Newman S L, Gootee L, Gabay J E
Department of Medicine, University of Cincinnati College of Medicine, Ohio 45267.
J Clin Invest. 1993 Aug;92(2):624-31. doi: 10.1172/JCI116630.
Human neutrophils (PMN) demonstrated potent fungistatic activity against Histoplasma capsulatum (Hc) yeasts in a sensitive microassay that quantifies the growth of yeasts by the incorporation of [3H]leucine. At a PMN:yeast ratio of 1:2, PMN inhibited the growth of yeasts by 37%. Maximum inhibition of 85% to 95% was achieved at a PMN/yeast ratio of 10:1 to 50:1. Opsonization of the yeasts in fresh or heat-inactivated serum was required for PMN-mediated fungistasis, but ingestion of the yeasts was not required. Recognition and phagocytosis of opsonized yeasts was via PMN complement receptor (CR) type 1 (CR1), CR3, and FcRIII (CD16). PMN fungistatic activity was evident by 2 h, was maximum at 24 h, and persisted up to 5 d. In contrast, yeasts multiplied within monocytes to a greater extent than in culture medium alone. PMN from three patients with chronic granulomatous disease (CGD) inhibited the growth of Hc yeasts by an average of 97%, compared with 86% in three normal controls. Furthermore, preincubation of PMN with the lysosomotropic agent NH4Cl inhibited fungistatic activity in a concentration-dependent manner. Finally, experiments with subcellular fractions of PMN demonstrated that the principal component of the fungistatic activity of PMN was localized in the azurophil granules. These data demonstrate that human PMN possess potent fungistatic activity against Hc yeasts and further show that fungistasis is mediated by antimicrobial agents contained in the azurophil granules.
在一项灵敏的微量测定中，人类中性粒细胞（PMN）对荚膜组织胞浆菌（Hc）酵母表现出强大的抑菌活性，该测定通过掺入[3H]亮氨酸来量化酵母的生长。在PMN与酵母比例为1:2时，PMN抑制酵母生长37%。在PMN/酵母比例为10:1至50:1时，实现了85%至95%的最大抑制率。PMN介导的抑菌作用需要新鲜或热灭活血清中酵母的调理作用，但不需要吞噬酵母。调理后的酵母通过PMN补体受体（CR）1型（CR1）、CR3和FcRIII（CD16）被识别和吞噬。PMN的抑菌活性在2小时时明显，在24小时时达到最大值，并持续长达5天。相比之下，酵母在单核细胞内的增殖程度比仅在培养基中更大。三名慢性肉芽肿病（CGD）患者的PMN平均抑制Hc酵母生长97%，而三名正常对照为86%。此外，用溶酶体促渗剂NH4Cl预孵育PMN以浓度依赖的方式抑制抑菌活性。最后，对PMN亚细胞组分的实验表明，PMN抑菌活性的主要成分定位于嗜天青颗粒中。这些数据表明，人类PMN对Hc酵母具有强大的抑菌活性，并进一步表明抑菌作用是由嗜天青颗粒中含有的抗菌剂介导的。
