Leijh P C, van den Barselaar M T, van Furth R
Infect Immun. 1977 Aug;17(2):313-8. doi: 10.1128/iai.17.2.313-318.1977.
The study of the phagocytosis and intracellular killing of Candida albicans by granulocytes and monocytes has been hampered by the budding and pseudomycelium formation of this yeast during a relatively short incubation period at 37 degrees C and by the similar density of candida cells and phagocytes, which makes differential centrifugation impossible. In the present study, C. albicans was used after 5 days of preculture at 30 degrees C, after which the number of candida cells remained constant during incubation at 37 degrees C for 90 min. On this basis, phagocytosis and intracellular killing were limited to a period of 60 min. Phagocytosis of C. albicans by granulocytes and monocytes was measured with a hemocytometer, the number of extracellular candida being a measure of the ingestion of these microorganisms. After 60 min, 96% of the candida cells were ingested by normal human granulocytes and monocytes. This process was dependent on the opsonin concentration and temperature and was inhibited by mono-iodoacetic acid. Heat-inactivated serum was less active than fresh serum, reflecting the role of complement factors with respect to opsonization. Intracellular killing was measured by a microbiological assay. After 60 min of incubation of phagocytes together with C. albicans and serum, human granulocytes and monocytes killed 58 and 50% of the ingested candida, respectively. This process was inhibited by phenylbutazone. Phagocytes from patients with chronic granulomatous disease showed impaired intracellular killing.
粒细胞和单核细胞对白色念珠菌的吞噬作用及细胞内杀伤作用的研究受到了阻碍，这是因为在37℃相对较短的孵育期内，这种酵母菌会出芽并形成假菌丝，而且念珠菌细胞和吞噬细胞的密度相似，这使得差速离心法无法进行。在本研究中，白色念珠菌在30℃预培养5天后使用，此后在37℃孵育90分钟期间念珠菌细胞数量保持恒定。在此基础上，吞噬作用和细胞内杀伤作用被限制在60分钟内。用血细胞计数器测量粒细胞和单核细胞对白色念珠菌的吞噬作用，细胞外念珠菌的数量可作为这些微生物摄取量的指标。60分钟后，96%的念珠菌细胞被正常人的粒细胞和单核细胞摄取。这个过程取决于调理素浓度和温度，并受到一碘乙酸的抑制。热灭活血清的活性低于新鲜血清，这反映了补体因子在调理作用方面的作用。通过微生物学检测来测量细胞内杀伤作用。吞噬细胞与白色念珠菌和血清一起孵育60分钟后，人类粒细胞和单核细胞分别杀死了58%和50%被摄取的念珠菌。这个过程受到保泰松的抑制。慢性肉芽肿病患者的吞噬细胞显示出细胞内杀伤功能受损。
