Transcriptome Analysis of an Aedes albopictus Cell Line Single- and Dual-Infected with Lammi Virus and WNV

Understanding the flavivirus infection process in mosquito hosts is important and fundamental in the search for novel control strategies that target the mosquitoes’ ability to carry and transmit pathogenic arboviruses. A group of viruses known as insect-specific viruses (ISVs) has been shown to interfere with the infection and replication of a secondary arbovirus infection in mosquitoes and mosquito-derived cell lines. However, the molecular mechanisms behind this interference are unknown. Therefore, in the present study, we infected the Aedes albopictus cell line U4.4 with either the West Nile virus (WNV), the insect-specific Lammi virus (LamV) or an infection scheme whereby cells were pre-infected with LamV 24 h prior to WNV challenge. The qPCR analysis showed that the dual-infected U4.4 cells had a reduced number of WNV RNA copies compared to WNV-only infected cells. The transcriptome profiles of the different infection groups showed a variety of genes with altered expression. WNV-infected cells had an up-regulation of a broad range of immune-related genes, while in LamV-infected cells, many genes related to stress, such as different heat-shock proteins, were up-regulated. The transcriptome profile of the dual-infected cells was a mix of up- and down-regulated genes triggered by both viruses. Furthermore, we observed an up-regulation of signal peptidase complex (SPC) proteins in all infection groups. These SPC proteins have shown importance for flavivirus assembly and secretion and could be potential targets for gene modification in strategies for the interruption of flavivirus transmission by mosquitoes.

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Expression profile of the PIWI mRNAs protein family in human cells experimentally infected with Dengue Virus 4

Research Society and Development ◽

10.33448/rsd-v10i3.13371 ◽

2021 ◽

Vol 10 (3) ◽

pp. e32010313371

Author(s):

Walter Félix Franco Neto ◽

Murilo Tavares Amorim ◽

Karla Fabiane Lopes de Melo ◽

Gustavo Moraes Holanda ◽

Jardel Fábio Lopes Ferreira ◽

...

Keyword(s):

Viral Load ◽

Viral Replication ◽

Infection Process ◽

Messenger Rnas ◽

Altered Expression ◽

Hemorrhagic Fevers ◽

Laboratory Results ◽

Albopictus Cell ◽

Infected Cells ◽

Hepatocyte Cells

Objective: Evaluate the messenger RNAs (mRNA) PIWI proteins expression during infection with VDEN 4 in human hepatocyte cells. Materials and Methods: VDEN4 strain H778494 (JQ513335) was used, which was stored in Aedes albopictus cell culture (Clone C6 / 36) at the Arbovirology and Hemorrhagic Fevers section of the Evandro Chagas Institute. The techniques of cell cultures, stock (C6/36), inoculation, extraction, viral load quantification, RTqPCR and statistical analyzes were performed at the Viral Biogenesis Laboratory. Results: According to the results obtained, two cells (HepG2 and Huh7.5) demonstrated a higher level of viral replication at 72 hours post infection (hpi). The PIWI 2 target alter its mRNA expression during VDEN4 infection. The PIWI 4 target expression, was observed an altered expression in the infected cells. Thus, it was found that they are in different poles, while the viral load in the first three days showed high expression. The expression of mRNA was low in relation to the normal rate given by the uninfected cells. Conclusion: In our findings, it was observed that PIWI2 and 4 proteins have an inverse relationship to the viral infection process by VDEN4, when there is an increase in viral replication, these two proteins end up having a significant reduction in their expression. Probably, this reduction of expression is involved with the biogenesis process of apoptosis-regulating microRNAs.

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Small RNA Response to Infection of the Insect-Specific Lammi Virus and Hanko Virus in an Aedes albopictus Cell Line

Viruses ◽

10.3390/v13112181 ◽

2021 ◽

Vol 13 (11) ◽

pp. 2181

Author(s):

Pontus Öhlund ◽

Juliette Hayer ◽

Jenny C. Hesson ◽

Anne-Lie Blomström

Keyword(s):

