The Role of Canonical and Non-canonical Autophagy in B Cell Immunity

典型和非典型自噬在 B 细胞免疫中的作用

• 批准号: 10349500
• 负责人: Facundo Damian Batista
• 金额: $ 45.56万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-03-01 至 2024-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10349500
• 关键词: Ablation; Affect; Affinity; Antibodies; Antibody Affinity; Antigen Presentation; Antigens; Autoimmune Diseases; Autophagocytosis; B Cell Proliferation; B cell differentiation; B-Cell Activation; B-Lymphocytes; Biology; Cell Survival; Cell physiology; Cells; Cellular biology; Complex; Data; Degradation Pathway; Development; Down-Regulation; Equilibrium; Gene Deletion; Generations; Genes; Genetic; Goals; HIV-1; Health; Homeostasis; Human; Humoral Immunities; Immune; Immune Tolerance; Immune response; Immune system; Immunity; Immunization; Immunoglobulin Class Switching; Impairment; Infection; Infectious Agent; Influenza; Knowledge; Lead; Light; Link; Longevity; Maintenance; Memory; Memory B-Lymphocyte; Metabolic; Metabolism; Mitochondria; Molecular; Mus; Outcome Study; Pathway interactions; Phosphotransferases; Physiological; Plasma Cells; Play; Process; Production; Public Health; Regulation; Role; Shapes; Signal Transduction; Structure of germinal center of lymph node; T-Lymphocyte; Testing; Therapeutic Monoclonal Antibodies; Time; Vaccination; Vaccines; antigen binding; experimental study; fighting; immune function; immunosenescence; improved; in vivo; insight; leukemia; novel; novel therapeutics; novel vaccines; pathogen; protein degradation; response; trafficking; vaccination strategy

B cells play a central role in human health by protecting us against infections through their ability to produce high-affinity antibodies and long-lived memory cells capable of antigenic recall. Perturbations in B cell function can not only lead to lowered ability to fight infections and inefficacy of vaccinations, but also to conditions such as leukemia, and autoimmune diseases. Thus, understanding the molecular mechanisms that contribute to optimal B cell function is extremely important for the generation of new therapeutics and vaccination strategies. The major goal of this proposal is to study how autophagy, a lysosomal degradative pathway with numerous physiological and pathophysiological roles, shapes B cell immunoresponses. The induction of canonical autophagy typically requires the downregulation of the biosynthetic kinase, mTORC1. However, in B cells upon BCR stimulation both autophagy and mTORC1 activity are simultaneously upregulated, suggesting that B cell autophagy is likely to be mTORC1 independent. Our lab has recently shown that there is a switch from basal canonical autophagy in naïve, antigen-inexperienced B cells to an unconventional, mTORC1-independent, non- canonical activity in the germinal center (GC) B cells upon antigenic stimulation. GCs are the determinants of high affinity, class switched antibody generation, and therefore the study of the mechanisms that drive these high-quality immunoresponses is vitally important in the development of next-generation vaccines, therapeutic monoclonal antibodies and in the treatment of autoimmune diseases. Our working hypothesis is that non-canonical autophagy is important during B cell proliferation and will be relevant to GC biology affecting antibody quality, while canonical autophagy is important for long-lived memory cells and plasma cells influencing antibody durability. To test this hypothesis, we will evaluate GC and long-term immune responses in mice bearing genetic ablations of novel autophagy genes that we know affect the balance between canonical and non-canonical autophagy, namely Wipi1, Wipi2 and Rubicon. We will also determine the mechanisms by which the deletions of these genes affect antigen presentation and in eliciting T cell help. Finally, because autophagy intersects with mitochondrial integrity and metabolism, we will determine how these gene deletions control the capacity of GC B cells, memory and plasma cells to regulate ROS production, mitochondrial homeostasis and metabolic status and potentially influence B cell fate. We envision that the successful completion of these experimental aims will not only provide a better understanding of B cell function, but also mechanistic insights into how to modulate autophagy to affect humoral immunity – augment it in cases such as immunosenescence, or downregulate it in instances of autoimmune diseases.

