Harnessing B cells for TB vaccine development to improve therapy of TB and TB-HIV coinfection

利用 B 细胞开发结核病疫苗，以改善结核病和结核病-艾滋病毒合并感染的治疗

• 批准号: 10662211
• 负责人: Martin Alfons Gengenbacher
• 金额: $ 127.65万
• 依托单位: HACKENSACK UNIVERSITY MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10662211
• 关键词: Adult; Animal Model; Animals; Area; Attenuated Vaccines; B-Cell Activation; B-Cell Development; B-Lymphocyte Subsets; B-Lymphocytes; BCG Live; BCG Vaccine; Blood; Cell secretion; Cells; Child; Childhood; Circulation; Clinical; Combination Drug Therapy; Complement; Containment; Data; Development; Drug resistance; Exposure to; Face; Flow Cytometry; Frequencies; Genes; Genetic; Genetic Engineering; HIV; HIV therapy; HIV/TB; Human; Immunodeficient Mouse; Immunologics; Immunophenotyping; Individual; Infection; Infection prevention; Inhibition of Apoptosis; Ligands; Longevity; Lung; Macrophage; Measures; Memory; Modeling; Monitor; Mus; Mycobacterium bovis; Mycobacterium tuberculosis; Organ; Patients; Peripheral Blood Mononuclear Cell; Persons; Pharmaceutical Preparations; Pharmacotherapy; Phenotype; Pre-Clinical Model; Proliferating; Recombinants; Recurrence; Regimen; Relapse; Research; Role; Route; Safety; T cell response; T-Lymphocyte; Testing; Therapeutic; Time; Treatment Efficacy; Tuberculosis; Tuberculosis Vaccines; Vaccinated; Vaccination; Vaccine Research; Vaccines; Virus Diseases; adaptive immunity; antiretroviral therapy; chemotherapy; co-infection; cohort; cytokine; experimental study; genetic resistance; humoral immunity deficiency; immunogenic; immunogenicity; immunoregulation; improved; infected B cell; inhibition of autophagy; innovation; knock-down; mouse model; mutant; mycobacterial; next generation; novel; pathogen; prevent; prophylactic; protective efficacy; rBCG; reactivation from latency; receptor; relapse prevention; response; safety assessment; therapeutic vaccine; tuberculosis drugs; tuberculosis treatment; vaccine candidate; vaccine development; vaccine evaluation

Abstract Current tuberculosis (TB) control in the presence or absence of human immunodeficiency virus (HIV) infection is suboptimal: Bacillus Calmette-Guérin (BCG), the sole TB vaccine in clinical use, provides only limited protection during childhood and multi-drug chemotherapy faces challenges due to the alarming spread of drug resistance. An integrated approach combining chemotherapy with a therapeutic vaccine given after exposure to Mycobacterium tuberculosis (Mtb) could be a way forward. TB vaccine research mostly focused on prophylactic candidates aiming to prevent infection. Therapeutic TB vaccines with novel mechanisms of action are needed (i) to complement existing chemotherapy and prevent relapse thereafter, and (ii) to cure latent TB infection (LTBI) or prevent reactivation. T lymphocytes are critical for the control of TB infection and the specific T cell response elicited by a vaccine has been used as a key correlate of immunogenicity. In contrast, B cells are understudied in all areas of TB research. Using a deep immunophenotyping approach, our preliminary data in mice suggest that TB infection leads to dramatic changes in the landscape of B cell subpopulations. We identified a novel B cell subset with marginal zone (MZ) phenotype that was activated, had a memory phenotype and expressed receptors recognizing the human cytokines A Proliferation-Inducing Ligand, APRIL, and B cell Activating Factor, BAFF, critical for B cell development and survival. Functional studies indicated that murine MZ B cells contributed to Mtb containment in mice. Surprisingly, these B cells expressed a panel of Th1 cytokines, well studied in T cells, suggesting a possible role in the first line of defense against TB infection. We found that MZ B cells were depleted in blood of TB patients and TB/HIV coinfected people. Our hypothesis is that MZ B cells can be restored by antitubercular therapy and harnessed for TB vaccine development. To test this, we genetically engineered BCG to express the human cytokines APRIL and BAFF that stimulate development and longevity of BAFF receptor- and APRIL receptor-expressing B cells, including MZ B cells. We will determine the safety, immunogenicity and prophylactic efficacy in mice. The therapeutic efficacy will be evaluated in two novel innovative mouse models that allow us to determine the capacity of cytokine expression in BCG strains to prevent relapse after drug treatment and to reduce the reactivation frequency of paucibacillary TB mimicking aspect of LTBI in humans. We will determine the frequency of MZ B cells in peripheral blood mononuclear cells (PBMC) from people having LTBI and TB, across HIV status, including antitubercular and antiretroviral therapy. We will stimulate PBMC of the same cohorts with cytokine-expressing BCG strains to study the immunological consequences and to demonstrate, in principle, the feasibility of these vaccines during therapy of TB, HIV and TB/HIV coinfection. We will monitor the global landscapes of T cells and B cells by high parameter flow cytometry to define immunological correlates of TB infection and vaccine protection. Our proposal will determine whether B cells can be harnessed by cytokine-secreting B cell-targeting BCG vaccines for TB vaccine development.

