Recombinant Attenuated Salmonella Vaccines for Humans

项目摘要/摘要 在过去十年平均每年5700万人的死亡中，约有1480万人直接死于 数以百万计的传染病是由感染的继发性影响造成的()。一年的数量 因感染伤寒沙门氏菌和甲型副伤寒沙门氏菌、乙肝病毒、链球菌而死亡 肺炎和结核分枝杆菌(Mtb)差异很大，但可能占约40% 总死亡人数，代价更大的原因是与这些疾病相关的严重发病率(241)。 认为改善健康、营养和经济福祉(后者取决于前两者) 提供提高全球生活质量的最佳手段，从而减少导致 针对战争和恐怖行为，我们提出了一个基于我们最新技术的疫苗开发计划 重组减毒沙门氏菌疫苗的研究进展我们的具体目标是：(一) 增强对RASV表达的保护性肺炎球菌的黏膜和系统抗体反应的诱导 抗原(作为模型抗原)通过增加膜泡的产生和释放以及通过改变 RASV细胞黏附属性，(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和/或完全溶解， (Iii)设计和构建甲型副伤寒沙门氏菌作为抗原和DNA疫苗输送载体 用于婴儿和成人免疫时确保安全性和有效性的基因指定特征 预防肠热病和由乙肝病毒引起的其他传染病，(Iv)设计、 甲型副伤寒沙门氏菌重组延迟裂解疫苗的构建与评价 在免疫个体的效应器淋巴组织内传递乙肝病毒(HBV)核心颗粒 融合前S_1、前S_2和T细胞表位及编码S小分子抗原的核酸疫苗在猪体细胞中的合成 免疫宿主，其特征是能够在出生后不久分娩，以及(V)设计、构建和评估 重组减毒伤寒沙门氏菌疫苗预防结核分枝杆菌感染的研究 II型和III型分泌和/或通过体内调节的延迟裂解，裂解发生在细胞室中 在体内通过MHC I类或II类和调节的延迟裂解增强抗原提呈 细胞质隔间释放针对核靶向优化的DNA疫苗，导致合成和 免疫个体对保护性抗原的修饰。我们还将开发我们的主文件，准备 并充分描述候选疫苗主种的稳定性和安全性，准备和提交方案 IRB批准，提交获得IND所需的信息，并执行安排所需的任何其他工作 最好的候选疫苗在人类志愿者身上进行临床评估。项目叙事 提出的研究将进一步发展和完善生活的设计和施工技术 重组减毒沙门氏菌疫苗(RASV)载体无针口服接种后的接种 保护性抗原和/或DNA疫苗，以刺激黏膜、系统和细胞免疫 细菌、病毒和寄生虫感染性疾病制剂。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口服携带多种乙肝病毒抗原的廉价疫苗 为新生儿和婴儿接种疫苗，以防止乙肝病毒在分娩期间传播。我们还将开发一种 RASV以伤寒沙门氏菌为载体，预防结核分枝杆菌的传播和感染。

Project Summary/Abstract 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 (64). 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 40 percent of the total deaths with an even greater cost being due to the severe morbidity associated with these diseases (241). 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 an additional infectious disease such as caused by HBV, (iv) to design, construct and evaluate a recombinant attenuated S. Paratyphi A vaccine exhibiting regulated delayed lysis within effector lymphoid tissues in the immunized individual to deliver hepatitis B virus (HBV) core particles with fused preS1, preS2 and T-cell epitopes and a DNA vaccine encoding small S antigen for synthesis in cells of the immunized host with features to enable delivery soon after birth, and (v) to design, construct and evaluate a recombinant attenuated S. Typhi vaccine 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 develop our Master File, 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. Project Narrative 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 inexpensive vaccine using S. Paratyphi A to deliver multiple hepatitis B virus (HBV) antigens for oral immunization of newborns and infants to prevent HBV transmission during childbirth. We will also develop a RASV using S. Typhi as vector to prevent transmission and infection by Mycobacterium tuberculosis.