Moreno-Altamirano María M B, Sánchez-García F Javier, Legorreta-Herrera Martha, Aguilar-Carmona Israel
Departamento de Inmunología, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, México, D.F., México.
Intervirology. 2007;50(3):237-9. doi: 10.1159/000100567. Epub 2007 Mar 14.
The aim of this study was to investigate whether the J774 mouse macrophage cell line could be used as an in vitro model for dengue virus infection (DENV). After 3 days, infection in J774 cells was assessed by detecting dengue virus non-structural protein 1 (NSP-1) production either by dot blot or indirect immunofluorescence assay (IFA) of saponine-permeabilized J774 cells and then confirmed by RT-PCR (171 bp product, corresponding to the DENV-2 core). Based on the presence of NSP-1 in infected but not in non-infected cells by both IFA and dot blot, as well as the amplification of a 171-bp DENV-2-specific RT-PCR product exclusively in the infected cells, the J774 cell line was found to be permissive for dengue virus infection. As far as we know, this is the first report that the J774 mouse macrophage cell line is infected with dengue virus and, thus, that it can be used as an alternative in vitro model for dengue virus infection studies. This finding could help to further elucidate the mechanisms involved in dengue virus infection and pathogenesis.
本研究的目的是调查J774小鼠巨噬细胞系是否可作为登革病毒感染(DENV)的体外模型。3天后,通过斑点印迹法或对经皂角苷通透处理的J774细胞进行间接免疫荧光分析(IFA)来检测登革病毒非结构蛋白1(NSP-1)的产生,从而评估J774细胞中的感染情况,然后通过RT-PCR(171 bp产物,对应于DENV-2核心)进行确认。基于IFA和斑点印迹法在感染细胞而非未感染细胞中检测到NSP-1,以及仅在感染细胞中扩增出171 bp的DENV-2特异性RT-PCR产物,发现J774细胞系对登革病毒感染具有易感性。据我们所知,这是关于J774小鼠巨噬细胞系感染登革病毒的首次报道,因此它可作为登革病毒感染研究的替代体外模型。这一发现有助于进一步阐明登革病毒感染和发病机制中涉及的机制。