Comparative assessment of the replication efficiency of dengue, yellow fever, and chikungunya arboviruses in some insect and mammalian cell lines

Abstract INTRODUCTION: 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. METHODS: 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. RESULTS: 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. CONCLUSIONS: 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.

Natural invertebrate hosts of iridoviruses (Iridoviridae) / Hospederos naturales de los iridovirus de invertebrados

Invertebrate iridescent viruses (IIVs) are icosahedral DNA viruses that infect invertebrates, mainly insects and terrestrial isopods, in damp and aquatic habitats. Exhaustive searches of databases resulted in the identification of 79 articles reporting 108 invertebrate species naturally infected by confirmed or putative iridoviruses. Of these, 103 (95%) were arthropods and the remainder were molluscs, an annelid worm and a nematode. Nine species were from marine habitats. Of the 99 non-marine species, 49 were from terrestrial habitats and 50 were aquatic, especially the aquatic stages of Diptera (44 species). The abundance of records from species of Aedes,Ochlerotatus and Psorophora contrasts markedly with a paucity of records from species of Anopheles,Culex and Culiseta. Records from terrestrial isopods are numerous (19 species), although the diversity of IIVs that infect them is mostly unstudied. IIV infections have been reported from every continent, except Antarctica, but there are few records from Africa, southern Asia and Latin America. Most reports describe patent IIV infections as rare whereas inapparent (covert) infection may be common in certain species. The relationship between particle size and iridescent colour of the host is found to be consistent with optical theory in the great majority of cases. Only 24 reported IIVs from insect hosts have partial characterization data and only two have been subjected to complete genome sequencing. I show that the rate of publication on IIVs has slowed from 1990 to the present, and I draw a number of conclusions and suggestions from the host list and make recommendations for future research efforts.

Los virus iridiscentes de invertebrados (VIIs) son virus icosaedrales de ADN que infectan a invertebrados, principalmente insectos e isópodos terrestres en hábitats húmedos y acuáticos. Búsquedas extensivas de bases de datos resultaron en la identificación de 79 artículos científicos, los cuales reportaron 108 especies de invertebrados infectados naturalmente por iridovirus. De estos, 103 (95%) fueron artrópodos y los otros fueron moluscos, un anélido y un nematodo. Nueve especies fueron de hábitats marinos. De las 99 especies no marinas, 49 fueron terrestres y 50 fueron acuáticas, especialmente los estadios acuáticos de dípteros (44 especies). La abundancia de infecciones en especies de Aedes,Ochlerotatus y Psorophora se contrasta marcadamente con la escasez de casos en especies de Anopheles,Culex y Culiseta. Reportes de infecciones de los isópodos terrestres son numerosos (19 especies), aunque la diversidad de los VII que los infectan es desconocida. Se han reportado infecciones por VIIs de todos los continentes, excepto Antártica, pero se notan pocos ejemplos de África, Asia y Latinoamérica. La mayoría de los artículos señala que las infecciones patentes son poco comunes, mientras que las infecciones enmascaradas (subletales) pueden ser comunes en algunas especies. La relación entre el tamaño de la partícula y el color iridiscente concuerda con la teoría óptica en casi todos los casos. Veinticuatro de los VIIs de insectos han sido caracterizados parcialmente y solo dos de éstos han sido secuenciados completamente. Demuestro que el ritmo de publicación sobre los VIIs ha disminuido en los últimos 15 años, señalo varias conclusiones y sugerencias de la lista de especies de huéspedes y presento algunas recomendaciones para la investigación futura con este grupo de patógenos.