Characterization and Transferring of Human Rotavirus Double-Layered Particles in MA104 Cells.
BACKGROUND
Rotavirus (RV) is a major cause of gastroenteritis in infants and children and is one of the most severe public health problems. Rotaviruses outer layer contains two proteins including VP4 and VP7. These proteins are necessary for host-cell binding and penetration. TLP (triple layer virus particle) of RV is a complete infectious virion that binds to the target cells and internalized at the cytoplasm. The DLP (double layer virus particle) is a non-infectious particle that is formed through exclusion of the outer layer proteins including VP4 and VP7. These DLPs are the transcriptionally active forms of rotavirus.


OBJECTIVE
The aim of this study was to transfer DLP of RV into cytoplasm of MA104 cells by Lipofectamine and to analyze their replication.


METHODS
Initially, rotavirus was purified by CsCl discontinuous gradient and DLP was separated from TLP based on density differences. For confirmation, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) of the proteins were conducted Then the purified DLP of RV was transferred into MA104 cells using Lipofectamine.


RESULTS
We attempt to avoid the attachment and entry of the rotavirus by using Lipofectamine to mediate the delivery of viral particles directly into the cytoplasm. DLP was endocytosed into the cytoplasm following treatment by Lipofectamine and then replicated in cytoplasm.


CONCLUSIONS
Therefore the non-infectious DLPs were became infectious if introduced into the cytoplasm of permissive and cancerous cells, without passing attachment and entry process.
Department of Virology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, IR Iran.
Ali
A
Teimoori
Department of Virology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, IR Iran.
Hoorieh
H
Soleimanjahi
Department of Virology, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, IR Iran.
Manoochehr
M
Makvandi
Rotavirus; Transfection; Viral Plaque Assay