title: In silico prediction of Ebola Zaire GP(1,2) immuno-dominant epitopes for the Balb/c mouse.
abstract: BACKGROUND
  Ebola is a Filovirus (FV) that induces a highly communicable and deadly hemorrhagic fever. Currently, there are no approved vaccines to treat FV infections. Protection from FV infection requires cell mediated and humoral immunity. Glycoprotein GP(1,2) Fc Zaire, a recombinant FV human Fc fusion protein, has been shown to confer protection against mouse adapted Zaire Ebola virus. The present studies are focused upon identifying immunodominant epitopes using in silico methods and developing tetramers as a diagnostic reagent to detect cell mediated immune responses to GP(1,2) Fc.
  
  
  METHODS
  The GP(1,2) Ebola Zaire sequence from the 1976 outbreak was analyzed by both BIMAS and SYFPEITHI algorithms to identify potential immuno-dominant epitopes. Several peptides were synthesized and screened in flow-based MHC stability studies. Three candidate peptides, P8, P9 and P10, were identified and, following immunization in Balb/c mice, all three peptides induced IFN-γ as detected by ELISpot and intracellular staining.
  
  
  RESULTS
  Significantly, P8, P9 and P10 generated robust cytotoxic T-cell responses (CTL) as determined by a flow cytometry-based Caspase assay. Antigen specific cells were also detected, using tetramers. Both P9 and P10 have sequence homology with highly conserved regions of several strains of FV.
  
  
  CONCLUSIONS
  In sum, three immunodominant sequences of the Ebola GP(1,2) have been identified using in silico methods that may confer protection against mouse adapted Ebola Zaire. The development of tetramer reagents will provide unique insight into the potency and durability of medical countermeasure vaccines for known bioterrorism threat agents in preclinical models.
authors.affiliation: Division of Biology, Chemistry and Materials Science, Center for Devices and Radiological Health, FDA, Silver Spring, MD, 20993, USA. debargh.dutta@gmail.com.|Department of Medicine, USUHS, 4301 Jones Bridge road, Bethesda, MD, 20814, USA. debargh.dutta@gmail.com.
authors.forename: Debargh K
authors.initials: DK
authors.lastname: Dutta
authors.affiliation: Division of Biology, Chemistry and Materials Science, Center for Devices and Radiological Health, FDA, Silver Spring, MD, 20993, USA. krhodes2@terpmail.umd.edu.|University of Maryland, College Park, MD, 20742, USA. krhodes2@terpmail.umd.edu.
authors.forename: Kelly
authors.initials: K
authors.lastname: Rhodes
authors.affiliation: Division of Biology, Chemistry and Materials Science, Center for Devices and Radiological Health, FDA, Silver Spring, MD, 20993, USA. Steven.Wood@fda.hhs.gov.
authors.forename: Steven C
authors.initials: SC
authors.lastname: Wood
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D003198:Computer Simulation; D029043:Ebolavirus / Q000276:immunology*; D058501:Enzyme-Linked Immunospot Assay; D005260:Female; D006649:Histocompatibility Antigens / Q000276:immunology; D007114:Immunization; D016056:Immunodominant Epitopes / Q000737:chemistry / Q000276:immunology*; D007371:Interferon-gamma / Q000378:metabolism; D042541:Intracellular Space / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry / Q000276:immunology; D055503:Protein Multimerization; D016415:Sequence Alignment; D013154:Spleen / Q000166:cytology; D013194:Staining and Labeling; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology
chemical_list: D006649:Histocompatibility Antigens; D016056:Immunodominant Epitopes; D010455:Peptides; D007371:Interferon-gamma