Université de Lyon, Lyon, France.
J Appl Microbiol. 2010 Jun;108(6):2133-41. doi: 10.1111/j.1365-2672.2009.04621.x. Epub 2009 Nov 10.
The Wolbachia strain wMel can protect Drosophila melanogaster against pathogenic RNA viruses. To analyse the potential of this inhibitory effect against arboviruses vectorized by these mosquitoes, we here first transinfected the Aedes albopictus Aa23 and C6/36 cell lines with the Wolbachia strain wMel and then monitored their infection dynamics.
Wolbachia strain wMel was transferred into A. albopictus Aa23 and C6/36 cell lines using the shell vial technique. The presence of the bacterium in the transinfected cells was monitored by quantitative PCR and fluorescence in situ hybridization. Bacteria could be detected in the cytoplasm of both the Aa23 and C6/36 cell lines. However, the dynamics and stability of the bacterial infection differed depending on the initial cell background. The Aa23 cell line, which had been treated with a tetracycline antibiotic 2 years previously to eliminate its natural Wolbachia wAlbB-infecting strain, lost the introduced Wolbachia wMel strain after 12 passages postinfection. In contrast, the C6/36 cell line, which had originally been aposymbiotic, displayed a stable infection with Wolbachia wMel. The bacterial density in C6/36 was greater than that of the A. albopictus RML12 cell line from which the wMel strain had originated.
Transient or persistent transinfection of A. albopictus Aa23 and C6/36 cell lines with Wolbachia wMel strain was achieved. The results indicate the influence of the genetic background of mosquito cells in maintaining Wolbachia originating from a distant dipteral host.
The cell model built here can now be used to investigate the viral inhibitory effect of the Wolbachia wMel strain against arboviruses such as dengue and chikungunya, which are transmitted by the mosquito A. albopictus.
沃尔巴克氏体菌株 wMel 可以保护黑腹果蝇免受致病性 RNA 病毒的侵害。为了分析这种抑制作用对这些蚊子传播的虫媒病毒的潜在影响，我们首先使用壳瓶技术将沃尔巴克氏体菌株 wMel 转染到白纹伊蚊 Aa23 和 C6/36 细胞系中，然后监测它们的感染动态。
使用壳瓶技术将沃尔巴克氏体菌株 wMel 转移到白纹伊蚊 Aa23 和 C6/36 细胞系中。通过定量 PCR 和荧光原位杂交监测转染细胞中细菌的存在。细菌可以在 Aa23 和 C6/36 细胞系的细胞质中检测到。然而，细菌感染的动态和稳定性取决于初始细胞背景。两年前，为了消除其天然的沃尔巴克氏体 wAlbB 感染株，用四环素抗生素处理过的 Aa23 细胞系在感染后 12 个传代后失去了引入的沃尔巴克氏体 wMel 株。相比之下，最初为无共生体的 C6/36 细胞系显示出与沃尔巴克氏体 wMel 的稳定感染。C6/36 中的细菌密度大于其起源的白纹伊蚊 RML12 细胞系。
成功实现了白纹伊蚊 Aa23 和 C6/36 细胞系对沃尔巴克氏体 wMel 菌株的短暂或持续转染。结果表明，蚊子细胞的遗传背景对白纹伊蚊来源的沃尔巴克氏体的维持有影响。
这里建立的细胞模型现在可用于研究沃尔巴克氏体 wMel 菌株对登革热和基孔肯雅热等虫媒病毒的抑制作用，这些病毒由白纹伊蚊传播。
