Cytokine control of Red Blood Cell Alloimmunization

红细胞同种免疫的细胞因子控制

• 批准号: 8424288
• 负责人: CHANCE MARION JOHN LUCKEY
• 金额: $ 19.71万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-02-15 至 2013-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8424288
• 关键词: Address; Alleles; Alloimmunization; Antibody Formation; Antigens; Automobile Driving; B cell differentiation; B-Cell Development; B-Lymphocytes; Biochemical Genetics; Blood; CD4 Positive T Lymphocytes; Cell Differentiation process; Cells; Cessation of life; Chromatin; Chromatin Remodeling Factor; Clinical; Complex; Cytokine Signaling; Development; Diagnostic; Erythrocyte Transfusion; Erythrocytes; Event; Generations; Genetic Programming; Helper-Inducer T-Lymphocyte; Human; Immunization; Immunoglobulin-Secreting Cells; Infectious Agent; Interleukin-6; Isoantibodies; Lead; Life; Lymphocyte; Maintenance; Measures; Medicine; Modeling; Molecular; Molecular Target; Morbidity - disease rate; Mouse Strains; Mus; PI3K/AKT; Patients; Phosphorylation; Post-Translational Protein Processing; Process; Production; Reaction; Research; Resources; STAT3 gene; Series; Signal Pathway; Signal Transduction; Staging; Sterility; T cell differentiation; T-Cell Activation; T-Lymphocyte; Testing; Therapeutic; Time; Transfusion; Vaccination; Work; cellular targeting; clinically relevant; cytokine; human FRAP1 protein; in vivo; mTOR Signaling Pathway; member; mortality; mouse model; novel; receptor; research study; response; tool; transcription factor

DESCRIPTION (provided by applicant): RBC alloimmunization remains a common clinical problem in transfusion medicine. Alloantibodies are responsible for both immediate and delayed hemolytic transfusion reactions; leading to substantial morbidity and occasional mortality. For those patients unfortunate enough to make multiple alloantibodies, provision of compatible RBCs can be both time and resource intensive. In some cases, this can result in an inability to locate an otherwise life-saving therapy. Despite its clinical importance, the fundamental molecular signals that drive RBC immunization remain unclear. Like infectious agents, the generation of alloantibodies to RBCs requires the activation of antigen specific CD4+ T cells and B cells, where naove CD4+ T cells differentiate into T Follicular Helper cells (TFH) that are essential for complete B cell differentiation into antibody secreting cells. However, our understanding of the molecular mechanisms by which naove CD4+ T cells are induced to differentiate and express a TFH genetic program "in vivo" remains partial. Furthermore, the molecular factors responsible for naove T cell activation and TFH differentiation by the relatively weak stimulant of RBC transfusion are unknown. If we are to ever successfully intervene in this pathogenic process, we must first identify the cellular targets and molecular signals responsible for RBC alloantibody generation. The development of clinically relevant mouse models of RBC storage and transfusion provides a novel set of tools to address this issue. Recent work in these models demonstrated that RBC alloimmunization is enhanced by RBC storage, and that transfusion of stored RBCs significantly increases the systemic production of multiple cytokines including IL-6. We have subsequently demonstrated that i) IL-6 deficient mice are less susceptible to RBC alloimmunization in both fresh and stored blood settings, ii) IL-6 signaling activates the PI3K/AKT/mTOR signaling pathway in naove CD4+ T cells, and iii) induction of PI3K/AKT/mTOR signals correlates with increased phosphorylation of multiple members of the BAF chromatin remodeling complex in primary naove T cells. We therefore propose to identify the specific cellular targets of IL-6 in RBC alloimmunization, and further determine the molecular signals in naove CD4+ T cells induced by IL-6. The objective of our research plan is to test the specific hypothesis that the IL-6 induced by transfusion of fresh and stored RBCs drives TFH differentiation from naove CD4+ T cells. Furthermore, we hypothesize that IL-6 signaling induces phospho-modulation of the BAF chromatin remodeling complex via the action of the PI3K/AKT/mTOR signaling pathway. We will test our hypothesis by combining a well established mouse model of RBC alloimmunization and storage with recently developed mouse strains expressing a floxed allele of IL-6 Receptor (IL-6RA). In the fullness of time, we anticipate that identification of the molecular circuitry driving RBC antibody production will provide therapeutic and diagnostic targets for further clinical development.

描述（由申请人提供）：RBC同种免疫仍然是输血医学中常见的临床问题。同种抗体负责直接和延迟的溶血输血反应。导致大量发病率和偶尔死亡率。对于那些不幸的患者来说，足以制造多种同种抗体，提供兼容的RBC可能是时间和资源密集型。在某些情况下，这可能导致无法定位原本挽救生命的疗法。尽管具有临床意义，但驱动RBC免疫的基本分子信号仍不清楚。像传染剂一样，RBC的同种抗体的产生需要激活抗原特异性CD4+ T细胞和B细胞，其中NaOVE CD4+ T细胞分化为T卵泡辅助细胞（TFH），这对于完全B细胞分化为抗体分泌细胞至关重要。但是，我们对NAOVE CD4+ T细胞的分子机制的理解被诱导分化并表达TFH遗传程序“体内”仍然部分。此外，相对较弱的RBC输血刺激剂，导致NAOVE T细胞激活和TFH分化的分子因子尚不清楚。如果我们要成功干预这一病原过程，则必须首先确定导致RBC同种抗体产生的细胞靶标和分子信号。 RBC存储和输血的临床相关小鼠模型的开发提供了一组新的工具来解决此问题。这些模型中的最新工作表明，RBC同种免疫通过RBC储存增强，并且储存的RBC的输血显着增加了包括IL-6在内的多种细胞因子的全身产生。随后，我们证明了I）IL-6缺乏小鼠在新鲜和储存的血液环境中不太容易受到RBC同种异体免疫性的影响，II）IL-6信号传导在Naove CD4+ T细胞中NAOVE CD4+ T细胞中NAOVE的pi3k/akt/mTOR信号通路，以及iii的多种构建范围，以及III的构建范围。原代NAOVE T细胞中的染色质重塑复合物。因此，我们建议在RBC同种免疫中确定IL-6的特定细胞靶标，并进一步确定IL-6诱导的NaOVE CD4+ T细胞中的分子信号。我们的研究计划的目的是检验以下特定假设：由新鲜和存储的RBC输血诱导的IL-6驱动Naove CD4+ T细胞的TFH分化。此外，我们假设IL-6信号传导通过PI3K/AKT/MTOR信号通路的作用诱导BAF染色质重塑复合物的磷酸化调节。我们将通过结合良好的RBC同种免疫化小鼠模型和最近开发的小鼠菌株来检验我们的假设，该模型表达了IL-6受体（IL-6RA）的flox等位基因的小鼠菌株。在时间的饱满状态下，我们预计鉴定驱动RBC抗体产生的分子电路将为进一步的临床发育提供治疗和诊断靶标。