Laboratoire Colloïdes et Matériaux Divisés (LCMD), ESPCI Paris, PSL Research University, CNRS UMR8231 Chimie Biologie Innovation, Paris, France.
Laboratoire de Biochimie (LBC), ESPCI Paris, PSL Research University, CNRS UMR8231 Chimie Biologie Innovation, Paris, France.
Nat Biotechnol. 2017 Oct;35(10):977-982. doi: 10.1038/nbt.3964. Epub 2017 Sep 11.
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. 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.
对抗体介导的免疫反应动力学的研究一直受到缺乏定量、高通量系统来分析单个抗体分泌细胞的阻碍。在这里，我们描述了一种简单的微流控系统 DropMap，其中单个细胞被分隔在成千上万的 40 微升液滴中，并在二维液滴阵列中使用荧光重定位免疫测定法进行分析。使用 DropMap，我们在破伤风类毒素 (TT) 免疫的小鼠中进行了为期 7 周的方案，同时分析了从脾脏和骨髓中富集的超过 50 万个单个细胞中 IgG 的分泌率和亲和力。免疫接种导致单个细胞分泌率和亲和力的范围都显著增加，分别最大增加了 3 和 4 个对数。我们观察到在时间内，在解剖隔室内部和之间，分泌率和亲和力的动力学存在差异。该系统不仅将能够进行免疫监测和优化免疫接种和疫苗接种方案，还将增强抗体筛选。
