A longitudinal immunological and virological study for ME CFS biomarker discovery (Renewal)

这是RO 1研究“ME/CFS生物标志物的纵向免疫学研究”的更新申请 发现”。我们将招募和随访新病例以及英国ME/CFS队列的病例样本，重点关注 详细的免疫学和临床表型，并分析其相互关系。 受试者和纵向评估：110例ME/CFS病例（1/2重度，1/2轻度-中度）符合所有 CDC-19942、CIM 1和IOM 4标准将每6至12个月进行一次评估，共5个时间点， 将生物标志物表达的变化映射到临床参数上（反之亦然）;精确分层 不同的病例类型;并准确分析疾病进展不同阶段的生物标志物及其 与临床参数的时间关系。 活动和目标：在一项前瞻性队列研究中，我们将：i）招募并随访共计110名ME/CFS 病例，其中至少100例病例将有完整的数据和样本集; ii）根据以下因素对患者进行分层： 症状严重程度的趋势，a）“改善”，B）“稳定”或c）“恶化”，使用与疼痛、疲劳和 功能状态39.对于深入的免疫学分析，我们将：i）进行详细的离体表型分析， 通过流式细胞术测量PBMC群体; ii）测量体外培养后NK和T细胞的功能应答; 通过流式细胞术刺激; iii）通过流式细胞术分析刺激的PBMC培养物上清液中分泌的细胞因子， 多重微珠阵列;和iv）MHC 1类和KIR的基因型供体，以告知流式细胞术分析 数据整合分析并将免疫学数据映射到临床数据中，并确定 免疫参数先于、跟随或预测临床轨迹，我们将：i）量化 免疫学生物标志物和ii）鉴定与ME/CFS临床状态变化相关的生物标志物。我们 将在各个阶段考虑患者的观点、道德、法律的、社会问题。 为了确保质量和最大的研究产出，我们将继续使用英国ME/CFS队列程序， 在患者的纵向随访过程中。我们将遵循标准化的临床、数据和样本 符合特定临床标准的ME/CFS患者的随访中的处理和实验室程序。 创新和战略需求：据我们所知，这是ME/CFS的第一个长期前瞻性研究， 包括门诊和重症病例，并包括全面的临床数据和深入的 免疫学分析这项研究的独特之处在于其严格的入选标准和详细的临床表型， 为我们的分析增加了特异性和有效性。从长远来看，确定可靠的生物标志物将使 ME/CFS表型与疾病严重程度和预后的相关性，并可能揭示新的选择， 干预措施。由于1- 250万美国人患有ME/CFS 4，这项研究有可能改善 改善美国大量患者的生活，并推动美国该领域的发展， 化国际大患者和利益相关者参与了研究的各个方面。

This is a renewal application for the RO1 study ‘A longitudinal immunological study for ME/CFS biomarker discovery’. We will recruit and follow-up new cases and a sample of cases of the UK ME/CFS cohort, focusing on detailed immunological and clinical phenotypes and analyses of their inter-relationships. Participants and longitudinal assessments: 110 ME/CFS cases (½ severe, ½ mild-moderate) meeting all of the CDC-19942, Canadian1 and IOM4 criteria will be assessed every 6 to 12 months for 5 time-points, enabling the mapping of changes in biomarker expression onto clinical parameters (and vice versa); to precisely stratify different case types; and to accurately analyse biomarkers at different stages of disease progression and their time-relationship with clinical parameters. Activities and objectives: In a prospective cohort study, we will: i) recruit and follow up a total of 110 ME/CFS cases, of which at least 100 cases will have complete data and sample sets; ii) stratify patients according to trends in symptom severity, a) ‘improving’, b) ‘stable’ or c) ‘worsening’, using scores related to pain, fatigue and functional status39. For in-depth immunological profiling, we will: i) conduct detailed ex vivo phenotyping of PBMC populations by flow cytometry; ii) measure the functional response of NK and T cells after in vitro stimulation by flow cytometry; iii) analyze secreted cytokines in supernatants of stimulated PBMC cultures by multiplex bead array; and, iv) genotype donors for MHC Class1 and KIR to inform analysis of flow cytometry data. To integrate analyses and map immunological data onto clinical data and determine whether changes in immune parameters precede, follow, or predict the clinical trajectory, we will: i) quantify correlations between immunological biomarkers and ii) identify biomarkers associated with changes in ME/CFS clinical status. We will consider patients’ perspectives, ethical, legal, societal issues at all stages. To ensure quality and maximum research output we will continue using the UK ME/CFS cohort procedures and protocols during the longitudinal follow up of patients. We will follow standardised clinical, data and samples handling and laboratory procedures in the follow up of ME/CFS patients meeting specific clinical criteria. Innovation and Strategic Need: This is, to our knowledge, the first long-term prospective study of ME/CFS to incorporate both ambulatory and severe cases and to include comprehensive clinical data and in-depth immunological profiling. The study is unique for its strict inclusion criteria and detailed clinical phenotyping, adding specificity and validity to our analyses. In the long term, identification of robust biomarkers will allow the correlation of ME/CFS phenotype with disease severity and prognosis and may reveal new options for interventions. Because 1- 2.5 million Americans have ME/CFS4, this study has the potential to improve the lives of a large patient population in the U.S. as well as advance the state of the field in the U.S. and internationally. Patients and stakeholders are involved in all aspects of the research.