Department of Microbiology and Immunology, University at Buffalo School of Medicine, Buffalo, NY, USA.
Methods Mol Biol. 2021;2183:559-574. doi: 10.1007/978-1-0716-0795-4_33.
Antibodies against Streptococcus pneumoniae (pneumococcus) following vaccination are crucial for host protection against invasive pneumococcal infections. The antibodies induced by pneumococcal vaccines act as opsonins to mediate bacterial uptake and killing by host phagocytic cells, especially polymorphonuclear leukocytes (PMNs) also called neutrophils. Therefore, it is important to measure not only the levels of antibodies induced by a pneumococcal vaccine candidate but their actual functional capacity in mediating bacterial opsonization and killing by PMNs. Here, we describe a protocol to demonstrate effective deposition of vaccine-induced antibodies on the surface of S. pneumoniae by flow cytometry and subsequent opsonophagocytic killing (OPH) by murine bone-marrow derived PMNs.
接种疫苗后针对肺炎链球菌(肺炎球菌)的抗体是宿主预防侵袭性肺炎球菌感染的关键。肺炎球菌疫苗诱导的抗体作为调理素,介导宿主吞噬细胞(特别是多形核白细胞(PMN),也称为中性粒细胞)摄取和杀死细菌。因此,重要的是不仅要测量肺炎球菌候选疫苗诱导的抗体水平,还要测量其介导 PMN 调理细菌吞噬和杀伤的实际功能能力。在这里,我们描述了一种通过流式细胞术证明疫苗诱导的抗体有效沉积在肺炎链球菌表面,随后通过鼠骨髓衍生的 PMN 调理吞噬杀伤(OPH)的方案。