B细胞在人类健康中发挥着核心作用，通过它们的生产能力保护我们免受感染。 高亲和力抗体和能够回忆抗原的长寿记忆细胞。B细胞功能的扰动 不仅会导致抵抗感染的能力降低和疫苗接种无效， 如白血病和自身免疫性疾病。因此，了解有助于 最佳的B细胞功能对于产生新的治疗剂和疫苗接种策略是极其重要的。 这项提议的主要目标是研究自噬，一种具有许多细胞因子的溶酶体降解途径， 生理和病理生理作用，形成B细胞免疫应答。典范归纳法 自噬通常需要下调生物合成激酶mTORC 1。然而，在B细胞中， BCR刺激自噬和mTORC 1活性同时上调，表明B细胞 自噬可能是mTORC 1独立的。我们的实验室最近发现， 在幼稚的、无抗原经验的B细胞中，典型的自噬作用转变为非常规的、mTORC 1非依赖性的、非特异性的自噬作用。 在抗原刺激后，在生发中心（GC）B细胞中的典型活性。GC是决定 高亲和力，类别转换抗体的产生，因此研究驱动这些的机制， 高质量的免疫应答对于开发下一代疫苗、治疗性疫苗和免疫治疗性疫苗至关重要。 单克隆抗体和治疗自身免疫性疾病。 我们的工作假设是，非典型的自噬在B细胞增殖过程中很重要， 与影响抗体质量的GC生物学相关，而典型的自噬对于长期记忆很重要 影响抗体持久性的细胞和浆细胞。为了验证这一假设，我们将评估GC和长期 我们知道，携带新型自噬基因的小鼠的免疫反应会影响平衡， 经典和非经典自噬之间的差异，即Wipi 1，Wipi 2和Rubicon。我们还将确定 这些基因的缺失影响抗原呈递和引发T细胞帮助的机制。最后， 由于自噬与线粒体的完整性和代谢交叉，我们将确定这些基因是如何表达的。 缺失控制GC B细胞、记忆细胞和浆细胞调节ROS产生、线粒体 稳态和代谢状态并潜在地影响B细胞命运。 我们设想，这些实验目标的成功完成不仅将提供一个更好的 了解B细胞的功能，而且还对如何调节自噬以影响体液免疫的机制有深入的了解。 免疫-在免疫衰老情况下增强它，或在自身免疫性疾病的情况下下调它 疾病

项目成果

Dynamic reorganisation of intermediate filaments coordinates early B-cell activation.

中间丝的动态重组协调早期 B 细胞激活。

• DOI: 10.26508/lsa.201800060
• 发表时间: 2018
• 期刊: Life science alliance
• 影响因子: 4.4
• 作者: Tsui,Carlson;Maldonado,Paula;Montaner,Beatriz;Borroto,Aldo;Alarcon,Balbino;Bruckbauer,Andreas;Martinez-Martin,Nuria;Batista,FacundoD
• 通讯作者: Batista,FacundoD

One-step CRISPR/Cas9 method for the rapid generation of human antibody heavy chain knock-in mice.

• DOI: 10.15252/embj.201899243
• 发表时间: 2018-09-14
• 期刊: The EMBO journal
• 作者: Lin YC;Pecetta S;Steichen JM;Kratochvil S;Melzi E;Arnold J;Dougan SK;Wu L;Kirsch KH;Nair U;Schief WR;Batista FD
• 通讯作者: Batista FD

Protein Kinase C-β Dictates B Cell Fate by Regulating Mitochondrial Remodeling, Metabolic Reprogramming, and Heme Biosynthesis.

• DOI: 10.1016/j.immuni.2018.04.031
• 发表时间: 2018-06-19
• 期刊: Immunity
• 影响因子: 32.4
• 作者: Tsui C;Martinez-Martin N;Gaya M;Maldonado P;Llorian M;Legrave NM;Rossi M;MacRae JI;Cameron AJ;Parker PJ;Leitges M;Bruckbauer A;Batista FD
• 通讯作者: Batista FD