EXPRESSION OF BOVINE HERPESVIRUS-1 GLYCOPROTEIN GIV BY RECOMBINANT BACULOVIRUS AND ANALYSIS OF ITS IMMUNOGENIC PROPERTIES
EXPRESSION OF BOVINE HERPESVIRUS-1 GLYCOPROTEIN GIV BY RECOMBINANT BACULOVIRUS AND ANALYSIS OF ITS IMMUNOGENIC PROPERTIES
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DOI：
10.1128/jvi.65.1.263-271.1991
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发表时间：
1991-01-01
影响因子：
5.4
通讯作者：
BABIUK, LA
中科院分区：
文献类型：
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作者：
LITTELVANDENHURK, SV;PARKER, MD;BABIUK, LA
编码牛疱疹病毒1型gIV糖蛋白的基因已被插入到加利福尼亚自签名虫杆状病毒基因组中，以代替加利福尼亚自签名虫杆状病毒多面蛋白基因的编码区。通过与giv特异性单克隆抗体的反应性鉴定了重组蛋白，并在Spodoptera frugiperda （SF9）细胞中高水平表达（约85微克/ 2.5 × 10(6)个细胞）。重组糖蛋白的表观分子量为63 kDa，表明其糖基化不完全。然而，它被转运到感染的SF9细胞表面并在细胞表面表达。此外，与gIV特异性多克隆和单克隆抗体的反应性表明，重组gIV的大多数表位在功能上没有改变。用粗的、部分纯化的或纯形式的重组gIV免疫牛，可诱导BHV-1的中和抗体，这些抗体与真gIV反应。然而，中和抗体滴度低于等量亲和纯化的真gIV，这似乎主要是由于对gIV的一个中和抗原结构域（指定结构域i）的识别降低。本文讨论了重组gIV糖蛋白作为牛疱疹病毒1型感染疫苗的潜在用途。
The gene encoding the gIV glycoprotein of bovine herpesvirus 1 has been inserted into the genome of Autographa californica baculovirus in lieu of the coding region of the A. californica baculovirus polyhedrin gene. Recombinant protein was identified by its reactivity with gIV-specific monoclonal antibodies and expressed at high levels (about 85 micrograms per 2.5 x 10(6) cells) in Spodoptera frugiperda (SF9) cells. The recombinant glycoprotein had an apparent molecular mass of 63 kDa, indicating that it was incompletely glycosylated. However, it was transported to and expressed on the cell surface of infected SF9 cells. Furthermore, reactivity with polyclonal and monoclonal antibodies specific for gIV suggested that most epitopes were functionally unaltered on the recombinant gIV. Immunization of cattle with recombinant gIV in crude, partially purified, or pure form resulted in the induction of neutralizing antibodies to BHV-1, which were reactive with authentic gIV. However, the neutralizing antibody titers were lower than those elicited by an equivalent amount of affinity-purified authentic gIV, which appeared to be mainly due to reduced recognition of one of the neutralizing antigenic domains of gIV, designated domain I. The potential use of this recombinant gIV glycoprotein as a vaccine to bovine herpesvirus 1 infection in cattle is discussed.