Lehmann A K, Halstensen A, Holst J, Bassøe C F
Medical Department B, University of Bergen, Haukeland Hospital, Norway.
J Immunol Methods. 1997 Jan 15;200(1-2):55-68. doi: 10.1016/s0022-1759(96)00185-8.
Functional flow cytometry and chemiluminescence (CL) assays have been modified to identify serogroup B meningococcal structures that mediate anti-meningococcal opsonophagocytosis. Serogroup B meningococcal outer membrane vesicles (OMV) were adsorbed to fluorescent latex beads (OMV-beads) and opsonized with acute phase and convalescence sera from patients with serogroup B meningococcal disease. Phagocytosis of these beads by human monocytes and polymorphonuclear leukocytes (non-lymphocytes) was dependent on both antigen exposure on the bead surface and on serum opsonization. OMV-beads opsonized with serum from a patient recovering from meningococcal disease, caused 97% of the non-lymphocytes to phagocytose an average of 15.8 beads per cell with a CL response of 46,550 mVs, whereas opsonized control beads were phagocytosed by 19% of the non-lymphocytes with 1.1 beads per cell and a CL response of 53 mVs. Increased amounts of functional, anti-OMV opsonins were detected during infection, and opsonized OMV-beads elicited phagocyte responses of similar magnitude to those of opsonized whole meningococci. Phagocyte internalization of OMV-beads was confirmed by confocal laser scanning microscopy. We conclude that epitopes on the meningococcal outer membrane are recognized by anti-meningococcal opsonins in these functional phagocytosis assays, which provide a basis for subsequent evaluation of various purified bacterial components as mediators of human opsonophagocytic responses and hence future vaccine constituents.
功能性流式细胞术和化学发光（CL）分析已被改进，以鉴定介导抗脑膜炎球菌调理吞噬作用的B群脑膜炎球菌结构。将B群脑膜炎球菌外膜囊泡（OMV）吸附到荧光乳胶珠（OMV-珠）上，并用B群脑膜炎球菌病患者的急性期和恢复期血清进行调理。人单核细胞和多形核白细胞（非淋巴细胞）对这些珠子的吞噬作用既取决于珠子表面的抗原暴露，也取决于血清调理作用。用从脑膜炎球菌病康复患者的血清调理的OMV-珠，使97%的非淋巴细胞吞噬，每个细胞平均吞噬15.8个珠子，化学发光反应为46,550 mVs，而调理的对照珠被19%的非淋巴细胞吞噬，每个细胞吞噬1.1个珠子，化学发光反应为53 mVs。在感染期间检测到功能性抗OMV调理素的量增加，调理的OMV-珠引发的吞噬细胞反应与调理的完整脑膜炎球菌的反应大小相似。通过共聚焦激光扫描显微镜证实了OMV-珠被吞噬细胞内化。我们得出结论，在这些功能性吞噬分析中，脑膜炎球菌外膜上的表位被抗脑膜炎球菌调理素识别，这为随后评估各种纯化的细菌成分作为人类调理吞噬反应的介质以及未来疫苗成分提供了基础。
