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Differential Gene Expression Pattern of Importin β3 and NS5 in C6/36 Cells Acutely and Persistently Infected with
María Leticia Ávila-Ramírez1, Ana Laura Reyes-Reyes2, Rodolfo Gamaliel Avila-Bonilla3
1Doctorado en Ciencias en Biotecnología, Escuela Nacional de Medicina y Homeopatía, Instituto Politécnico Nacional, Mexico City 07320, Mexico.
Abstract:
The establishment of persistent dengue virus infection within the cells of the mosquito vector is an essential requirement for viral transmission to a new human host. The mechanisms involved in the establishment and maintenance of persistent infection are not well understood, but it has been suggested that both viral and cellular factors might play an important role. In the present work, we evaluated differential gene expression in Aedes albopictus cells acutely (C6/36-HT) and persistently infected (C6-L) with Dengue virus 2 by cDNA-AFLP. We observed that importin β3 was upregulated in noninfected cells compared with C6-L cells. Using RT-qPCR and plaque assays, we observed that Dengue virus levels in C6-L cells essentially do not vary over time, and peak viral titers in acutely infected cells are observed at 72 and 120 h postinfection. The expression level of importin β3 was higher in acutely infected cells than in persistently infected cells; this correlates with higher levels of NS5 in the nucleus of the cell. The differential pattern of importin β3 expression between acute and persistent infection with Dengue virus 2 could be a mechanism to maintain viral infection over time, reducing the antiviral response of the cell and the viral replicative rate.
Insights
Persistent dengue virus infection in mosquito cells relies on the regulation of importin β3. This study reveals importin β3 is lower in persistent infections, potentially reducing viral replication and host antiviral responses.
Area of Science:
- Virology
- Molecular Biology
- Entomology
Background:
- Persistent dengue virus infection in mosquito vectors is crucial for disease transmission.
- Mechanisms maintaining persistent dengue virus infections are not fully understood.
- Both viral and cellular factors are implicated in persistent infections.
Purpose of the Study:
- To investigate differential gene expression in Aedes albopictus cells during acute and persistent Dengue virus 2 infection.
- To identify cellular factors involved in maintaining persistent dengue virus infections.
Main Methods:
- cDNA-AFLP (Complementary DNA-Amplified Fragment Length Polymorphism) was used to analyze gene expression.
- RT-qPCR (Reverse Transcription Quantitative Polymerase Chain Reaction) quantified viral RNA levels.
- Plaque assays determined viral titers in infected cells.
Main Results:
- Importin β3 expression was upregulated in uninfected cells compared to persistently infected cells (C6-L).
- Dengue virus levels remained stable in persistently infected cells.
- Higher importin β3 expression in acutely infected cells correlated with increased nuclear NS5 protein levels.
Conclusions:
- Differential expression of importin β3 may be a key mechanism for maintaining persistent dengue virus infections.
- Reduced importin β3 in persistent infections could lower cellular antiviral responses and viral replication rates.
- Understanding these mechanisms is vital for controlling dengue transmission by mosquito vectors.
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