Guerrero Nidya Alexandra Segura, Bello Felio Jesús
Universidad Pedagógica y Tecnológica de Colombia, Faculty of Science, Laboratory of Medical and Forensic Entomology, Tunja, Colombia.
Universidad de La Salle, Faculty of Agricultural and Livestock Sciences, Program of Veterinary Medicine, Bogotá, Colombia.
Rev Soc Bras Med Trop. 2019 Apr 25;52:e20180511. doi: 10.1590/0037-8682-0511-2018.
Insect cell cultures play an essential role in understanding arboviral replication. However, the replicative efficiency of some of these viruses such as dengue (DENV), yellow fever (YFV), and chikungunya (CHIKV) in a new cellular substrate (Lulo) and in the other two recognized cell lines has not been comparatively assessed.
Vero, C6/36, and Lulo cell lines were infected with DENV, YFV, and CHIKV. The viral progeny was quantified through plaque assays and quantitative reverse transcription-polymerase chain reaction, while for DENV2, the findings were confirmed by immunofluorescence antibody assay.
The higher DENV2 titer (from multiplicity of infection 0.001) was obtained on day four post-infection in C6/36 and on day six in Vero cells, while the Lulo cell line was almost impossible to infect under the same conditions. However, C6/36 showed the highest values of viral RNA production compared to Vero cells, while the quantification of the viral RNA in Lulo cells showed high levels of viral genomes, which had no correlation to the infectious viral particles.
C6/36 was the most efficient cell line in the alpha and flavivirus production, followed by Vero cells. Thus, Lulo cells may be a useful substrate to study the mechanisms by which cells evade viral replication.
昆虫细胞培养在理解虫媒病毒复制方面发挥着重要作用。然而,登革热病毒(DENV)、黄热病毒(YFV)和基孔肯雅病毒(CHIKV)等部分病毒在一种新的细胞底物(卢洛细胞)以及另外两种公认的细胞系中的复制效率尚未得到比较评估。
用DENV、YFV和CHIKV感染Vero细胞、C6/36细胞和卢洛细胞系。通过蚀斑试验和定量逆转录 - 聚合酶链反应对病毒子代进行定量,而对于DENV2,通过免疫荧光抗体试验对结果进行确认。
感染后第4天在C6/36细胞以及第6天在Vero细胞中获得了更高的DENV2滴度(感染复数为0.001时),而在相同条件下卢洛细胞系几乎无法被感染。然而,与Vero细胞相比,C6/36细胞显示出最高的病毒RNA产生值,而卢洛细胞中病毒RNA的定量显示病毒基因组水平较高,这与感染性病毒颗粒无关。
C6/36是生产甲型病毒和黄病毒最有效的细胞系,其次是Vero细胞。因此,卢洛细胞可能是研究细胞逃避病毒复制机制的有用底物。