A novel hyper-immunogenic low virulent BCG vaccine against tuberculosis

一种新型高免疫原性低毒力结核病卡介苗疫苗

• 批准号: 10639030
• 负责人: SUBRAMANIAN DHANDAYUTHAPANI
• 金额: $ 46.24万
• 依托单位: TEXAS TECH UNIVERSITY HEALTH SCIENCES CENTER AT EL PASO
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-02-03 至 2028-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10639030
• 关键词: Adult; Animal Model; Antibiotics; Antigen Presentation; Antigen-Presenting Cells; Apoptosis; Attenuated; Attenuated Vaccines; Autophagocytosis; BCG Live; BCG Vaccine; C57BL/6 Mouse; Cavia; Cell Culture Techniques; Cells; Cessation of life; Child; Childhood; DNA; Dendritic Cells; Disease; Dose; Ensure; Failure; Gene Deletion; Generations; Genes; Goals; HIV; HIV Infections; HIV-1; Health; Immune; Immune response; Immunity; Immunize; Immunocompetent; Immunologic Deficiency Syndromes; In Vitro; Individual; Infant; Knock-out; Licensing; Lysosomes; Macrophage; Meta-Analysis; Methods; Mus; Mycobacterium bovis; Mycobacterium tuberculosis; Outcome; Persons; Phagosomes; Phase III Clinical Trials; Plasmids; Preventive; Process; Protein Subunits; Proteins; Pulmonary Tuberculosis; Recombinant Vaccines; Recombinants; SCID Mice; Safety; Spleen; Structure of parenchyma of lung; T memory cell; Testing; Time; Tissues; Tuberculosis; Tuberculosis Vaccines; Vaccines; Viral Vector; Virulence; Virulent; antigen processing; cell mediated immune response; efficacy evaluation; genetic manipulation; genome integrity; genome sequencing; global health; homologous recombination; human morbidity; human mortality; humanized mouse; immune RNA; immunogenic; immunogenicity; improved; in vivo; infant infection; innovation; knockout gene; mouse model; mutant; novel; overexpression; pathogenic bacteria; prevent; safety assessment; study population; suicidal; transcriptome sequencing; transcriptomics; vaccine access; vaccine candidate; vaccine trial; whole genome

SUMMARY Human morbidity and mortality due to tuberculosis (TB), caused by the bacterial pathogen Mycobacterium tuberculosis (Mtb), continue to be of significant health concern throughout the world. Bacille Calmette-Guerin (BCG) still remains the only approved vaccine against TB. Although BCG is considered safe and partially effective against extra-pulmonary childhood TB, its ability to protect against childhood and adult pulmonary TB is still questionable. In addition, there is a concern that BCG does not induce long lasting immune responses in the immunized individuals. However, our findings with Mtb- and BCG- derived recombinant vaccines strongly suggest that BCG can be improved to be more efficacious against TB. The goal of this proposal is to improve BCG by rationally deleting genes. We plan to sequentially delete three genes of BCG namely sapM, zmp1 and nuoG, through homologous recombination, to result in a triple knockout BCG (BCG-TKO) strain. These genes encode proteins that enable BCG to evade host immune response by preventing phagosome-lysosomal fusion, autophagy, apoptosis and other related processes in the antigen presenting cells (APCs) such as dendritic cells and macrophages. We hypothesize that deletion of these genes will allow the BCG-TKO strain to be efficiently processed by APCs, which will lead to increased antigen presentation to the immune cells and enhanced in vivo immunogenicity and efficacy. Additionally, we anticipate that deletion of these genes will reduce the virulence of the BCG, thus making the BCG-TKO to be HIV safe. We plan to accomplish our goal with three aims: 1) construct a BCG-TKO strain through homologous recombination, 2) analyze the immunogenicity and safety of the BCG-TKO strain in a SCID mouse model, and 3) investigate the efficacy of the BCG-TKO in the regular and humanized mouse models with and without HIV infection. Overall, we expect that this proposal will produce a highly efficacious, third generation BCG vaccine against TB that will be suitable for administration even in HIV infected infants.

概括 由细菌病原体分枝杆菌引起的结核病 (TB) 导致的人类发病率和死亡率 结核病（Mtb）仍然是全世界严重的健康问题。卡介苗 （卡介苗）仍然是唯一获得批准的结核病疫苗。尽管卡介苗被认为是安全的并且部分 有效对抗儿童肺外结核病，具有预防儿童和成人肺结核的能力 仍存疑问。此外，还有人担心卡介苗不会在人体中诱导持久的免疫反应。 已免疫的个体。然而，我们对 Mtb 和 BCG 衍生的重组疫苗的研究结果强烈 表明可以改进卡介苗以更有效地对抗结核病。 该提案的目标是通过合理删除基因来改进BCG。我们计划依次删除 BCG的三个基因，即sapM、zmp1和nuoG，通过同源重组，产生三重基因 敲除卡介苗（BCG-TKO）菌株。这些基因编码的蛋白质使卡介苗能够逃避宿主免疫 通过阻止吞噬体-溶酶体融合、自噬、细胞凋亡和其他相关过程来响应 抗原呈递细胞（APC），例如树突状细胞和巨噬细胞。我们假设删除 这些基因将使 BCG-TKO 菌株能够被 APC 有效加工，从而导致增加 抗原呈递给免疫细胞并增强体内免疫原性和功效。此外，我们 预计删除这些基因将降低 BCG 的毒力，从而使 BCG-TKO 成为可能。 艾滋病毒安全。我们计划通过三个目标来实现我们的目标：1）通过同源构建BCG-TKO菌株 重组，2) 分析 BCG-TKO 株在 SCID 小鼠模型中的免疫原性和安全性，以及 3) 研究 BCG-TKO 在携带和不携带 HIV 的常规和人源化小鼠模型中的功效 感染。总体而言，我们预计该提案将生产出高效的第三代卡介苗疫苗 抗结核病，甚至适用于感染艾滋病毒的婴儿。