High voltage immunoelectron microscopy of complement receptor type 3-mediated capping and internalization of group A streptococcal cell walls by human neutrophils.
High voltage immunoelectron microscopy of complement receptor type 3-mediated capping and internalization of group A streptococcal cell walls by human neutrophils.
复制标题
高压免疫电子显微镜观察补体受体 3 型介导的人中性粒细胞对 A 组链球菌细胞壁的加帽和内化。
DOI：
10.1002/jemt.1070280403
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发表时间：
1994
影响因子：
2.5
通讯作者：
Pryzwansky,KB
中科院分区：
文献类型：
--
作者：
Pryzwansky,KB
通过高压免疫电镜（HVEM）研究了人类中性粒细胞清除源自链球菌细胞壁（PG‐APS）的肽聚糖a组特异性多糖聚合物的机制，以确定中性粒细胞如何处理这种高度炎症性的细菌碎片。中性粒细胞单层与血清活化的PG - APS孵育5-30分钟。用0.5%戊二醛轻轻固定细胞，用N -乙酰氨基葡萄糖抗体和15 nm胶体金偶联山羊抗兔IgG，免疫金将PG - APS定位在中性粒细胞表面。用立体HVEM观察未切片的中性粒细胞。PG - APS在5分钟内随机分布在分布良好的中性粒细胞的质膜上。在极化细胞中，PG‐APS密集分布于尾足纤维和收缩纤维上。在15分钟内，PG‐APS主要集中成一个大的聚集体，直径约为1 μm，靠近细胞边缘或细胞核。PG‐APS的聚集体被吞没在细胞核压痕附近（hof）。PG - APS的聚集和内化需要完整的微丝。PG‐APS的结合依赖于补体固定。此外，通过免疫金标记抗CR3的形态测定，PG‐APS诱导中性粒细胞表面补体受体3 （CR3， C3bi受体）的密度增加。当对细胞进行PG - APS和CR3染色时，观察到共定位，体视显微镜显示与吞噬相关的区域有CR3簇。这些数据表明，中性粒细胞使用一种有效的机制来清除细菌碎片。与整个链球菌在多个部位被吞噬不同，这些细菌细胞壁首先被收集成一个大的聚集体或帽，然后在一个部位被内化。©1994 Wiley‐Liss, Inc。
The mechanism of human neutrophil clearance of peptidoglycan group A–specific polysaccharide polymers derived from streptococcal cell walls (PG‐APS) was investigated by high voltage immunoelectron microscopy (HVEM) in order to determine how neutrophils process this highly inflammatory bacterial debris. Neutrophil monolayers were incubated from 5–30 min with serum‐opsonized PG‐APS. Cells were lightly fixed with 0.5% glutaraldehyde, and the PG‐APS was localized on the neutrophil surface by immunogold using antibodies to N‐acetyl‐glucosamine and 15 nm colloidal gold coupled to goat anti‐rabbit IgG. Neutrophils were viewed unsectioned by stereo HVEM. Patches of PG‐APS were distributed randomly on the plasmalemma of well‐spread neutrophils within 5 min. In polarized cells, PG‐APS was densely localized on the uropod and retraction fibers. Within 15 min, PG‐APS was predominantly concentrated into a large aggregate, measuring approximately 1 μm in diameter, near the cell margin or nucleus. The aggregate of PG‐APS was engulfed in the vicinity of the indentation of the nucleus (hof). Intact microfilaments were required for aggregation and internalization of PG‐APS. Binding of PG‐APS was dependent upon complement fixation. Furthermore, PG‐APS elicited an increase in density of complement receptor type 3 (CR3, C3bi receptor) on the neutrophil surface as determined by morphometry of immunogold labeled anti‐CR3. When cells were stained for both PG‐APS and CR3, co‐localization was observed, and stereomicroscopy revealed clusters of CR3 in areas associated with phagocytosis. These data suggest that neutrophils use an efficient mechanism for removal of bacterial debris. Unlike whole streptococci which are phagocytosed at multiple sites, these bacterial cell walls are first collected into a large aggregate, or cap, which is then internalized at one site. © 1994 Wiley‐Liss, Inc.