Golightly L M, Thomas L L, Dvorak A M, Ackerman S J
Department of Medicine, Beth Israel Hospital, Boston, Massachusetts.
J Leukoc Biol. 1992 Apr;51(4):386-92. doi: 10.1002/jlb.51.4.386.
The Charcot-Leyden crystal (CLC) protein, a prominent cell constituent unique to eosinophils and basophils, possesses lysophospholipase activity. This activity and the extracellular deposition and formation of CLC in tissues and body fluids in association with eosinophils suggest an extracellular function for this protein in inflammation. During degranulation, basophils release granule-derived mediators of inflammation. We postulated that CLC protein, localized in part to the basophil granule, might be released along with other mediators during this process. The extracellular release of CLC protein was studied during the degranulation of basophils stimulated by anti-immunoglobulin E (anti-IgE), N-formyl-methionyl-leucyl-phenylalanine (fMLP), phorbol myristate acetate, eosinophil major basic protein (MBP), and calcium ionophore A23187. Histamine release was used as a marker of basophil degranulation; its release was measured utilizing the fluorometric technique. CLC protein was not released into the supernatant during this process as determined by radioimmunoassay. CLC protein in the extracellular space, either as intact crystals or aggregates, was undetectable by indirect immunofluorescent staining of basophils activated with either anti-IgE or fMLP. However, upon activation, the immunofluorescent cytoplasmic and nuclear staining pattern for CLC protein was significantly altered. Decreased cytoplasmic staining and persistent or increased nuclear staining for CLC protein were observed after activation, with recovery of the preactivation, unstimulated cellular staining pattern at 30 and 45 min after stimulation with fMLP and anti-IgE, respectively. These findings suggest that CLC protein functions intracellularly in basophils during the process of activation, degranulation, and recovery. The potential nuclear function(s) of this lysophospholipase in the basophil requires further investigation.
夏科-莱登结晶（CLC）蛋白是嗜酸性粒细胞和嗜碱性粒细胞特有的一种重要细胞成分，具有溶血磷脂酶活性。这种活性以及CLC在组织和体液中与嗜酸性粒细胞相关的细胞外沉积和形成表明该蛋白在炎症中具有细胞外功能。在脱颗粒过程中，嗜碱性粒细胞释放颗粒衍生的炎症介质。我们推测，部分定位于嗜碱性粒细胞颗粒的CLC蛋白可能在此过程中与其他介质一起释放。在抗免疫球蛋白E（抗IgE）、N-甲酰甲硫氨酰亮氨酰苯丙氨酸（fMLP）、佛波酯、嗜酸性粒细胞主要碱性蛋白（MBP）和钙离子载体A23187刺激嗜碱性粒细胞脱颗粒过程中，研究了CLC蛋白的细胞外释放。组胺释放用作嗜碱性粒细胞脱颗粒的标志物；利用荧光测定技术测量其释放量。通过放射免疫测定法确定，在此过程中CLC蛋白未释放到上清液中。通过抗IgE或fMLP激活的嗜碱性粒细胞的间接免疫荧光染色未检测到细胞外空间中作为完整晶体或聚集体的CLC蛋白。然而，激活后，CLC蛋白的免疫荧光细胞质和细胞核染色模式发生了显著变化。激活后观察到CLC蛋白的细胞质染色减少，细胞核染色持续或增加，在用fMLP和抗IgE刺激后分别在30分钟和45分钟恢复到激活前未刺激的细胞染色模式。这些发现表明，CLC蛋白在嗜碱性粒细胞激活、脱颗粒和恢复过程中在细胞内发挥作用。这种溶血磷脂酶在嗜碱性粒细胞中的潜在核功能需要进一步研究。
