Campos M C, Gesztesi J L, Vincentini A P, Lopes J D, Camargo Z P
Universidade Federal de São Paulo, Escola Paulista de Medicina, Disciplina de Biologia Celular, São Paulo, Brazil.
J Med Vet Mycol. 1995 Jul-Aug;33(4):223-7. doi: 10.1080/02681219580000461.
Individual exoantigens from 13 Paracoccidioides brasiliensis isolates were obtained and tested against a panel of 50 sera from patients with paracoccidioidomycosis by immunodiffusion test. The index of positivity varied from 20 to 100% according to the isolate. When these exoantigens were analysed by SDS-PAGE, eight presented high amounts of the glycoprotein gp43, two presented small amounts and in three there was no detectable gp43. The eight isolates presenting high amounts of gp43 were submitted to isoelectric focusing, blotted to nitrocellulose membranes, and revealed by monoclonal and polyclonal anti-gp43 antibodies. Four gp43 isoform profiles were obtained: profile A presenting pIs of 6.0, 6.2, 6.6 and 7.0, profile B presenting pIs of 6.4, 6.8 and 7.2, profile C presenting pI > 8.5 and profile D, presenting pIs of 5.8, 6.2 and 6.6. In each pattern observed, a major band with a distinct pI was detected. Despite this variation, our results strongly suggest an epitopic conservation among all isoforms analysed, since all of them were recognized by anti-gp43 monoclonal antibodies.
从13株巴西副球孢子菌分离物中获取了个体外抗原,并通过免疫扩散试验,用一组来自副球孢子菌病患者的50份血清对其进行检测。根据分离物的不同,阳性指数在20%至100%之间变化。当通过SDS-PAGE分析这些外抗原时,8株呈现大量糖蛋白gp43,2株呈现少量,3株未检测到gp43。对呈现大量gp43的8株分离物进行等电聚焦,转移至硝酸纤维素膜上,并用单克隆和多克隆抗gp43抗体进行显色。获得了4种gp43同工型图谱:图谱A的等电点为6.0、6.2、6.6和7.0,图谱B的等电点为6.4、6.8和7.2,图谱C的等电点>8.5,图谱D的等电点为5.8、6.2和6.6。在观察到的每种模式中,均检测到一条具有独特等电点的主要条带。尽管存在这种差异,但我们的结果强烈表明,在所分析的所有同工型中存在表位保守性,因为它们均被抗gp43单克隆抗体识别。