Figueroa J I, Hamilton A J, Allen M H, Hay R J
St John's Institute of Dermatology, Guys' Hospital, London, UK.
Trans R Soc Trop Med Hyg. 1995 Sep-Oct;89(5):566-72. doi: 10.1016/0035-9203(95)90111-6.
A novel 58 kDa antigenic determinant of the fungus Paracoccidioides brasiliensis was identified by enzyme-linked immunosorbent assay using a panel of species-specific murine monoclonal antibodies (MAbs). Western immunoblot analysis, deglycosylation studies and isoelectric focusing indicated that this 58 kDa antigen is a glycoprotein, with a pI of approximately 5.2. The molecule was purified from P. brasiliensis culture filtrate and yeast cytoplasmic antigens by membrane ultrafiltration, liquid isoelectric focusing and gel filtration; N-terminal amino acid sequence data revealed no substantial homology with known proteins. The presence of the antigen in the cytoplasm of both yeast and mycelial forms of the fungus was demonstrated when these MAbs were used as markers in immunofluorescence, immunoperoxidase and immunoalkaline phosphatase techniques to label P. brasiliensis in cryostat sections. These MAbs also recognized the cytoplasm of P. brasiliensis yeast forms in paraffin-embedded pathological specimens from human cases. A preparation of the 58 kDa component from yeast cytoplasmic antigen was reacted by Western immunoblotting with 26 different serum samples from paracoccidioidomycosis patients, and 81% of them recognized it.
通过使用一组种特异性鼠单克隆抗体（MAbs）的酶联免疫吸附测定法，鉴定出巴西副球孢子菌一种新的58 kDa抗原决定簇。蛋白质免疫印迹分析、去糖基化研究和等电聚焦表明，这种58 kDa抗原是一种糖蛋白，其等电点约为5.2。通过膜超滤、液相等电聚焦和凝胶过滤从巴西副球孢子菌培养滤液和酵母细胞质抗原中纯化该分子；N端氨基酸序列数据显示与已知蛋白质无明显同源性。当这些MAbs用作免疫荧光、免疫过氧化物酶和免疫碱性磷酸酶技术中的标记物以标记恒冷箱切片中的巴西副球孢子菌时，证明该抗原存在于该真菌酵母型和菌丝型的细胞质中。这些MAbs还识别来自人类病例的石蜡包埋病理标本中巴西副球孢子菌酵母型的细胞质。用来自副球孢子菌病患者的26种不同血清样本通过蛋白质免疫印迹法与酵母细胞质抗原的58 kDa组分制剂反应，其中81%的血清样本能识别该抗原。
