Loizel Christophe, Blanchard Philippe, Grasland Béatrice, Dory Daniel, Oger Aurélie, Nignol Anne-Cécile, Cariolet Roland, Jestin André
The Unit of Viral Genetics and Biosafety, French Agency for Food Safety, Ploufragan, France.
Int J Exp Pathol. 2005 Feb;86(1):33-43. doi: 10.1111/j.0959-9673.2005.00409.x.
Post-weaning multisystemic wasting syndrome (PMWS) is a complex disease syndrome in swine, affecting nursery and fattening pigs. Although ongoing evidence suggests that porcine circovirus type-2 (PCV2) is the causal agent of PMWS, the host immune system appears to have a crucial role in the PMWS pathogenesis of PCV2-affected pigs. Owing to difficulties in producing a biologically pure form of PCV2 devoid of the other viral agents commonly present in swine tissues, we decided to use a tandem-cloned PCV2 DNA providing highly pure grade reagent in order to monitor the virulence of PCV2 alone or with an immunostimulating co-factor, granulocyte-macrophage colony-stimulating factor (GM-CSF). A single intramuscular injection of tandem-cloned PCV2 DNA into 5-week-old piglets produced plasmid to viral genome progeny and infectious particles as early as 8 days post-injection in all the organs tested (the lung, the tonsil and the inguinal, mesenteric, bronchial and upper-right axial lymph nodes). The initial plasmid load was not detected with the help of primers designed to specifically detect the acceptor plasmid, thus confirming the replication of the viral genome. Despite the presence of a high level of PCV2 genome copies in the lymphoid organs--the tonsil and the lung--and the presence of infectious particles, no detectable clinical manifestations or pathological lesions were observed in the transfected pigs over the period of observation, regardless of whether they had been co-injected with plasmid containing GM-CSF DNA or had received plasmid containing PCV2 DNA alone. GM-CSF encoding DNA injection had no significant effect on viral replication or on the production of viral particles and appearance of the disease.
断奶后多系统消耗综合征(PMWS)是猪的一种复杂疾病综合征,影响保育猪和育肥猪。尽管现有证据表明2型猪圆环病毒(PCV2)是PMWS的病原体,但宿主免疫系统似乎在受PCV2感染猪的PMWS发病机制中起关键作用。由于难以生产不含猪组织中常见其他病毒因子的生物纯形式的PCV2,我们决定使用串联克隆的PCV2 DNA来提供高纯度试剂,以便单独监测PCV2或与免疫刺激辅因子粒细胞巨噬细胞集落刺激因子(GM-CSF)一起的毒力。对5周龄仔猪单次肌内注射串联克隆的PCV2 DNA,早在注射后8天,在所有测试器官(肺、扁桃体以及腹股沟、肠系膜、支气管和右上腹淋巴结)中就产生了从质粒到病毒基因组后代和感染性颗粒。用专门设计用于检测受体质粒的引物未检测到初始质粒负荷,从而证实了病毒基因组的复制。尽管在淋巴器官(扁桃体和肺)中存在高水平的PCV2基因组拷贝以及存在感染性颗粒,但在观察期内,无论转染猪是与含GM-CSF DNA的质粒共注射还是仅接受含PCV2 DNA的质粒,均未观察到可检测到的临床表现或病理损伤。注射编码GM-CSF的DNA对病毒复制、病毒颗粒产生和疾病出现没有显著影响。