Daimond R D, Krzesicki R
J Clin Invest. 1978 Feb;61(2):360-9. doi: 10.1172/JCI108946.
Mechanisms were studied that might explain the attachment and damage to Candida albicans pseudohyphae by neutrophils in the absence of serum. Attachment of neutrophils to pseudo hyphae was inhibited by Candida mannans (1-10 mg/ml), but not by mannose, dextran, chitin, conconavalin A, or highly charged polyamino acids. Contact was also inhibited by pretreatment of Candida before incubation with neutrophils with chymotrypsin, but not trypsin or several inhibitors of proteases. Similar results were obtained with pretreatment of neutrophils, except that trypsin was inhibitory. When pseudohyphae were killed with ultraviolet light, proteinpolysaccharide complexes of mol wt <10,000 were released which appeared to bind to the surfaces of neutrophils and inhibit contact between neutrophils and Candida, as well as other fungi. Damage to Candida by neutrophils was inhibited by agents known to act on neutrophil oxidative microbicidal mechanisms, including sodium cyanide, sodium azide, catalase, superoxide dismutase, and 1, 4 diazobicyclo (2, 2, 2) octane, a singlet oxygen quencher. Neutrophils from a patient with chronic granulomatous disease did not damage Candida at all. However, the hydroxyl radical scavengers mannitol and benzoate were not inhibitory. Cationic proteins and lactoferrin also did not appear to play a major role in this system. Low concentrations of lysozyme which did not damage Candida in isotonic buffer solutions damaged pseudohyphae in distilled water. Isolated neutrophil granules damaged pseudohyphae only with added hydrogen peroxide and halide, and damage occurred only with granule fractions known to contain myeloperoxidase. These findings suggest that neutrophils recognized a molecule on the Candida surface which has a chymotrypsin sensitive protein component, and which may be liberated from the cell surface upon death of organism. The neutrophil receptors for Candida appear to be sensitive to trypsin and chymotrypsin. Damage to Candida by neutrophils occurred primarily by oxidative mechanisms, including the production of superoxide and hydrogen peroxide interacting with myeloperoxidase and halide, as well as singlet oxygen, but did not appear to involve hydroxyl radical. Lysozyme might have an accessory role, under some conditions.
对无血清情况下中性粒细胞黏附并损伤白色念珠菌假菌丝的机制进行了研究。念珠菌甘露聚糖（1-10毫克/毫升）可抑制中性粒细胞与假菌丝的黏附，但甘露糖、葡聚糖、几丁质、伴刀豆球蛋白A或高电荷多氨基酸则无此作用。用胰凝乳蛋白酶在中性粒细胞与念珠菌孵育前预处理念珠菌，也可抑制二者接触，但胰蛋白酶或几种蛋白酶抑制剂则无此作用。对中性粒细胞进行预处理也得到类似结果，只是胰蛋白酶有抑制作用。用紫外线杀死假菌丝后，可释放出分子量<10,000的蛋白质多糖复合物，这些复合物似乎可结合到中性粒细胞表面，抑制中性粒细胞与念珠菌以及其他真菌的接触。已知作用于中性粒细胞氧化杀菌机制的试剂，包括氰化钠、叠氮化钠、过氧化氢酶、超氧化物歧化酶和单线态氧猝灭剂1,4-二氮杂双环[2.2.2]辛烷，可抑制中性粒细胞对念珠菌的损伤。慢性肉芽肿病患者的中性粒细胞根本不损伤念珠菌。然而，羟基自由基清除剂甘露醇和苯甲酸盐并无抑制作用。阳离子蛋白和乳铁蛋白在该系统中似乎也不起主要作用。低浓度溶菌酶在等渗缓冲溶液中不损伤念珠菌，但在蒸馏水中可损伤假菌丝。分离的中性粒细胞颗粒仅在添加过氧化氢和卤化物时才损伤假菌丝，且仅含髓过氧化物酶的颗粒部分才有损伤作用。这些发现表明，中性粒细胞识别念珠菌表面一种具有对胰凝乳蛋白酶敏感的蛋白质成分的分子，该分子可能在菌体死亡时从细胞表面释放出来。中性粒细胞对念珠菌的受体似乎对胰蛋白酶和胰凝乳蛋白酶敏感。中性粒细胞对念珠菌的损伤主要通过氧化机制发生，包括超氧化物和过氧化氢与髓过氧化物酶和卤化物相互作用，以及单线态氧，但似乎不涉及羟基自由基。在某些情况下，溶菌酶可能起辅助作用。
