A method for the generation of ectromelia virus (ECTV) recombinants: in vivo analysis of ECTV vCD30 deletion mutants.
A method for the generation of ectromelia virus (ECTV) recombinants: in vivo analysis of ECTV vCD30 deletion mutants.
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DOI：
10.1371/journal.pone.0005175
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发表时间：
2009
期刊：
影响因子：
3.7
通讯作者：
Alcami A
中科院分区：
文献类型：
--
作者：
Alejo A;Saraiva M;Ruiz-Argüello MB;Viejo-Borbolla A;de Marco MF;Salguero FJ;Alcami A
鼠痘是一种与人类天花相似的小鼠致死性疾病，鼠痘病毒（ECTV）是其病原体。鼠痘进展涉及在感染的初始部位（通常是皮肤）复制，随后迅速扩散到次级复制器官（脾和肝），最后扩散到皮肤，在皮肤处出现与这种和其他正痘病毒诱导的疾病相关的典型皮疹。病死率由基因决定，在易感小鼠品系中高达100%。与其他痘病毒一样，ECTV编码许多具有免疫调节潜力的蛋白质，其在鼠痘进展中的作用在很大程度上尚未描述。其中有细胞肿瘤坏死因子受体超家族成员CD 30的分泌同源物，已提出其在体内调节Th 1免疫应答。为了评估病毒CD 30（vCD 30）在感染宿主中对病毒发病机制的贡献，我们采用了一种新的瞬时显性方法来选择重组ECTV。使用这种方法，我们已经产生了ECTV vCD 30缺失突变体，其相应的回复突变对照病毒以及编码鼠CD 30胞外结构域的病毒。这些病毒在其基因组中不含外源标记DNA序列，与迄今报道的其他ECTV相反。我们表明，vCD 30表达为分泌的二硫键连接的三聚体，并且vCD 30的缺乏不会损害鼠痘诱导的体内死亡率。用分泌形式的小鼠CD 30替代vCD 30导致ECTV的有限衰减。产生的重组病毒可用于研究细胞CD 30-CD 30 L相互作用在免疫应答发展中的作用。所开发的方法可能是有用的构建ECTV突变体的其他基因的研究。
Ectromelia virus (ECTV) is the causative agent of mousepox, a lethal disease of mice with similarities to human smallpox. Mousepox progression involves replication at the initial site of infection, usually the skin, followed by a rapid spread to the secondary replicative organs, spleen and liver, and finally a dissemination to the skin, where the typical rash associated with this and other orthopoxviral induced diseases appears. Case fatality rate is genetically determined and reaches up to 100% in susceptible mice strains. Like other poxviruses, ECTV encodes a number of proteins with immunomodulatory potential, whose role in mousepox progression remains largely undescribed. Amongst these is a secreted homologue of the cellular tumour necrosis factor receptor superfamily member CD30 which has been proposed to modulate a Th1 immune response in vivo. To evaluate the contribution of viral CD30 (vCD30) to virus pathogenesis in the infected host, we have adapted a novel transient dominant method for the selection of recombinant ECTVs. Using this method, we have generated an ECTV vCD30 deletion mutant, its corresponding revertant control virus as well as a virus encoding the extracellular domain of murine CD30. These viruses contain no exogenous marker DNA sequences in their genomes, as opposed to other ECTVs reported up to date. We show that the vCD30 is expressed as a secreted disulfide linked trimer and that the absence of vCD30 does not impair mousepox induced fatality in vivo. Replacement of vCD30 by a secreted version of mouse CD30 caused limited attenuation of ECTV. The recombinant viruses generated may be of use in the study of the role of the cellular CD30-CD30L interaction in the development of the immune response. The method developed might be useful for the construction of ECTV mutants for the study of additional genes.
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影响因子：
3.8
作者：
Bartlett, NW;Dumoutier, L;Smith, GL
通讯作者：
Smith, GL
DOI：
10.1073/pnas.0402949101
发表时间：
2004-06-15
影响因子：
11.1
作者：
Chaudhri, G;Panchanathan, V;Karupiah, G
通讯作者：
Karupiah, G
影响因子：
3.7
作者：
CHEN, W;DRILLIEN, R;BULLER, RML
通讯作者：
BULLER, RML
影响因子：
64.5
作者：
ALCAMI, A;SMITH, GL
通讯作者：
SMITH, GL
DOI：
10.1073/pnas.0737244100
发表时间：
2003-04-15
影响因子：
11.1
作者：
Brunetti, CR;Paulose-Murphy, M;McFadden, G
通讯作者：
McFadden, G