摘要 在存在或不存在人类免疫缺陷病毒(HIV)感染的情况下目前的结核病控制 是次优的：卡介苗(BCG)，临床上唯一使用的结核病疫苗，只能提供有限的 由于毒品的惊人传播，儿童时期的保护和多种药物化疗面临挑战 抵抗。一种将化疗与暴露于禽流感后给予治疗性疫苗相结合的综合方法 结核分枝杆菌(Mtb)可能是一个前进的方向。结核病疫苗研究主要集中在预防方面 候选人的目标是防止感染。需要具有新作用机制的治疗性结核病疫苗 (I)补充现有的化疗，防止化疗后复发；及。(Ii)治愈潜伏的结核病感染。 或阻止重新激活。T淋巴细胞对控制结核病感染和特异性T细胞反应至关重要 疫苗诱导的免疫原性已被用作免疫原性的关键相关因素。相比之下，B细胞研究不足 在结核病研究的所有领域。使用深度免疫表型方法，我们在小鼠身上的初步数据表明 结核病感染导致了B细胞亚群格局的戏剧性变化。我们发现了一部小说B 具有边缘带(MZ)表型的细胞亚群被激活，具有记忆表型并表达 识别人类细胞因子A、增殖诱导配体、APRIL和B细胞激活因子的受体 Baff，对B细胞的发育和生存至关重要。功能研究表明，小鼠的MZ B细胞对 在小鼠体内控制结核分枝杆菌。令人惊讶的是，这些B细胞表达一组Th1细胞因子，在T细胞中得到了很好的研究 细胞，这表明它可能在预防结核病感染的第一道防线中发挥作用。我们发现MZ B细胞是 结核病患者和结核病/艾滋病毒混合感染者的血液耗尽。我们的假设是MZ B细胞可以被修复 通过抗结核治疗，并用于结核病疫苗的开发。为了测试这一点，我们进行了基因工程 卡介苗表达促进BAFF发育和长寿的人细胞因子APRIL和BAFF 表达受体和APRIL受体的B细胞，包括MZ B细胞。我们将确定其安全性， 小鼠的免疫原性和预防效果。治疗效果将在两部小说中进行评估 创新的小鼠模型，使我们能够确定卡介苗菌株中细胞因子的表达能力，以防止 在药物治疗后复发和降低少杆菌结核再激活频率方面类似于 人类的LTBI。我们将测定外周血单个核细胞(PBMC)中MZ B细胞的频率 来自患有LTBI和TB的人，包括抗结核和抗逆转录病毒治疗。我们会 用表达细胞因子的卡介苗刺激同一队列的PBMC进行免疫学研究 并在原则上证明这些疫苗在结核病、艾滋病毒和艾滋病治疗期间的可行性。 结核病/艾滋病毒混合感染。我们将用高参数流式细胞术监测T细胞和B细胞的全球景观 明确结核病感染与疫苗保护的免疫学相关性。我们的提案将决定是否 B细胞可以被分泌细胞因子的B细胞靶向卡介苗用于结核病疫苗的开发。