title: Structural mutants of dengue virus 2 transmembrane domains exhibit host-range phenotype.
abstract: BACKGROUND
  There are over 700 known arboviruses and at least 80 immunologically distinct types that cause disease in humans. Arboviruses are transmitted among vertebrates by biting insects, chiefly mosquitoes and ticks. These viruses are widely distributed throughout the world, depending on the presence of appropriate hosts (birds, horses, domestic animals, humans) and vectors. Mosquito-borne arboviruses present some of the most important examples of emerging and resurgent diseases of global significance.
  
  
  METHODS
  A strategy has been developed by which host-range mutants of Dengue virus can be constructed by generating deletions in the transmembrane domain (TMD) of the E glycoprotein. The host-range mutants produced and selected favored growth in the insect hosts. Mouse trials were conducted to determine if these mutants could initiate an immune response in an in vivo system.
  
  
  RESULTS
  The DV2 E protein TMD defined as amino acids 452SWTMKILIGVIITWIG467 was found to contain specific residues which were required for the production of this host-range phenotype. Deletion mutants were found to be stable in vitro for 4 sequential passages in both host cell lines. The host-range mutants elicited neutralizing antibody above that seen for wild-type virus in mice and warrant further testing in primates as potential vaccine candidates.
  
  
  CONCLUSIONS
  Novel host-range mutants of DV2 were created that have preferential growth in insect cells and impaired infectivity in mammalian cells. This method for creating live, attenuated viral mutants that generate safe and effective immunity may be applied to many other insect-borne viral diseases for which no current effective therapies exist.
authors.affiliation: Arbovax, Incorporated, Raleigh, North Carolina, USA. ksmith@arbovax.com
authors.forename: Katherine M
authors.initials: KM
authors.lastname: Smith
authors.forename: Kavita
authors.initials: K
authors.lastname: Nanda
authors.forename: Carla J
authors.initials: CJ
authors.lastname: Spears
authors.forename: Mariana
authors.initials: M
authors.lastname: Ribeiro
authors.forename: Ricardo
authors.initials: R
authors.lastname: Vancini
authors.forename: Amanda
authors.initials: A
authors.lastname: Piper
authors.forename: Gwynneth S
authors.initials: GS
authors.lastname: Thomas
authors.forename: Malcolm E
authors.initials: ME
authors.lastname: Thomas
authors.forename: Dennis T
authors.initials: DT
authors.lastname: Brown
authors.forename: Raquel
authors.initials: R
authors.lastname: Hernandez
mesh_terms: D000330:Aedes; D000595:Amino Acid Sequence; D000818:Animals; D057134:Antibodies, Neutralizing / Q000097:blood; D000914:Antibodies, Viral / Q000097:blood; D002460:Cell Line; D003716:Dengue Virus / Q000235:genetics / Q000276:immunology / Q000472:pathogenicity / Q000502:physiology*; D058507:Host Specificity; D051379:Mice; D008807:Mice, Inbred BALB C; D008958:Models, Molecular; D008969:Molecular Sequence Data; D050505:Mutant Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D017384:Sequence Deletion; D014759:Viral Envelope Proteins / Q000235:genetics* / Q000276:immunology / Q000378:metabolism*
chemical_list: D057134:Antibodies, Neutralizing; D000914:Antibodies, Viral; C051821:E-glycoprotein, Dengue virus type 2; D050505:Mutant Proteins; D014759:Viral Envelope Proteins