Gauss-Müller V, Zhou M Q, von der Helm K, Deinhardt F
Institute for Medical Microbiology, Medical University of Lübeck, Federal Republic of Germany.
J Med Virol. 1990 Aug;31(4):277-83. doi: 10.1002/jmv.1890310407.
Six overlapping genomic regions of capsid proteins VP1 and VP3 of hepatitis A virus (HAV) inserted into the expression vectors pBD or pUR respectively expressed beta-galactosidase-HAV fusion proteins. The recombinant proteins were poorly soluble so they were difficult to detect by human anti-HAV sera in radioimmunoassay, but the fusion proteins dissolved in sodium dodecyl sulfate reacted with human and rabbit anti-HAV-positive sera in immunoblots. Antisera against VP1 and VP3 recombinant proteins reacted with the respective structural proteins of HAV in immunoblots. Two recombinant proteins, one including the first 120 amino acids of the N-terminus of VP1 and the other containing all of VP1 except for the first 60 N-terminal amino acids, induced a transient neutralizing antibody response in rabbits. Antisera directed against other regions of VP1 and VP3 neither neutralized viral infectivity nor recognized native virus in a competitive radioimmunoassay. However, when immunized animals were challenged with a sub-immunogenic dose of HAV, all animals responded with stable virus-neutralizing antibodies.
甲型肝炎病毒（HAV）衣壳蛋白VP1和VP3的六个重叠基因组区域分别插入表达载体pBD或pUR中，表达β-半乳糖苷酶-HAV融合蛋白。重组蛋白溶解性差，因此在放射免疫分析中难以被人抗HAV血清检测到，但溶解在十二烷基硫酸钠中的融合蛋白在免疫印迹中与人及兔抗HAV阳性血清发生反应。抗VP1和VP3重组蛋白的抗血清在免疫印迹中与HAV各自的结构蛋白发生反应。两种重组蛋白，一种包含VP1 N端的前120个氨基酸，另一种包含除N端前60个氨基酸外的所有VP1，在兔中诱导了短暂的中和抗体反应。针对VP1和VP3其他区域的抗血清在竞争性放射免疫分析中既不能中和病毒感染性，也不能识别天然病毒。然而，当用亚免疫原剂量的HAV攻击免疫动物时，所有动物都产生了稳定的病毒中和抗体。
