Kouguchi Hirokazu, Suzuki Tomohiro, Yamano Kimiaki, Honma Hiroshi, Sawada Yukiharu
Hokkaido Institute of Public Health, Kita-Ku, Sapporo 060-0819, Japan.
Protein J. 2005 Jan;24(1):57-64. doi: 10.1007/s10930-004-0606-9.
Using four clones isolated from Echinococcus multilocularis cDNA library with alveolar echinococcosis (AE) patient sera, various antigens were expressed as ThioHis tag-fused protein. Recombinant EmII/3 antigen was produced as the five fragments divided into the N-terminal (#5 and #5s), the central (#6 and #6s) and the C-terminal domain (#7). Immunoblot analysis revealed that the #7 showed significant reactivity whereas those of #5 and #5s were relatively low. The #6 and #6s also showed lower reactivity than that of #7, although the two minor bands of #6 reacted with every serum. These results suggested that an immunodominant region of EmII/3 locate within the C-terminal one third. The #8s recombinant antigen, Ser23-Glu176 of actin filament fragmenting protein (AFFP), apparently reacted with the AE patient sera, while the #1 antigen synthesized as a full-length antigen BI did not show such high reactivity. Thus, #7 and #8s antigens showed significant potential for use in immunodetection of AE. In addition, the specific antibodies against #7 and #8s reacted with specific antigens in crude extract of E. multilocularis cyst, indicating that these antigens retained antigenicity common to native EmII/3 and AFFP, respectively.
利用从多房棘球绦虫cDNA文库中分离出的四个克隆与泡型包虫病(AE)患者血清,将各种抗原表达为硫氧还蛋白组氨酸标签融合蛋白。重组EmII/3抗原作为五个片段产生,分为N端(#5和#5s)、中央(#6和#6s)和C端结构域(#7)。免疫印迹分析显示,#7表现出显著的反应性,而#5和#5s的反应性相对较低。#6和#6s的反应性也低于#7,尽管#6的两条小带与每份血清都有反应。这些结果表明,EmII/3的免疫显性区域位于C端的三分之一处。#8s重组抗原,肌动蛋白丝断裂蛋白(AFFP)的Ser23-Glu176,明显与AE患者血清发生反应,而作为全长抗原BI合成的#1抗原未表现出如此高的反应性。因此,#7和#8s抗原在AE免疫检测中显示出显著的应用潜力。此外,针对#7和#8s的特异性抗体与多房棘球绦虫囊肿粗提物中的特异性抗原发生反应,表明这些抗原分别保留了天然EmII/3和AFFP共有的抗原性。