Recombinant Attenuated Salmonella Vaccines for Humans

描述（申请人提供）：在过去十年中，平均每年5700万人死亡，约1480万人直接死于传染病，另有数百万人死于感染的继发性影响（65）。每年因伤寒沙门氏菌和沙门氏菌感染而死亡的人数。甲型副伤寒、B肝炎病毒（HBV）、肺炎链球菌和结核分枝杆菌（Mtb）的死亡率差异很大，但可能占总死亡人数的30%左右，由于这些疾病的严重发病率，成本更高（232）。我们相信，改善健康、营养和经济福祉（后者依赖于前两者）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，我们提出了一个疫苗开发计划，该计划基于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。我们的具体目标是（i）增强对RASV表达的保护性肺炎球菌抗原的粘膜和全身抗体应答的诱导（作为模型抗原）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，（ii）通过确保不能持续存在来加强对活的RASV菌株的生物遏制，和/或在体内，特别是在鸟类和动物的肠道中完全裂解，（iii）设计和构建S.甲型副伤寒作为一种抗原和DNA疫苗载体，具有多种遗传特异性特征，以确保用于免疫婴儿和成人时的安全性和有效性，以防止肠热病和其他传染病，和（iv）设计，构建和评估重组减毒沙门氏菌。伤寒和沙门氏菌。使用通过II型和III型分泌递送保护性抗原预防Mtb感染的甲型副伤寒疫苗和/或或通过在细胞区室中发生裂解以增强通过MHC I类或II类的抗原呈递的体内受调节的延迟裂解，以及通过在细胞质区室中的体内受调节的延迟裂解以释放针对核靶向而优化的DNA疫苗，从而导致保护性抗原的合成和修饰，免疫个体。我们还将添加到我们的主文件（提交给FDA），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批准的方案，提交获得IND所需的信息，并执行安排最佳候选疫苗在人类志愿者中进行临床评价所需的任何其他工作。公共卫生相关性：拟议的研究将进一步开发和完善设计和构建活重组减毒沙门氏菌疫苗（RASV）载体的技术，以在无针口服接种后提供保护性抗原和/或DNA疫苗，以刺激针对细菌，病毒和寄生虫传染病病原体的粘膜，系统和细胞免疫。更具体地说，我们将开发一种安全有效的S。用于新生儿和婴儿口服免疫的甲型副伤寒疫苗载体。我们将使用这个S。副伤寒除了一个沙门氏菌外，伤寒载体构建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.