Data_Sheet_1_Clinical Protocol for a Longitudinal Cohort Study Employing Systems Biology to Identify Markers of Vaccine Immunogenicity in Newborn Infants in The Gambia and Papua New Guinea.docx

Background: Infection contributes to significant morbidity and mortality particularly in the very young and in low- and middle-income countries. While vaccines are a highly cost-effective tool against infectious disease little is known regarding the cellular and molecular pathways by which vaccines induce protection at an early age. Immunity is distinct in early life and greater precision is required in our understanding of mechanisms of early life protection to inform development of new pediatric vaccines.
Methods and Analysis: We will apply transcriptomic, proteomic, metabolomic, multiplex cytokine/chemokine, adenosine deaminase, and flow cytometry immune cell phenotyping to delineate early cellular and molecular signatures that correspond to vaccine immunogenicity. This approach will be applied to a neonatal cohort in The Gambia (N ~ 720) receiving at birth: (1) Hepatitis B (HepB) vaccine alone, (2) Bacille Calmette Guerin (BCG) vaccine alone, or (3) HepB and BCG vaccines, (4) HepB and BCG vaccines delayed till day 10 at the latest. Each study participant will have a baseline peripheral blood sample drawn at DOL0 and a second blood sample at DOL1,−3, or−7 as well as late timepoints to assess HepB vaccine immunogenicity. Blood will be fractionated via a “small sample big data” standard operating procedure that enables multiple downstream systems biology assays. We will apply both univariate and multivariate frameworks and multi-OMIC data integration to identify features associated with anti-Hepatitis B (anti-HB) titer, an established correlate of protection. Cord blood sample collection from a subset of participants will enable human in vitro modeling to test mechanistic hypotheses identified in silico regarding vaccine action. Maternal anti-HB titer and the infant microbiome will also be correlated with our findings which will be validated in a smaller cohort in Papua New Guinea (N ~ 80).
Ethics and Dissemination: The study has been approved by The Gambia Government/MRCG Joint Ethics Committee and The Boston Children's Hospital Institutional Review Board. Ethics review is ongoing with the Papua New Guinea Medical Research Advisory Committee. All de-identified data will be uploaded to public repositories following submission of study output for publication. Feedback meetings will be organized to disseminate output to the study communities.
Clinical Trial Registration: Clinicaltrials.gov Registration Number: NCT03246230

研究背景：感染可导致显著的发病率与死亡率，尤其在极幼龄人群以及中低收入国家中。尽管疫苗是对抗感染性疾病的高成本效益工具，但目前对于疫苗在生命早期诱导保护作用的细胞与分子通路仍知之甚少。生命早期的免疫状态具有独特性，因此需要更精准地阐明早期生命保护机制，为新型儿童疫苗的开发提供依据。

研究方法与分析：本研究将采用转录组学（transcriptomic）、蛋白质组学（proteomic）、代谢组学（metabolomic）、多重细胞因子/趋化因子检测、腺苷脱氨酶检测以及流式细胞术免疫细胞表型分析等手段，阐明与疫苗免疫原性对应的早期细胞与分子特征谱。本研究将对冈比亚的新生儿队列（样本量约720例）实施上述分析策略，该队列新生儿出生时分别接受以下4种疫苗接种方案：(1) 单独接种乙型肝炎（HepB）疫苗；(2) 单独接种卡介苗（BCG）；(3) 同时接种乙型肝炎与卡介苗；(4) 乙型肝炎与卡介苗接种最迟延迟至出生后第10天。所有研究参与者均在出生后第0天采集外周血基线样本，并在出生后第1、3或7天以及晚期随访时间点采集第二份血样，以评估乙型肝炎疫苗的免疫原性。血液样本将通过“小样本大数据”标准操作规程进行分离，可支持多种下游系统生物学检测。本研究将采用单变量与多变量分析框架以及多组学数据整合策略，识别与抗乙型肝炎（抗HB）滴度（已确立的保护相关标志物）相关的特征。对部分参与者采集的脐带血样本将用于开展体外模型实验，以验证通过计算机模拟（in silico）分析得出的疫苗作用机制假说。此外，本研究还将分析母体抗HB滴度、婴儿微生物组与本研究结果的相关性，并在巴布亚新几内亚的小型队列（样本量约80例）中对研究发现进行验证。

伦理与成果传播：本研究已获得冈比亚政府/冈比亚医学研究委员会（MRCG）联合伦理委员会以及波士顿儿童医院机构审查委员会的批准。巴布亚新几内亚医学研究咨询委员会的伦理审查工作正在进行中。所有去标识化数据将在研究成果投稿发表后上传至公共数据库。本研究将组织反馈会议，向研究社区分享研究成果。

临床试验注册：美国临床试验数据库（Clinicaltrials.gov）注册编号：NCT03246230