Ellsworth J H, Reiss E, Bradley R L, Chmel H, Armstrong D
J Clin Microbiol. 1977 Jan;5(1):91-9. doi: 10.1128/jcm.5.1.91-99.1977.
Yeast-form Candida albicans cells were disrupted for 1.5 min in a Braun homogenizer and centrifuged at 100,000 X g. The supernatant was concentrated by ammonium sulfate precipitation and then dialyzed. The resulting material (650 mg), containing 81.2% protein and 11.5% carbohydrate, was subjected to affinity chromatography on concanavalin A (Con A) linked to agarose. A protein fraction was eluted from the column with buffer, and a fraction containing mannan was eluted with 0.2 M alpha-methyl mannoside. The candidal soluble proteins had 19 components which were resolvable by polyacrylamide gel electrophoresis. The material with affinity for Con A contained mannan and 17% complexed protein. Antigenic differences between the soluble proteins and the mannan-protein complex were shown by lines of intersection in immunodiffusion. The soluble proteins devoid of mannan reacted in immunoelectrophoresis with sera from infected rabbits and patients with chronic candidiasis. These same sera also reacted with a mannan-protein complex eluted from the Con A column with alpha-methyl mannoside. The comparative ability of candidal proteins and cell wall-derived mannan to elicit skin test reactions in guinea pigs sensitized by infection or with formaldehyde-killed yeast was studied. Candidal proteins at a 10-mug dose elicited positive reactions at 6 and 21 days after sensitization. The reactions persisted for 48 h and showed minimal tendency to an arthus response, which was marked when mannan-containing antigens were used. The antigenicity of cell wall-derived mannans and candidal soluble proteins devoid of mannan was compared in immunodiffusion tests of sera from 39 patients with neoplastic disease. Of these patients with documented candidiasis, 13 of 20 reacted to one or more mannan antigens, and 3 of 20 reacted to candidal soluble proteins. In contrast, of those patients who were uninfected or had superficial Candida spp. infections, 5 of 19 reacted to candidal soluble proteins, and 16 of 19 reacted to one or more mannan antigens.
白色念珠菌酵母样细胞在Braun匀浆器中匀浆1.5分钟，然后在100,000×g下离心。上清液通过硫酸铵沉淀浓缩，然后透析。得到的物质（650毫克），含有81.2%的蛋白质和11.5%的碳水化合物，进行与琼脂糖偶联的伴刀豆球蛋白A（Con A）亲和层析。用缓冲液从柱上洗脱一个蛋白质组分，用0.2Mα-甲基甘露糖苷洗脱一个含甘露聚糖的组分。念珠菌可溶性蛋白有19个组分，可通过聚丙烯酰胺凝胶电泳分离。对Con A有亲和力的物质含有甘露聚糖和17%的复合蛋白。免疫扩散中的交叉线显示了可溶性蛋白和甘露聚糖-蛋白复合物之间的抗原差异。不含甘露聚糖的可溶性蛋白在免疫电泳中与感染兔和慢性念珠菌病患者的血清发生反应。这些相同的血清也与用α-甲基甘露糖苷从Con A柱上洗脱的甘露聚糖-蛋白复合物发生反应。研究了念珠菌蛋白和细胞壁来源的甘露聚糖在感染致敏或甲醛灭活酵母致敏的豚鼠中引发皮肤试验反应的比较能力。10微克剂量的念珠菌蛋白在致敏后6天和21天引发阳性反应。反应持续48小时，显示出最小的阿瑟斯反应倾向，而使用含甘露聚糖的抗原时这种倾向很明显。在对39例肿瘤疾病患者血清的免疫扩散试验中，比较了细胞壁来源的甘露聚糖和不含甘露聚糖的念珠菌可溶性蛋白的抗原性。在这些有记录的念珠菌病患者中，20例中有13例对一种或多种甘露聚糖抗原发生反应，20例中有3例对念珠菌可溶性蛋白发生反应。相比之下，在那些未感染或有浅表念珠菌属感染的患者中，19例中有5例对念珠菌可溶性蛋白发生反应，19例中有16例对一种或多种甘露聚糖抗原发生反应。
