Garner R E, Rubanowice K, Sawyer R T, Hudson J A
Division of Biomedical Science, Mercer University School of Medicine, Macon, Georgia 31207.
J Leukoc Biol. 1994 Feb;55(2):161-8. doi: 10.1002/jlb.55.2.161.
Resident alveolar macrophages (AM phi) were tested for their ability to respond to Candida albicans mannan. AM phi were found to produce tumor necrosis factor alpha (TNF-alpha) in vitro in response to mannan stimulation. TNF-alpha secretion was measured using ELISA and L929B cellular cytotoxicity assays. Cytotoxicity was neutralized in parallel L929B cell cultures by the addition of rabbit anti-TNF-alpha antibody. Mannan preparations were found to be free of contaminating LPS by Limulus assay. When AM phi were cultivated for 18 h at 37 degrees C, 67 micrograms of mannan stimulated the secretion of approximately 207 U/ml of TNF-alpha. By comparison, AM phi treated with 6.7 micrograms of LPS secreted approximately 257 U/ml of TNF-alpha. Optimal TNF-alpha production occurred between 9 and 18 h after mannan stimulation. Disparate mechanisms for stimulation of TNF-alpha secretion were suggested by differential sugar blockade of LPS- and mannan-induced TNF-alpha secretion. The addition of 2% D-mannose or 2% alpha-methyl-D-mannoside to AM phi cultures blocked mannan- but not LPS-stimulated TNF-alpha secretion. Furthermore, the addition of rabbit anti-mannan antibody to mannan-coated plastic culture dishes prevented TNF-alpha secretion by the mannan-sensitive RAW 264.7 cell line. Moreover, the data suggest that C. albicans mannan stimulated AM phi to secrete TNF-alpha by an LPS-independent receptor mechanism which may also function as a mannose receptor.
检测了驻留肺泡巨噬细胞（AM phi）对白色念珠菌甘露聚糖的反应能力。发现AM phi在体外对甘露聚糖刺激有反应，能产生肿瘤坏死因子α（TNF-α）。使用酶联免疫吸附测定（ELISA）和L929B细胞毒性测定法测量TNF-α的分泌。通过添加兔抗TNF-α抗体，在平行的L929B细胞培养物中中和细胞毒性。通过鲎试剂测定法发现甘露聚糖制剂不含脂多糖污染。当AM phi在37℃培养18小时时，67微克甘露聚糖刺激分泌约207 U/ml的TNF-α。相比之下，用6.7微克脂多糖处理的AM phi分泌约257 U/ml的TNF-α。TNF-α的最佳产生发生在甘露聚糖刺激后9至18小时之间。脂多糖和甘露聚糖诱导的TNF-α分泌的不同糖阻断提示了刺激TNF-α分泌的不同机制。向AM phi培养物中添加2% D-甘露糖或2% α-甲基-D-甘露糖苷可阻断甘露聚糖而非脂多糖刺激的TNF-α分泌。此外，向包被有甘露聚糖的塑料培养皿中添加兔抗甘露聚糖抗体可阻止甘露聚糖敏感的RAW 264.7细胞系分泌TNF-α。此外，数据表明白色念珠菌甘露聚糖通过一种不依赖脂多糖的受体机制刺激AM phi分泌TNF-α，该机制可能也作为一种甘露糖受体发挥作用。