Cell Line ◽

Aedes Albopictus ◽

Small Rna ◽

Phylogenetic Groups ◽

Ping Pong ◽

Sequence Elements ◽

Albopictus Cell ◽

Infected Cells ◽

Interfering Rna ◽

Unknown Sequence

RNA interference (RNAi)-mediated antiviral immunity is believed to be the primary defense against viral infection in mosquitoes. The production of virus-specific small RNA has been demonstrated in mosquitoes and mosquito-derived cell lines for viruses in all of the major arbovirus families. However, many if not all mosquitoes are infected with a group of viruses known as insect-specific viruses (ISVs), and little is known about the mosquito immune response to this group of viruses. Therefore, in this study, we sequenced small RNA from an Aedes albopictus-derived cell line infected with either Lammi virus (LamV) or Hanko virus (HakV). These viruses belong to two distinct phylogenetic groups of insect-specific flaviviruses (ISFVs). The results revealed that both viruses elicited a strong virus-derived small interfering RNA (vsiRNA) response that increased over time and that targeted the whole viral genome, with a few predominant hotspots observed. Furthermore, only the LamV-infected cells produced virus-derived Piwi-like RNAs (vpiRNAs); however, they were mainly derived from the antisense genome and did not show the typical ping-pong signatures. HakV, which is more distantly related to the dual-host flaviviruses than LamV, may lack certain unknown sequence elements or structures required for vpiRNA production. Our findings increase the understanding of mosquito innate immunity and ISFVs’ effects on their host.

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Analysis of the miRNA expression profile in an Aedes albopictus cell line in response to bluetongue virus infection

Infection Genetics and Evolution ◽

10.1016/j.meegid.2016.01.012 ◽

2016 ◽

Vol 39 ◽

pp. 74-84 ◽

Cited By ~ 15

Author(s):

Shanshan Xing ◽

Junzheng Du ◽

Shandian Gao ◽

Zhancheng Tian ◽

Yadong Zheng ◽

...

Keyword(s):

Virus Infection ◽

Cell Line ◽

Aedes Albopictus ◽

Expression Profile ◽

Mirna Expression ◽

Bluetongue Virus ◽

Mirna Expression Profile ◽

Albopictus Cell

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A06 Inhibition of West Nile virus infection on Aedes albopictus cell line, C6/36 : The role of JNK signaling pathway on virus infection

Medical Entomology and Zoology ◽

10.7601/mez.54.24_2 ◽

2003 ◽

Vol 54 (supplement) ◽

pp. 24

Author(s):

T. Mizutani ◽

M. Kobayashi ◽

Y. Eshita ◽

K. Shirato ◽

Y. Ako ◽

...

Keyword(s):

West Nile Virus ◽

Virus Infection ◽

Cell Line ◽

Signaling Pathway ◽

Aedes Albopictus ◽

West Nile Virus Infection ◽

Jnk Signaling ◽

Albopictus Cell ◽

Jnk Signaling Pathway

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Transfection and colonization of Tetranychus truncatus Wolbachia strain wTtru in cell lines of the mosquito Aedes albopictus

Systematic and Applied Acarology ◽

10.11158/saa.23.12.11 ◽

2018 ◽

Vol 23 (12) ◽

pp. 2420 ◽

Cited By ~ 2

Author(s):

Xue Xia ◽

Chang-Wu Peng ◽

Yi-Jia Lu ◽

Xiao-Ying Zheng ◽

Xiao-Yue Hong

Keyword(s):

Phylogenetic Analysis ◽

Success Rate ◽

Cell Line ◽

Aedes Albopictus ◽

Wolbachia Strain ◽

Dynamic Changes ◽

Endosymbiotic Bacteria ◽

Production Practice ◽

Albopictus Cell ◽

The Relationship

Wolbachia are endosymbiotic bacteria that commonly infect arthropods and cause reproductive manipulations in hosts. Artificial transfection is one of the best methods for the study of Wolbachia-induced reproductive manipulations and its application in production practice. However, the low success rate of artificial transfection laid burden for investigation. The success rate of artificial transfection of Wolbachia between closely related hosts is usually higher than distant hosts, but the mechanism of transfection is unknown. In our study, the wTtru strain (supergroup B) infecting the Arachnida Tetranychus truncatus was transfected into the Aedes albopictus cell line (native-infected with wAlB strain from supergroup B). The wTtru strain was quantified and localized in the cell line to observe dynamic changes after transfection. A phylogenetic analysis of the relationship between the wTtru strain and the wAlB strain showed that the wTtru (ST=278) strain belonged to supergroup B and was closely related to the mosquitoes native Wolbachia wAlbB (ST=464), so we speculate that successful transfection was related to the Wolbachia strain, and furthermore, Wolbachia strain may be more important than host relationship. Our results provide a factual basis for studying artificial transfection of Wolbachia, and show that Wolbachia transfection between distant hosts can be achieved.

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