通过流式细胞术对 HIV gp140 进行连续染色，以捕获抗原特异性的人类 B 细胞。

U.S. Military HIV Research Program, Center of Infectious Disease Research, Walter Reed Army Institute of Research, Silver Spring, MD 20910, USA.

Henry M. Jackson Foundation for the Advancement of Military Medicine, Inc., Bethesda, MD 20817, USA.

STAR Protoc. 2021 Aug 24;2(3):100771. doi: 10.1016/j.xpro.2021.100771. eCollection 2021 Sep 17.

Protocols for efficient capture of antigen-specific B cells (ASBCs) are useful for understanding pathogen-specific B-cell responses during natural infection or vaccination. Fluorescently labeled tetramerized probes are classically used to capture ASBCs, but many occlude valuable epitopes available for B-cell receptor binding. Here, we describe a bead assay to confirm ASBC receptor accessibility on probes and a sequential staining process to capture HIV gp140-specific B cells from human peripheral blood mononuclear cells. For complete details on the use and execution of this protocol, please refer to Townsley et al. (2021).

用于高效捕获抗原特异性 B 细胞 (ASBC) 的方案对于理解自然感染或接种疫苗期间针对病原体的 B 细胞反应很有用。荧光标记的四聚体探针常用于捕获 ASBC，但许多探针会掩盖可用于 B 细胞受体结合的有价值的表位。在这里，我们描述了一种珠子测定法来确认探针上的 ASBC 受体可及性，以及一种顺序染色过程，用于从人外周血单核细胞中捕获 HIV gp140 特异性 B 细胞。有关该方案使用和执行的完整详细信息，请参阅 Townsley 等人。(2021)。