Generation and Regulation of Anti-Viral CD4 T Cells with Cytolytic Potential

具有溶细胞潜力的抗病毒 CD4 T 细胞的生成和调节

• 批准号: 8705820
• 负责人: Deborah M. Brown
• 金额: $ 35.13万
• 依托单位: UNIVERSITY OF NEBRASKA LINCOLN
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-08-01 至 2015-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8705820
• 关键词: Affinity; Antibodies; Antibody Affinity; Antibody Formation; Antiviral Response; Appearance; Automobile Driving; Avian Influenza A Virus; Binding; CD4 Positive T Lymphocytes; CD8B1 gene; Capsid Proteins; Cells; Cellular Immunity; Chronic; Communicable Diseases; Cytokine Receptors; Cytokine Signaling; Cytomegalovirus; Data; Development; Emerging Communicable Diseases; Enhancing Antibodies; Epitopes; Family suidae; Generations; Goals; H7N7; HIV; Health; Hemagglutinin; Human; Human Herpesvirus 4; Immune; Immune response; Immune system; Immunity; In Vitro; Infection; Inflammatory; Influenza; Influenza A Virus, H1N1 Subtype; Influenza A Virus, H5N1 Subtype; Influenza A virus; Interleukin-2; Knockout Mice; Knowledge; Lead; Life; Lung; Lytic; Malignant Neoplasms; Mediating; Memory; Mission; Modeling; Molecular; Monitor; Mus; Mutation; Neuraminidase; Outcome; Pathway interactions; Patients; Phenotype; Phosphorylation; Physiological; Population; Proteins; Public Health; Publishing; Recovery; Regulation; Reporting; Research; Role; STAT5A gene; Signal Transduction; Site; Staging; T cell differentiation; T memory cell; T-Lymphocyte; T-Lymphocyte Subsets; Testing; Vaccine Design; Vaccines; Viral; Viral Load result; Viral Proteins; Virus; Virus Diseases; Work; base; burden of illness; combat; cytokine; cytotoxicity; design; driving force; global health; human subject; in vivo; influenzavirus; innovation; interest; lymph nodes; novel; novel vaccines; pandemic disease; pathogen; perforin; promoter; response; transcription factor

DESCRIPTION (provided by applicant): Emerging pandemic infections such as influenza A virus (IAV) and human immunodeficiency virus (HIV) remain major global health concerns due to high viral mutation rates, lack of defined immune correlates of protection and ineffective vaccines. CD4 T cells are essential in the immune response to viral infections because of their ability to enhance antibody production and help maintain CD8 T cell memory responses. Recently, a more direct role for CD4 cells during anti-viral responses has been appreciated, with many reports indicating CD4 T cells can acquire perforin mediated cytotoxicity that may be important for decreasing viral load. However, the signals that induce the differentiation of cytolytic CD4 cells (CD4 CTL) and the physiological impact of these cells in anti-viral immune responses remain obscure. The long-term goal of these studies is to understand the development of CD4 CTL and their role in vivo in order to inform vaccine strategies that target cell mediated immunity against infectious diseases. The objective of this proposal is to identify cytokine and transcription factor signals inducing CD4 CTL effector and memory responses leading to long term, protective anti- viral responses. Our central hypothesis is: class II restricted, perforin mediated cytotoxicity develops via IL-2 signaling and Th1 polarizing conditions, survives in a memory population and contributes to heterosubtypic protection. We formulated this hypothesis based on previous studies and preliminary data demonstrating that Th1 polarized CD4 effectors acquire perforin mediated cytotoxicity in response to IAV infection and a population of these cells can be identified at the memory stage. Further, IL-2 signaling via high affinity IL-2R¿ pro- motes STAT5 phosphorylation and T-bet expression, correlating with GrB and perforin mediated cytotoxicity. This hypothesis will be tested by pursuing three specific aims: 1) Identify the cytokine signals required for full CD4 CTL development, 2) Identify the transcription factors driving CD4 CTL differentiation, and 3) Establish a role for CD4 CTL in the memory pool and recall responses to influenza infection. In the first aim, the role of IL-2 and inflammatory cytokine signals in CD4 CTL development will be established using cytokine and cytokine receptor knockout mice. In aim 2, candidate transcription factors such as STAT5, T-bet and Eomes, as well as unique transcription factors identified by microarray, will be tested for their ability to bind to the perforin promoter and control CD4 CTL differentiation. Finally, aim 3, will monitor the development of CD4 CTL memory responses and their ability to promote heterosubtypic immunity by using viruses that differ in outer coat proteins in CD8 deficient mice or using viruses that lack dominant CD8 epitopes. This research is innovative because it uses a unique in vitro activation model, as well as an in vivo approach to interrogate the requirements for differentiation of a novel subset of CD4 T cells with cytolytic activity. The proposed research is significant because it is the first step in understanding the mechanisms that drive CD4 CTL differentiation that has the potential to inform novel vaccine strategies targeting T cell immunity against emerging pathogens.

