Tumor Microenvironment and Cancer Immunology Program, NCI-Designated Cancer Center, Sanford Burnham Prebys Medical Discovery Institute, La Jolla, CA 92037, USA.
STAR Protoc. 2021 Mar 2;2(1):100369. doi: 10.1016/j.xpro.2021.100369. eCollection 2021 Mar 19.
Humoral immune responses depend on the generation of high-affinity antigen-specific antibodies. Germinal center (GC) B cells are the cornerstone of this response in peripheral lymphoid organs. High purities of GC B cells, and also naive B cells, are required for accurate analysis in downstream assays to yield essential knowledge on immunity. This protocol lays out quick and easy steps to purify GC B cells from spleens of immunized mice or B cells from naive animals by negative selection using MACS. For complete details on the use and execution of this protocol, please refer to Ramezani-Rad et al. (2020).
体液免疫反应依赖于高亲和力抗原特异性抗体的产生。生发中心 (GC) B 细胞是外周淋巴器官中该反应的基石。为了在下游分析中进行准确分析,以获得有关免疫的重要知识,需要高纯度的 GC B 细胞和幼稚 B 细胞。本方案通过 MACS 阴性选择,从免疫小鼠的脾脏或幼稚动物的 B 细胞中快速简便地纯化 GC B 细胞。有关此方案的使用和执行的完整详细信息,请参见 Ramezani-Rad 等人。(2020 年)。