Analysis of the Function of Nonstructural Genes and T Cell epitopes of Mouse Hepatitis Virus Targeted Recombination

Mouse hepatitis virus - JHM strain (MHV-JHM) causes an acute encephalitis and a virus-induced demyelinating disease. This mouse disease has similarities to the human demyelinating disease multiple sclerosis. The ability to clone genes into a coronavirus has been recently developed by Masters et al who has developed a recombinant MHV-A59 virus. Our lab is interested in creating a recombinant MHV-JHM. We obtained a chimeric virus that is approximately 75% MHV-JHM and 25% MHV-A59. We initially replaced the 25% of the chimeric virus that is A59 with the homologous sequences from MHV-JHM. With the creation of a recombinant MHV-JHM, we will be able to study the in vivo effects of specific mutations in structural and nonstructural genes. Our lab has previously identified the important CD8 and CD4 epitopes which are found within the S gene. Virus specifically mutated in these epitopes will supply clinical profiles of epitope function. Gene 4 is known to encode nonstructural protein ns4 whose function remains unknown. Previous studies have been able to look at the growth of gene 4 mutated virus in cell cultures but this project will extend those studies to in vivo analysis of the function of gene 4. We will be able to test these recombinant virus in tissue culture and our mouse model of acute encephalitis.

Ontiveros E, Kim TS, Gallagher TM, Perlman S. Enhanced virulence mediated by the murine coronavirus, mouse hepatitis virus strain JHM, is associated with a glycine at residue 310 of the spike glycoprotein. J Virol. 2003 Oct;77(19):10260-9. PubMed PMID: 12970410; PubMed Central PMCID: PMC228498.