The Generation of Human Hepatoma Cells (Huh7) Bearing Self Replicating Norwalk Virus RNA

Reference Number: 06-19

Inventor: Kyeong-Ok Chang

Background

Norwalk virus continues to be a leading cause of gastrointestinal problems affecting the cruise ship industry and militaries around the world. Norwalk virus is a prototype of the noroviruses in the Caliciviridae family that is associated with acute gastroenteritis outbreaks. Recent studies showed that noroviruses are responsible for greater than 90% of non-bacterial gastroenteritis outbreaks and are associated with an estimated 23 million cases of gastroenteritis in the U.S. alone each year. These viruses are highly infectious with the potential to be deliberately disseminated in food or water supplies as an agent of bio-terrorism.

Research at Kansas State University has produced a novel cell replicon that can be used to grow the virus in cell culture for vaccine production. The novelty of this invention pertains to the ability to produce the cell replicon in a human cell line.

This invention describes a method for generation of cells expressing self-replicating norovirus RNA molecules (norovirus “replicons”) in a human cell line (Huh-7). It will facilitate the development of vaccines and therapeutic drugs for norovirus and perhaps other calicivirus diseases.

Because noroviruses could not be grown in cell culture, research on vaccine and antiviral development has been severly hampered (if not impossible). Previously, there were no stable cell culture based systems to screen potential antiviral drugs against noroviruses and caliciviruses.

Application

Practical applications, variations, and potential extent of possible uses include but are not limited to:

  • Method which generates any calicivirus replicon harboring cells using any cell lines
  • Method which screens antiviral drug development using calicivirus replicon harboring cells
Advantages

The generation of additional replicon harboring cells with a human hepatoma cell line (Huh7 cells) was a significant achievement because of host specificity and wide availability of commercial reagents for the human cell line. In addition, Huh7 cell based replicon produced much higher levels of viral proteins than BHK21 based replicon. Compared to BHK cells, the replion in Huh7 cells produce higher efficiency in the replication and is more stable. Using the replicon harboring cells, interferon alpha effectively inhibited the replication of Norwalk virus replicon in the cells, and proved the possibility of the use of this replicon harboring cell line to screen antiviral drug development.

Patent Status
  • Patent protection is not being pursued because it is our belief that this should be used as a research tool for vaccine production and drug development. The novel cell replicon is offered on a non-exclusive basis to all interested parties for a one-time upfront fee.

Kansas State University Research Foundation seeks to have discussions with companies that are interested in licensing and/or research collaborations.

Interested parties should contact:

Kansas State University Institute for Commercialization (KSU-IC)
2005 Research Park Circle Manhattan, KS 66502
Tel: 785-532-3900 Fax: 785-532-3909
E-Mail: ic@k-state.edu