellular immunities against bacterial, viral and parasite infectious disease agents. More specifically, we will develop a safe, efficacious S. Paratyphi A vaccine vector for oral immunization of newborns and infants. We will use this S. Paratyphi A vector in addition to an S. Typhi vector to construct RASVs to prevent transmission and infection by Mycobacterium tuberculosis.