Effective Immunological Guidance of Genetic Analyses Including Exome Sequencing in Patients Evaluated for Hemophagocytic Lymphohistiocytosis
Effective Immunological Guidance of Genetic Analyses Including Exome Sequencing in Patients Evaluated for Hemophagocytic Lymphohistiocytosis
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DOI：
10.1007/s10875-017-0443-1
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发表时间：
2017-11-01
影响因子：
9.1
通讯作者：
Ehl, Stephan
中科院分区：
文献类型：
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作者：
Ammann, Sandra;Lehmberg, Kai;Ehl, Stephan
我们报告我们的经验，在使用流式细胞术为基础的免疫学筛查前瞻性的决策工具，使用遗传学研究的诊断方法与噬血细胞性淋巴组织细胞增生症（HLH）的患者。我们将遗传分析主要限于免疫学筛查异常的患者，但对桑格测序结果正常的患者进行全外显子组测序（WES）。在2010年至2014年期间分析的290名疑似HLH儿童（包括17名受影响但无症状的兄弟姐妹）中，87/162名“完全”HLH患者和79/111名“不完全/非典型”HLH患者的免疫学筛查结果正常。在10例患者中，无法检测脱粒。在166例正常筛查的患者中，107例未进行遗传分析（均随访顺利），而在其余59例患者中通过桑格测序进行的154次单基因检测仅确定了1例非典型CHS患者。流式细胞术正确预测了所有29例FHL-2、XLP 1或2患者。在85例NK细胞脱颗粒缺陷患者（包括13例无症状同胞）中，对70例进行了桑格测序，结果55例（79%）进行了基因诊断。8名患者接受了WES，揭示了两个已知和一个未知的细胞毒性基因和一种代谢性疾病的突变。FHL 3是最常见的基因诊断。免疫学筛查提供了一个很好的决策工具，需要和HLH患者的遗传分析的深度，并提供功能相关的信息，快速患者分类，有助于显着减少从诊断到移植的时间，在最近几年。
We report our experience in using flow cytometry-based immunological screening prospectively as a decision tool for the use of genetic studies in the diagnostic approach to patients with hemophagocytic lymphohistiocytosis (HLH). We restricted genetic analysis largely to patients with abnormal immunological screening, but included whole exome sequencing (WES) for those with normal findings upon Sanger sequencing. Among 290 children with suspected HLH analyzed between 2010 and 2014 (including 17 affected, but asymptomatic siblings), 87/162 patients with "full" HLH and 79/111 patients with "incomplete/atypical" HLH had normal immunological screening results. In 10 patients, degranulation could not be tested. Among the 166 patients with normal screening, genetic analysis was not performed in 107 (all with uneventful follow-up), while 154 single gene tests by Sanger sequencing in the remaining 59 patients only identified a single atypical CHS patient. Flow cytometry correctly predicted all 29 patients with FHL-2, XLP1 or 2. Among 85 patients with defective NK degranulation (including 13 asymptomatic siblings), 70 were Sanger sequenced resulting in a genetic diagnosis in 55 (79%). Eight patients underwent WES, revealing mutations in two known and one unknown cytotoxicity genes and one metabolic disease. FHL3 was the most frequent genetic diagnosis. Immunological screening provided an excellent decision tool for the need and depth of genetic analysis of HLH patients and provided functionally relevant information for rapid patient classification, contributing to a significant reduction in the time from diagnosis to transplantation in recent years.