Mountain Laboratories, Laboratory of Bacteriology, National Institute of Allergy and Infectious Diseases, NIH, Hamilton, MT, USA.
Methods Mol Biol. 2020;2136:367-375. doi: 10.1007/978-1-0716-0467-0_29.
Group A Streptococcus (GAS) produces the pore-forming toxin, streptolysin O (SLO). SLO sequesters cholesterol and induces a plasma membrane repair process that removes the pores via a lipid raft-mediated endocytosis. The impact SLO has on membranes makes it an effective toxin for investigating the function of lipid rafts in cellular processes. Lipid rafts are essential for B-cell activation. Indeed, antigen-stimulated B-cell receptors (BCRs) require localization with lipid rafts for efficient signaling and internalization. SLO treatment impairs BCR activation by competing for lipid rafts. Here, disrupting lipid rafts using SLO and assessing the effects on BCR activation by fluorescence microscopy and flow cytometry are described.
A 组链球菌（GAS）产生穿孔毒素，即链球菌溶血素 O（SLO）。SLO 会隔离胆固醇并诱导质膜修复过程，通过脂筏介导的内吞作用去除孔。SLO 对膜的影响使其成为研究脂筏在细胞过程中的功能的有效毒素。脂筏对于 B 细胞的激活是必不可少的。事实上，抗原刺激的 B 细胞受体（BCR）需要与脂筏定位以实现有效的信号转导和内化。SLO 通过与脂筏竞争来干扰 BCR 的激活。本文描述了使用 SLO 破坏脂筏并通过荧光显微镜和流式细胞术评估对 BCR 激活的影响。
