Measurement and clinical monitoring of human lymphocyte clonality by massively parallel VDJ pyrosequencing.
Measurement and clinical monitoring of human lymphocyte clonality by massively parallel VDJ pyrosequencing.
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DOI：
10.1126/scitranslmed.3000540
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发表时间：
2009-12-23
影响因子：
17.1
通讯作者：
Fire AZ
中科院分区：
文献类型：
--
作者：
Boyd SD;Marshall EL;Merker JD;Maniar JM;Zhang LN;Sahaf B;Jones CD;Simen BB;Hanczaruk B;Nguyen KD;Nadeau KC;Egholm M;Miklos DB;Zehnder JL;Fire AZ
由B和T细胞产生的免疫受体的复杂库使得能够识别对宿主生物体的不同威胁。在这项工作中，我们表明，重排的免疫受体基因座的大规模并行DNA测序可以提供直接检测和跟踪的免疫多样性和扩大克隆淋巴细胞群体在生理和病理环境。从血液和组织样品中分离DNA，使用一系列冗余引物扩增不同的DNA重排，并使用长读超深测序对所得条形码扩增子的混合物进行测序。然后，基于已经连接以形成功能性（或非功能性）免疫效应物的DNA片段来表征单个DNA分子。目前的实验设计可以在单次测序运行中容纳多达150个样品，测序的深度足以在正常和患病情况下鉴定免疫库的稳定和动态方面。这些数据提供了正常个体和血液恶性肿瘤患者的免疫谱的高分辨率图像，在后一种情况下，阐明了克隆肿瘤群体的初始行为和治疗后这些群体的后期抑制或重新出现。
The complex repertoire of immune receptors generated by B and T cells enables recognition of diverse threats to the host organism. In this work, we show that massively parallel DNA sequencing of rearranged immune receptor loci can provide direct detection and tracking of immune diversity and expanded clonal lymphocyte populations in physiological and pathological contexts. DNA was isolated from blood and tissue samples, a series of redundant primers was used to amplify diverse DNA rearrangements, and the resulting mixtures of barcoded amplicons were sequenced using long-read ultra deep sequencing. Individual DNA molecules were then characterized on the basis of DNA segments that had been joined to make a functional (or nonfunctional) immune effector. Current experimental designs can accommodate up to 150 samples in a single sequence run, with the depth of sequencing sufficient to identify stable and dynamic aspects of the immune repertoire in both normal and diseased circumstances. These data provide a high-resolution picture of immune spectra in normal individuals and in patients with hematological malignancies, illuminating, in the latter case, both the initial behavior of clonal tumor populations and the later suppression or re-emergence of such populations after treatment.
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影响因子：
56.9
作者：
Arstila, TP;Casrouge, A;Kourilsky, P
通讯作者：
Kourilsky, P
影响因子：
158.5
作者：
Lowsky, R;Takahashi, T;Strober, S
通讯作者：
Strober, S
DOI：
10.1073/pnas.0801523105
发表时间：
2008-09-02
影响因子：
11.1
作者：
Campbell, Peter J.;Pleasance, Erin D.;Stratton, Michael R.
通讯作者：
Stratton, Michael R.
影响因子：
14.2
作者：
Lim, Annick;Luderschmidt, Stephan;Mempel, Martin
通讯作者：
Mempel, Martin
影响因子：
5.4
作者：
Pinna, Debora;Corti, Davide;Jarrossay, David;Sallusto, Federica;Lanzavecchia, Antonio
通讯作者：
Lanzavecchia, Antonio