B-lymphocytes have the ability to repair their plasma membranes following injury, such as by bacterial cholesterol-dependent cytolysins. The repair process includes the removal of the pore from the inflicted region of the plasma membrane via lipid raft-mediated internalization. Lipid rafts are critical for B cell receptor (BCR) activation. Cholesterol-dependent pore forming bacterial toxins provide a useful tool for examining the role of lipid rafts in B cell activation and the underlying cellular mechanisms. This method serves as a great alternative of known cholesterol disruption reagents such as filipin, nystatin, and methyl-β-cyclodextrin. Here, we describe a method of damaging primary murine B cell plasma membranes with the Streptococcus pyogenes cytolysin, Streptolysin O (SLO), and monitoring levels of damage, repair and BCR activation.
CITATION STYLE
Miller, H., & Song, W. (2018). Use of streptolysin O-induced membrane damage as a method of studying the function of lipid rafts during B cell activation. In Methods in Molecular Biology (Vol. 1707, pp. 235–241). Humana Press Inc. https://doi.org/10.1007/978-1-4939-7474-0_17
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