Zielonka Anja, Gedvilaite Alma, Ulrich Rainer, Lüschow Dörte, Sasnauskas Kestutis, Müller Hermann, Johne Reimar
Institute for Virology, Faculty of Veterinary Medicine, University of Leipzig, An den Tierkliniken 29, D-04103 Leipzig, Germany.
Virus Res. 2006 Sep;120(1-2):128-37. doi: 10.1016/j.virusres.2006.02.010. Epub 2006 Jun 15.
Goose hemorrhagic polyomavirus (GHPV) is the causative agent of hemorrhagic nephritis and enteritis of geese (HNEG), a fatal disease of young geese with high mortality rates. GHPV cannot be efficiently propagated in tissue culture. To provide antigens for diagnostic tests and vaccines, its major structural protein VP1 was recombinantly expressed in Sf9 insect cells and in the yeast Saccharomyces cerevisiae. As demonstrated by density gradient centrifugation and electron microscopy, GHPV-VP1 expressed in insect cells formed virus-like particles (VLPs) with a diameter of 45 nm indistinguishable from infectious polyomavirus particles. However, efficiency of VLP formation was low as compared to the monkey polyomavirus SV-40-VP1. In yeast cells, GHPV-VP1 alone formed smaller VLPs, 20 nm in diameter. Remarkably, co-expression of GHPV-VP2 resulted in VLPs with a diameter of 45 nm. All three types of GHPV-VLPs were shown to hemagglutinate chicken erythrocytes. ELISA and hemagglutination inhibition tests using the VLPs as antigen detected GHPV-specific antibodies in up to 85.7% of sera derived from flocks with HNEG but in none of the sera of a clinically healthy flock. However, GHPV-specific antibodies were also detected in sera from two other flocks without HNEG indicating a broad distribution of GHPV due to subclinical or unrecognised infections.
鹅出血性多瘤病毒(GHPV)是鹅出血性肾炎和肠炎(HNEG)的病原体,这是一种幼鹅的致命疾病,死亡率很高。GHPV无法在组织培养中有效增殖。为了提供用于诊断测试和疫苗的抗原,其主要结构蛋白VP1在Sf9昆虫细胞和酿酒酵母中进行了重组表达。通过密度梯度离心和电子显微镜证明,在昆虫细胞中表达的GHPV-VP1形成了直径为45nm的病毒样颗粒(VLP),与感染性多瘤病毒颗粒无法区分。然而,与猴多瘤病毒SV-40-VP1相比,VLP形成效率较低。在酵母细胞中,单独的GHPV-VP1形成直径为20nm的较小VLP。值得注意的是,GHPV-VP2的共表达导致形成直径为45nm的VLP。所有三种类型的GHPV-VLP均显示能凝集鸡红细胞。使用VLP作为抗原的ELISA和血凝抑制试验在高达85.7%的来自患有HNEG的鸡群的血清中检测到GHPV特异性抗体,但在临床健康鸡群的血清中均未检测到。然而,在另外两个没有HNEG的鸡群的血清中也检测到了GHPV特异性抗体,这表明由于亚临床或未被识别的感染,GHPV分布广泛。