Barderas M G, Rodríguez F, Gómez-Puertas P, Avilés M, Beitia F, Alonso C, Escribano J M
Departamento de Biotecnología, INIA, Madrid, Spain.
Arch Virol. 2001;146(9):1681-91. doi: 10.1007/s007050170056.
A chimera of the two immunodominant African swine fever (ASF) virus proteins p54 and p30 was constructed by insertion of the gene CP204L into a Not I restriction site of E183L gene. The resulting chimeric protein p54/30, expressed by a recombinant baculovirus in insect cells and in Trichoplusia ni larvae, retained antigenic determinants present in both proteins and reacted in Western blot with a collection of sera from inapparent ASF virus carrier pigs. Remarkably, pigs immunized with the chimeric protein developed neutralizing antibodies and survived the challenge with a virulent African swine fever virus, presenting a reduction of about two logs in maximum viremia titers with respect to control pigs. In conclusion, this study revealed that the constructed chimeric protein may have utility as a serological diagnostic reagent and for further immunological studies that may provide new insights on mechanisms of protective immunity to ASFV.
通过将CP204L基因插入E183L基因的Not I限制性酶切位点,构建了两种免疫显性非洲猪瘟(ASF)病毒蛋白p54和p30的嵌合体。由重组杆状病毒在昆虫细胞和粉纹夜蛾幼虫中表达产生的嵌合蛋白p54/30,保留了两种蛋白中存在的抗原决定簇,并在蛋白质印迹法中与来自隐性ASF病毒携带猪的血清样本发生反应。值得注意的是,用该嵌合蛋白免疫的猪产生了中和抗体,并在强毒非洲猪瘟病毒攻击下存活下来,与对照猪相比,最大病毒血症滴度降低了约两个对数。总之,本研究表明,构建的嵌合蛋白可作为血清学诊断试剂,并用于进一步的免疫学研究,这些研究可能为ASFV保护性免疫机制提供新的见解。