Single-cell deep phenotyping of IgG-secreting cells for high-resolution immune monitoring
Single-cell deep phenotyping of IgG-secreting cells for high-resolution immune monitoring
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DOI：
10.1038/nbt.3964
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发表时间：
2017-10-01
影响因子：
46.9
通讯作者：
Baudry, Jean
中科院分区：
文献类型：
--
作者：
Eyer, Klaus;Doineau, Raphael C. L.;Baudry, Jean
抗体介导的免疫应答的动力学研究由于缺乏定量、高通量系统来分析单个抗体分泌细胞而受到阻碍（1-5）。在这里，我们描述了一个简单的微流控系统，DropMap，其中单个细胞被划分成数万个40 pL的液滴，并在二维液滴阵列中使用基于荧光重定位的免疫分析进行分析。使用DropMap，我们在用破伤风类毒素（TT）免疫的小鼠中表征了7周内的抗体分泌细胞，同时分析了从脾和骨髓中富集的超过50万个细胞的IgG的分泌速率和亲和力。免疫接种导致单细胞分泌率和亲和力的范围急剧增加，分别跨越最大3和4个对数。我们观察到随着时间的推移，分泌速率和亲和力的动态解剖区室内和之间的差异。该系统不仅能够进行免疫监测和优化免疫和疫苗接种方案，还能加强抗体筛选（6，7）。
Studies of the dynamics of the antibody-mediated immune response have been hampered by the absence of quantitative, high-throughput systems to analyze individual antibody-secreting cells(1-5). Here we describe a simple microfluidic system, DropMap, in which single cells are compartmentalized in tens of thousands of 40-pL droplets and analyzed in two-dimensional droplet arrays using a fluorescence relocation-based immunoassay. Using DropMap, we characterized antibody-secreting cells in mice immunized with tetanus toxoid (TT) over a 7-week protocol, simultaneously analyzing the secretion rate and affinity of IgG from over 0.5 million individual cells enriched from spleen and bone marrow. Immunization resulted in dramatic increases in the range of both single-cell secretion rates and affinities, which spanned at maximum 3 and 4 logs, respectively. We observed differences over time in dynamics of secretion rate and affinity within and between anatomical compartments. This system will not only enable immune monitoring and optimization of immunization and vaccination protocols but also potentiate antibody screening(6,7).