Department of Biochemistry and Molecular Biology, Monash University, Clayton, VIC, Australia.
Immunity Program, Biomedicine Discovery Institute, Monash University, Clayton, VIC, Australia.
Methods Mol Biol. 2024;2826:117-129. doi: 10.1007/978-1-0716-3950-4_9.
Memory B cells are central to the establishment of immunological memory, providing long-term protection against specific pathogens and playing a vital role in the efficacy of vaccines. Understanding how memory B cell formation is disrupted during persistent infection is essential for new therapeutics. Lymphocytic choriomeningitis virus (LCMV) is an ideal model for investigating memory B cells in acute versus chronic infection. This protocol details techniques to isolate, enrich, and examine LCMV-specific memory B cells in both acute and chronic LCMV infection. Using an antigen tetramer enrichment system and flow cytometry, this method assesses low-frequency, polyclonal antigen-specific memory B cells.
记忆 B 细胞是建立免疫记忆的核心,为针对特定病原体提供长期保护,并在疫苗的功效中发挥重要作用。了解持续性感染过程中记忆 B 细胞的形成如何受到破坏对于新的治疗方法至关重要。淋巴细胞性脉络丛脑膜炎病毒 (LCMV) 是研究急性和慢性感染中记忆 B 细胞的理想模型。该方案详细介绍了在急性和慢性 LCMV 感染中分离、富集和检查 LCMV 特异性记忆 B 细胞的技术。该方法使用抗原四聚体富集系统和流式细胞术来评估低频、多克隆抗原特异性记忆 B 细胞。