Role of molecular drivers in memory group 1 CD1-restricted T cell differentiation and Mycobacterium tuberculosis infection

分子驱动因素在记忆组 1 CD1 限制性 T 细胞分化和结核分枝杆菌感染中的作用

• 批准号: 10491664
• 负责人: Eva Morgun
• 金额: $ 4.5万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-15 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10491664
• 关键词: Adoptive Transfer; Adult; Affect; Animal Model; Antibiotics; Antibody Formation; Antigen-Presenting Cells; Antigens; Attenuated Vaccines; B cell differentiation; B-Lymphocytes; BCG Live; BCL1 Oncogene; BCL6 gene; Bacteria; Behavior; CD1 Antigens; CRISPR/Cas technology; Cause of Death; Cell Wall; Cell physiology; Cells; Data; Development; Engineering; Epidemic; Exhibits; Family; Gene Expression Profile; Genetic Polymorphism; HIV Seropositivity; Helper-Inducer T-Lymphocyte; Human; Immune response; Immunity; Immunize; Immunocompromised Host; Immunologic Memory; Individual; Infection; Lipids; MHC Class I Genes; Memory; Molecular; Multi-Drug Resistance; Multiple drug resistant Mycobacteria Tuberculosis; Mus; Mycobacterium bovis; Mycobacterium tuberculosis; Mycolic Acid; Patients; Pattern; Peptides; Phase; Phenotype; Play; Population; Production; Proteins; Public Health; Reproducibility; Research; Role; Schools; Structure of germinal center of lymph node; Subunit Vaccines; Surface; T cell differentiation; T memory cell; T-Cell Activation; T-Cell Receptor; T-Lymphocyte; Testing; Transgenic Mice; Transgenic Organisms; Tuberculosis; Tuberculosis Vaccines; Vaccinated; Vaccination; Vaccines; Virulent; Work; anti-PD-1; anti-PD1 antibodies; antigen-specific T cells; base; cell behavior; cytokine; experience; experimental study; genetic signature; in vivo; knock-down; mouse model; mycobacterial; nanoparticle; prevent; programmed cell death protein 1; receptor; resistant strain; response; single cell sequencing; transcription factor; transcriptome; transcriptome sequencing; vaccine candidate; vaccine strategy

Project Summary Tuberculosis remains a serious global epidemic and with the rise of multi-drug resistant strains, an efficacious vaccine solution is imperative. Synthetic subunit vaccines, which do not contain live bacteria, present numerous advantages over attenuated vaccines by offering a safer option for immunocompromised individuals and being amenable to rapid, scalable, and reproducible production. The subunit vaccines currently being developed for Mycobacterium tuberculosis (Mtb) use peptide or protein antigens, which target MHC-restricted conventional T cells, overlooking the potential of Mtb lipid antigens as vaccine candidates. Mycobacterial lipids are presented by group 1 CD1 (CD1a, b, c) and group 2 CD1d to cognate T cells. Group 1 CD1-restricted T cells can be identified in patients with TB and have been shown to contribute to protective immunity against Mtb infection. In order to study further study group 1 CD1-restricted Mtb response, we previously created CD1 expressing transgenic mice (hCD1Tg) and CD1b-restricted T cell receptor transgenic T cells (DN1Tg) specific for the dominant Mtb lipid antigen mycolic acid (MA). We have since showed that DN1 T cells can be activated in vivo by vaccination with nanoparticles (NP) loaded with MA. We immunized hCD1Tg-DN1 mice with MA-NP to generate memory DN1 T cells. Using transcriptome analysis, we then determined that memory DN1 T cells most closely resemble T follicular helper (TFH) cells. In this proposal, we will assess whether two key TFH cell molecules, Bcl6 and PD-1, are necessary for memory DN1 T cell differentiation. We will also validate findings in the polyclonal setting using single cell sequencing. Finally, we will test whether memory DN1 T cell can provide protection in Mtb infection conditions. Through this work, we will be able to understand unique determinants of memory group 1 CD1-restricted T cells differentiation and their role in Mtb infection bringing us a step closer to creating a lipid-based vaccine for Mtb.

项目概要 结核病仍然是一种严重的全球流行病，并且随着多重耐药菌株的增加，有效的治疗方法 疫苗解决方案势在必行。不含活细菌的合成亚单位疫苗 与减毒疫苗相比具有许多优势，为免疫功能低下的个体提供更安全的选择 并能够进行快速、可扩展和可重复的生产。目前正在研发的亚单位疫苗 专为结核分枝杆菌 (Mtb) 开发，使用肽或蛋白质抗原，针对 MHC 限制 传统 T 细胞，忽视了 Mtb 脂质抗原作为候选疫苗的潜力。分枝杆菌脂质 由第 1 组 CD1（CD1a、b、c）和第 2 组 CD1d 呈递给同源 T 细胞。第 1 组 CD1 限制性 T 可以在结核病患者体内鉴定出细胞，并且已证明细胞有助于针对结核病的保护性免疫 结核分枝杆菌感染。为了进一步研究研究组 1 CD1 限制的 Mtb 反应，我们之前创建了 CD1 表达转基因小鼠（hCD1Tg）和CD1b限制性T细胞受体转基因T细胞（DN1Tg）特异性 为主要的 Mtb 脂质抗原分枝菌酸 (MA)。此后我们证明 DN1 T 细胞可以被激活 通过接种负载 MA 的纳米颗粒 (NP) 进行体内接种。我们用 MA-NP 免疫 hCD1Tg-DN1 小鼠 产生记忆 DN1 T 细胞。通过转录组分析，我们确定记忆 DN1 T 细胞 与滤泡辅助 T (TFH) 细胞最为相似。在本提案中，我们将评估两个关键的 TFH 细胞是否 Bcl6 和 PD-1 分子是记忆 DN1 T 细胞分化所必需的。我们还将验证调查结果 在多克隆环境中使用单细胞测序。最后，我们将测试记忆DN1 T细胞是否可以 在 Mtb 感染条件下提供保护。通过这项工作，我们将能够了解独特的 记忆组 1 CD1 限制性 T 细胞分化的决定因素及其在 Mtb 感染中的作用 距离研制结核分枝杆菌脂质疫苗又近了一步。