描述（由申请方提供）：由于病毒突变率高、缺乏明确的免疫保护相关性和疫苗无效，甲型流感病毒（IAV）和人类免疫缺陷病毒（HIV）等新出现的大流行性感染仍然是全球主要的健康问题。CD 4 T细胞在对病毒感染的免疫应答中是必不可少的，因为它们能够增强抗体产生并帮助维持CD 8 T细胞记忆应答。最近，已经认识到CD 4细胞在抗病毒应答期间的更直接的作用，许多报告表明CD 4 T细胞可以获得穿孔素介导的细胞毒性，这对于降低病毒载量可能是重要的。然而，诱导细胞溶解性CD 4细胞（CD 4 CTL）分化的信号和这些细胞在抗病毒免疫应答中的生理影响仍然不清楚。这些研究的长期目标是了解CD 4 CTL的发展及其在体内的作用，以便为靶向细胞介导的抗感染性疾病免疫的疫苗策略提供信息。本发明的目的是鉴定诱导CD 4 CTL效应子和记忆应答的细胞因子和转录因子信号，从而导致长期的保护性抗病毒应答。我们的中心假设是：II类限制性穿孔素介导的细胞毒性通过IL-2信号传导和Th 1极化条件发展，在记忆群体中存活并有助于异亚型保护。我们制定了这一假设的基础上，以前的研究和初步数据表明，Th 1极化的CD 4效应细胞获得穿孔素介导的细胞毒性响应IAV感染和人口的这些细胞可以在记忆阶段被确定。此外，通过高亲和力IL-2 R的IL-2信号传导促进STAT 5磷酸化和T-bet表达，与GrB和穿孔素介导的细胞毒性相关。这一假设将通过追求三个具体目标来检验：1）鉴定完全CD 4 CTL发育所需的细胞因子信号，2）鉴定驱动CD 4 CTL分化的转录因子，和3）建立CD 4 CTL在记忆库和对流感感染的回忆应答中的作用。在第一个目标中，将使用细胞因子和细胞因子受体敲除小鼠建立IL-2和炎性细胞因子信号在CD 4 CTL发育中的作用。在目标2中，将测试候选转录因子如STAT 5、T-bet和Eomes以及通过微阵列鉴定的独特转录因子结合穿孔素启动子和控制CD 4 CTL分化的能力。最后，目标3， 将监测CD 4 CTL记忆应答的发展及其通过使用在CD 8缺陷小鼠中外壳蛋白不同的病毒或使用缺乏显性CD 8表位的病毒促进异亚型免疫的能力。这项研究是创新的，因为它使用了独特的体外活化模型，以及体内方法来询问具有细胞溶解活性的新型CD 4 T细胞亚群的分化要求。拟议研究 是重要的，因为它是理解驱动CD 4 CTL分化的机制的第一步，该机制有可能为靶向T细胞免疫的新型疫苗策略提供信息 对抗新出现的病原体