Innate Immune Defects in HIV-Exposed Uninfected Infants: Effect on Respiratory Syncytial Virus Infection

暴露于 HIV 的未感染婴儿的先天免疫缺陷：对呼吸道合胞病毒感染的影响

• 批准号: 10563218
• 负责人: Christiana Elizabeth Smith-Anderson
• 金额: $ 16.2万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-03-01 至 2025-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10563218
• 关键词: Address; Antigen-Presenting Cells; Bioinformatics; Blood Cells; Cell physiology; Cells; Cessation of life; Coculture Techniques; DNA Methylation; Data; Data Analyses; Defect; Disease; Endothelium; Epigenetic Process; Etiology; Exposure to; Fetus; Functional disorder; Gene Expression; Genes; Genetic Transcription; Genomics; Goals; HIV; HIV Infections; HIV-exposed uninfected infant; Health; Hospitalization; Human; Hypermethylation; Immune; Immune System Diseases; Immune system; Immunologics; Immunology; Impairment; In Vitro; Incubated; Infant; Inflammation; Inflammatory; Innate Immune Response; Interferon Type II; Interleukin-10; Interleukin-12; Interleukin-6; International; Investigation; Knowledge; Laboratories; Learning; Life; Mentors; Mission; Modification; Mononuclear; Morbidity - disease rate; Mothers; National Institute of Child Health and Human Development; Natural Killer Cells; Neonatal; Outcome; Pathway interactions; Perinatal; Peripheral Blood Mononuclear Cell; Phenotype; Plasma; Play; Population; Pregnancy; Pregnant Women; Production; Research; Research Personnel; Respiratory Syncytial Virus Infections; Respiratory Tract Infections; Respiratory syncytial virus; Role; Severities; Signal Transduction; Supplementation; System; TNF gene; Techniques; Testing; Training; Umbilical Cord Blood; Viral; Viral Pathogenesis; Virus; Virus Diseases; airway epithelium; bead chip; career; career development; cytokine; environmental enrichment for laboratory animals; epigenome; experience; fetal; genomic data; human model; immune activation; immune function; imprint; improved; in utero; in vitro Model; infant infection; infection risk; inflammatory milieu; innate immune function; innovation; mortality; peer; perforin; prenatal exposure; response; seroconversion; single-cell RNA sequencing; skills; transcriptome; transcriptomics; virology

PROJECT SUMMARY/ABSTRACT HIV-exposed, uninfected (HEU) infants experience significantly higher rates of morbidity and mortality due to respiratory syncytial virus (RSV) infection compared to HIV-unexposed (HUU) infants. Natural killer (NK) cells and antigen-presenting cells (APCs) are innate immune cells that play a critical role in controlling RSV infection. We have previously identified abnormalities in NK cells and APCs from HEU infants, but it is not clear whether these abnormalities explain the increased severity of RSV disease observed in this population. This research seeks to address that knowledge gap and will also investigate which in utero exposures are responsible for immune dysfunction in HEU infants. This research is relevant to the mission of the NICHD because it will advance the understanding of immune cross-talk between the pregnant woman and fetus by studying the impact of maternal HIV infection on neonatal innate immune function. The central hypothesis of this proposal is that in utero exposure to the inflammatory environment associated with maternal HIV infection induces DNA methylation changes in innate immune cells that alter NK cell and APC function, and ultimately impair the response to RSV infection. This hypothesis will be tested through the following specific aims: 1) To compare the innate immune response to RSV infection between HEU and HUU infants using an in vitro model of human respiratory infection; 2) To determine the effect of exposure to an environment enriched in inflammatory cytokines on neonatal innate immune cell function; 3) To identify differences in DNA methylation and RNA expression in NK cells and APCs between HEU and HUU infants and determine the effect of in vitro cytokine exposure on the epigenetic and transcriptomic profile of these cells. Aim 1 will be investigated using an innovative respiratory epithelial and endothelial co-culture system along with HEU and HUU cord or peripheral blood mononuclear cells. In Aim 2, HUU NK cells and APCs will be incubated with inflammatory cytokines to simulate HEU infants’ in utero conditions. Aim 3 will generate the first description of the epigenome and transcriptome of HEU compared with HUU infants, using robust techniques including DNA methylation arrays and single cell RNA sequencing. This approach is innovative because it: 1) allows the first investigation of the impact of HEU immune dysregulation on RSV pathogenesis, and 2) may identify the mechanism for immune dysregulation in HEU infants. This project is significant because it has potential to improve health outcomes for the more than 1 million HEU infants born each year. Complimentary to the proposed research plan, a five-year mentored career development training plan has been devised that incorporates didactic learning in genomic data analysis and hands-on training in virology and immunology laboratory skills. The candidate is co-mentored by internationally recognized experts in the fields of virology, immunology, and genomics/bioinformatics. The candidate’s long- term career goal is to become an independent investigator studying the immune effects of HIV exposure.

项目摘要/摘要 艾滋病毒暴露，未感染的（HEU）婴儿的发病率和死亡率明显更高 与艾滋病毒无暴露（HUU）婴儿相比，呼吸综合病毒（RSV）感染。天然杀手（NK）细胞 和抗原呈递细胞（APC）是先天免疫小球，在控制RSV感染中起着至关重要的作用。 我们以前已经从HEU婴儿中发现了NK细胞和APC的异常，但尚不清楚是否清楚是否清楚 这些异常解释了该人群中观察到的RSV疾病严重程度的增加。这项研究 试图解决该知识差距，还将调查子宫暴露中的哪些 HEU婴儿的免疫功能障碍。这项研究与NICHD的使命相关，因为它将进步 通过研究孕妇和胎儿之间免疫串扰的理解 孕产妇HIV感染对新生儿先天免疫功能。 该提议的核心假设是，在子宫内暴露于相关的炎症环境中 孕产妇感染可诱导改变NK细胞和APC的先天免疫细胞的DNA甲基化变化 功能，最终会损害对RSV感染的反应。该假设将通过以下来检验 具体目的：1）比较使用HEU和HUU婴儿之间对RSV感染的先天免疫反应 人类呼吸道感染的体外模型； 2）确定暴露于环境的影响 富含关于新生儿先天免疫功能的炎性细胞因子； 3）确定DNA的差异 HEU和HUU婴儿之间NK细胞和APC中的甲基化和RNA表达，并确定该作用 这些细胞的表观遗传和转录组谱上的体外细胞因子暴露。 AIM 1将被调查 使用创新的呼吸道上皮和内皮共培养系统以及Heu和Huu Cord或 外周血单核细胞。在AIM 2中，Huu NK细胞和APC将与炎症一起孵育 细胞因子在子宫疾病中模拟HEU婴儿的模拟。 AIM 3将产生表观基因组的第一个描述 与HUU婴儿相比，HEU的转录组和使用包括DNA甲基化的强大技术 阵列和单细胞RNA测序。 这种方法具有创新性，因为它：1）允许首次投资HEU免疫的影响 RSV发病机理的失调，2）可以确定HEU免疫失调的机制 婴儿。该项目很重要，因为它有可能改善超过100万的健康状况 HEU婴儿每年出生。拟议的研究计划的补充是一项五年的指导职业 已经设计了开发培训计划，将教学学习纳入基因组数据分析和 病毒学和免疫学实验室技能的动手培训。候选人由国际上 公认的病毒学，免疫学和基因组学/生物信息学领域的专家。候选人的长期 任期职业目标是成为研究艾滋病毒接触的免疫学家影响的独立研究者。