用表达透明带C蛋白的重组痘苗病毒对欧洲赤狐进行实验性接种。

Experimental inoculation of European red foxes with recombinant vaccinia virus expressing zona pellucida C proteins.

作者信息

Reubel Gerhard H, Beaton Sandra, Venables Daryl, Pekin Jenny, Wright John, French Nigel, Hardy Christopher M

机构信息

Pest Animal Control Cooperative Research Centre, CSIRO Sustainable Ecosystems, GPO Box 284, Canberra, ACT 2601, Australia.

出版信息

Vaccine. 2005 Aug 15;23(35):4417-26. doi: 10.1016/j.vaccine.2005.04.016.

DOI:10.1016/j.vaccine.2005.04.016

PMID:15936853

Abstract

The antifertility potential of zona pellucida proteins was tested in European red foxes by immunizing females with recombinant vaccinia viruses that express zona pellucida subunit C proteins. The fox zona pellucida C (fZPC) protein was newly identified and isolated as a cDNA clone from fox ovary RNA. Eighteen European foxes were inoculated with the recombinant vaccinia viruses or with wildtype vaccinia virus (wtVV) and their clinical, virological and immune responses evaluated. Following intradermal inoculation with wtVV or recombinant vaccinia virus expressing fox zona pellucida C (rVV-fZPC), or after peroral administration with recombinant vaccinia virus expressing the porcine zona pellucida C protein (rVV-pZPC) clinical signs of disease were not observed. Five out of six foxes developed antibodies to wtVV proteins. However, none of 12 foxes (six inoculated intradermally with rVV-fZPC, six perorally with rVV-pZPC) reacted in immunoblots with the transgenic fZPC or pZPC, respectively. Infectious wtVV, rVV-fZPC or rVV-pZPC was not isolated from mucosal secretions of any of the foxes whereas viral DNA was present in oral swabs of 3/18 foxes as determined by PCR. Hematological parameters remained mostly unchanged. Histopathological changes were not observed in the ovaries of rVV-fZPC or wtVV inoculated foxes. The data indicate that inoculation of foxes with cell culture infectious wtVV, rVV-fZPC or rVV-pZPC did not result in production of infectious progeny virus in vivo. For this reason transgene expression may have been insufficient to induce adequate immune responses against the transgenic proteins.

摘要

通过用表达透明带亚基C蛋白的重组痘苗病毒免疫雌性欧洲赤狐，测试了透明带蛋白的抗生育潜力。从狐狸卵巢RNA中首次鉴定并分离出狐狸透明带C（fZPC）蛋白作为cDNA克隆。给18只欧洲赤狐接种重组痘苗病毒或野生型痘苗病毒（wtVV），并评估它们的临床、病毒学和免疫反应。用wtVV或表达狐狸透明带C的重组痘苗病毒（rVV-fZPC）进行皮内接种后，或用表达猪透明带C蛋白的重组痘苗病毒（rVV-pZPC）进行口服给药后，均未观察到疾病的临床症状。6只狐狸中有5只产生了针对wtVV蛋白的抗体。然而，12只狐狸（6只皮内接种rVV-fZPC，6只口服rVV-pZPC）中没有一只在免疫印迹中分别与转基因fZPC或pZPC发生反应。在任何一只狐狸的粘膜分泌物中均未分离到具有感染性的wtVV、rVV-fZPC或rVV-pZPC，而通过PCR测定，在18只狐狸中有3只的口腔拭子中存在病毒DNA。血液学参数大多保持不变。在接种rVV-fZPC或wtVV的狐狸卵巢中未观察到组织病理学变化。数据表明，用细胞培养感染性wtVV、rVV-fZPC或rVV-pZPC接种狐狸不会在体内产生感染性子代病毒。因此，转基因表达可能不足以诱导针对转基因蛋白的充分免疫反应。