The objective is to obtain detailed information concerning the structure of hepatitis B viral proteins, and to determine the relationship between their protein structure and their antigenic function. These studies will include research on the HBsAg, including the pre-S gene products, HBeAg, HBcAg and the """"""""X"""""""" protein. Studies of the HBsAg will include studies of the intact particle and its protein and lipid components. We will utilize our panel of monoclonal anti-HBs antibodies (which contain anti-group specific and anti-subtypes specific antibodies) in conjunction with amino acid sequence analysis, chemical modification studies, synthetic peptides, immunoblotting techniques, olignucleotide directed site, specific mutagenesis, and eukaryotic expression vectors for the production of HBsAg from the cloned viral gene to attempt to determine the physical structure of HBsAg and the location and structure of the various antigenic determinants. We will also use physical techniques such as freeze ETCH and freeze fracture electron microscopy low angle x-ray scattering, fourier transform infrared spectorphotometry, and circular dichroism to gain information on the number, arrangement, and secondary structure of the protein subunits with HBsAg and how the lipid components are related to the maintenance of this structure. The role of the pre-S proteins in the particle structure and antigenic activity will also be investigated. The chemistry of the conversion of HBcAg to HBeAg will be studied and this chemical conversion correlated with the antigenic alteration. The location of the relevant antigenic sites will be sought by the same methods as used for HBsAg. We will also use anti-synthetic peptide antibodies in an attempt to characterize the product of the viral """"""""X"""""""" gene. Mammalian expression vectors will be used in an effort to synthesize the protein for further characterization. The antibody produced in humans in response to HBV infection or to immunization by the current vaccine (Heptavax) as well as that produced in response to the experimental yeast HBsAg vaccines will be examined by competition studies with our monoclonal antibodies to determine the relative amounts and specificities of these human antibodies. It is expected that this will help define the major immunogenic determination recognized by humans.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
2R01AI015955-07A1
Application #
3126505
Study Section
Allergy and Immunology Study Section (ALY)
Project Start
1979-08-01
Project End
1989-07-31
Budget Start
1986-08-01
Budget End
1987-07-31
Support Year
7
Fiscal Year
1986
Total Cost
Indirect Cost
Name
Virginia Commonwealth University
Department
Type
Overall Medical
DUNS #
City
Richmond
State
VA
Country
United States
Zip Code
23298
Yelamos, B; Nunez, E; Gomez-Gutierrez, J et al. (2001) Urea equilibrium unfolding of the major core protein of the retrovirus feline immunodeficiency virus and its tryptophan mutants. Biochim Biophys Acta 1546:87-97
Nath, M D; Peterson, D L (2001) In vitro assembly of feline immunodeficiency virus capsid protein: biological role of conserved cysteines. Arch Biochem Biophys 392:287-94
Nunez, E; Wei, X; Delgado, C et al. (2001) Cloning, expression, and purification of histidine-tagged preS domains of hepatitis B virus. Protein Expr Purif 21:183-91
Rodriguez-Crespo, I; Yelamos, B; Albar, J P et al. (2000) Selective destabilization of acidic phospholipid bilayers performed by the hepatitis B virus fusion peptide. Biochim Biophys Acta 1463:419-28
Rodriguez-Crespo, I; Nunez, E; Yelamos, B et al. (1999) Fusogenic activity of hepadnavirus peptides corresponding to sequences downstream of the putative cleavage site. Virology 261:133-42
Yelamos, B; Nunez, E; Gomez-Gutierrez, J et al. (1999) Circular dichroism and fluorescence spectroscopic properties of the major core protein of feline immunodeficiency virus and its tryptophan mutants. Assignment of the individual contribution of the aromatic sidechains. Eur J Biochem 266:1081-9
Birkett, A J; Yelamos, B; Rodriguez-Crespo, I et al. (1997) Cloning, expression, purification, and characterization of the major core protein (p26) from equine infectious anemia virus. Biochim Biophys Acta 1339:62-72
Rodriguez-Crespo, I; Gomez-Gutierrez, J; Encinar, J A et al. (1996) Structural properties of the putative fusion peptide of hepatitis B virus upon interaction with phospholipids. Circular dichroism and Fourier-transform infrared spectroscopy studies. Eur J Biochem 242:243-8
Jin, L; Wei, X; Gomez, J et al. (1995) Use of alpha-N,N-bis[carboxymethyl]lysine-modified peroxidase in immunoassays. Anal Biochem 229:54-60
Milich, D R; Peterson, D L; Schodel, F et al. (1995) Preferential recognition of hepatitis B nucleocapsid antigens by Th1 or Th2 cells is epitope and major histocompatibility complex dependent. J Virol 69:2776-85

Showing the most recent 10 out of 23 publications