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.

项目总结/摘要 暴露于艾滋病毒的未感染婴儿的发病率和死亡率明显较高， 呼吸道合胞病毒（RSV）感染与HIV未暴露（HUU）婴儿相比。自然杀伤（NK）细胞 而抗原呈递细胞（APC）是在控制RSV感染中起关键作用的先天免疫细胞。 我们以前已经发现高浓缩铀婴儿的NK细胞和APC异常，但尚不清楚是否 这些异常解释了在这一人群中观察到的RSV疾病严重程度的增加。本研究 旨在解决这一知识差距，并将调查子宫内暴露是造成 高浓缩铀婴儿的免疫功能障碍。这项研究与NICHD的使命有关，因为它将推动 通过研究孕妇和胎儿之间的免疫串扰的影响， 母亲HIV感染对新生儿先天免疫功能的影响。 该建议的中心假设是，在子宫内暴露于炎症环境相关 母体HIV感染诱导先天免疫细胞DNA甲基化改变，改变NK细胞和APC 功能，并最终损害对RSV感染的反应。这一假设将通过以下方式进行检验： 具体目的：1）使用免疫学方法比较HEU和HUU婴儿对RSV感染的先天免疫应答。 人呼吸道感染的体外模型; 2）确定暴露于环境的影响 3）为了确定DNA中的差异， HEU和HUU婴儿之间NK细胞和APC的甲基化和RNA表达，并确定其影响 体外细胞因子暴露对这些细胞的表观遗传和转录谱的影响。将研究目标1 使用创新的呼吸道上皮和内皮共培养系统沿着HEU和HUU脐带或 外周血单个核细胞在目标2中，HUU NK细胞和APC将与炎性细胞一起孵育。 细胞因子来模拟高浓缩铀婴儿在子宫内的条件。目标3将产生表观基因组的第一个描述 使用包括DNA甲基化在内的可靠技术， 阵列和单细胞RNA测序。 这一方法具有创新性，因为它： RSV发病机制的调节异常，和2）可以确定HEU中免疫调节异常的机制 婴儿。该项目意义重大，因为它有可能改善100多万人的健康状况。 每年出生的高浓缩铀婴儿。作为对拟议研究计划的补充， 制定了发展培训计划，将教学式学习纳入基因组数据分析， 病毒学和免疫学实验室技能的实践培训。候选人由国际上的 病毒学、免疫学和基因组学/生物信息学领域的公认专家。候选人很长- 我的职业目标是成为一名独立的研究人员，研究艾滋病毒暴露对免疫的影响。