LaBoissière S, Trudel M, Simard C
Centre de Recherche en Virologie, Institut Armand-Frappier, Université du Québec, Laval, Canada.
Virus Res. 1996 Feb;40(2):191-8. doi: 10.1016/0168-1702(95)01273-7.
We previously reported the characterization of the gene encoding the bovine herpesvirus type 1 (BHV-1) major tegument protein VP8. With the aim of defining the immunological properties of this protein, we constructed a recombinant vaccinia virus (VV-VP8) in which expression of the VP8 gene was regulated by the P7.5 early/late promoter. Since the sequence of the VP8 gene contained a TTTTTNT motif known to serve as a transcription termination signal of vaccinia virus genes of the early class, a second recombinant (VV-VP8-Mut) in which this signal was modified by site-directed mutagenesis was created. Characterization of the recombinant viruses revealed that truncated VP8 mRNA and protein (69 kDa) were synthesized in VV-VP8 infected cells, whereas cells infected with VV-VP8-Mut produced a protein which was undistinguishable from that of the BHV-1 encoded protein (92-94 kDa). Immunization of BALB/c mice (H-2d) with VV-VP8-Mut induced a low VP8-specific antibody response whereas no specific response was induced in VV-VP8 inoculated mice. The low humoral response elicited was similar in C57BL/6 (H-2b) and C3H (H-2k) mice. Furthermore, immunization of mice with VV-VP8-Mut did not induce a BHV-1-specific lymphoproliferation in the three mice strains examined. Our results contrast with a recent study showing that immunization of calves with purified VP8 stimulated both T cell proliferation and antibody production.
我们之前报道了编码牛疱疹病毒1型(BHV-1)主要被膜蛋白VP8的基因的特征。为了确定该蛋白的免疫学特性,我们构建了一种重组痘苗病毒(VV-VP8),其中VP8基因的表达由P7.5早期/晚期启动子调控。由于VP8基因的序列包含一个已知作为早期类痘苗病毒基因转录终止信号的TTTTTNT基序,因此构建了第二个重组体(VV-VP8-Mut),其中通过定点诱变对该信号进行了修饰。重组病毒的特性表明,在VV-VP8感染的细胞中合成了截短的VP8 mRNA和蛋白(69 kDa),而感染VV-VP8-Mut的细胞产生的一种蛋白与BHV-1编码的蛋白(92-94 kDa)无法区分。用VV-VP8-Mut免疫BALB/c小鼠(H-2d)诱导了低水平的VP8特异性抗体反应,而在接种VV-VP8的小鼠中未诱导出特异性反应。在C57BL/6(H-2b)和C3H(H-2k)小鼠中引发的低体液反应相似。此外,用VV-VP8-Mut免疫小鼠在检测的三种小鼠品系中均未诱导出BHV-1特异性淋巴细胞增殖。我们的结果与最近一项研究形成对比,该研究表明用纯化的VP8免疫小牛可刺激T细胞增殖和抗体产生。