The goal of this proposal is to localize by a synthetic approach the 'continuous' antigenic sites of the Beta subunit of human major histocompatibility antigen HLA-DR2. The work is a collaboration between Z. Atassi (Baylor) and P. Cresswell (Duke University). The DR2 Beta subunits will be purified from a DR2 homozygous B-Lymphoblastoid cell line and will be used to raise rabbit and mouse antisera and to prepare DR2 Beta-specific rat and mouse monoclonal antibodies. These antisera, together with a panel of human alloantisera that are either specific to, or cross-reactive with, HLA-DR2 will be used to localize the antigenic sites on the DR2 Beta subunit. We shall employ the synthetic strategy we recently introduced for the localization of continuous antigenic sites in proteins. The approach consists of the synthesis of a series of consecutive overlapping peptides that together represent the entire primary structure of the protein. Thirty overlapping (5-residue overlaps) peptides representing the entire extracellular part of DR2 Beta subunit, the intracellular piece and the various sequences reported in the region 60-69 will be synthesized. The peptides will be studied for their ability to bind the aforementioned polyclonal and monoclonal antibodies. The peptides will also be used to determine the interacting surfaces on the DR2 Beta-subunit with the Alpha- and I-subunits. The approach will narrow the antigenic sites to within 9-13 residues. Further synthesis will be carried out around these indicated regions to achieve narrower delineation of the antigenic sites. Polyclonal and monoclonal antibodies will be prepared against immunochemically active, as well as other, peptides and will be studied for their ability to bind HLA-DR2 and DR2 Beta subunit. These studies will enable better understanding, in molecular terms of the role HLA-DR antigens in immune responses.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI021386-04
Application #
3131449
Study Section
Experimental Immunology Study Section (EI)
Project Start
1984-07-01
Project End
1990-05-31
Budget Start
1987-07-01
Budget End
1989-05-31
Support Year
4
Fiscal Year
1987
Total Cost
Indirect Cost
Name
Baylor College of Medicine
Department
Type
Schools of Medicine
DUNS #
074615394
City
Houston
State
TX
Country
United States
Zip Code
77030
Lewis, D E; Ulrich, R G; Atassi, H et al. (1990) HLA-DR peptide inhibits HIV-induced syncytia. Immunol Lett 24:127-31
Ulrich, R G; Atassi, M Z (1990) Mapping of the full profile of T cell allorecognition regions on HLA-DR2 beta subunit. Eur J Immunol 20:713-21
Ulrich, R G; Atassi, M Z (1989) Alloreactive T cell recognition of the HLA-DR beta N-terminal polymorphic region. Immunol Lett 21:285-90
Atassi, M Z; Manshouri, T; Yokoi, T (1988) Recognition of inter-transmembrane regions of acetylcholine receptor alpha subunit by antibodies, T cells and neurotoxins. Implications for membrane-subunit organization. FEBS Lett 228:295-300
Atassi, M Z; McDaniel, C S; Manshouri, T (1988) Mapping by synthetic peptides of the binding sites for acetylcholine receptor on alpha-bungarotoxin. J Protein Chem 7:655-66
Ulrich, R G; Atassi, H; Lutz, P et al. (1987) Immune recognition of human major histocompatibility antigens: localization by a comprehensive synthetic strategy of the continuous antigenic sites in the first domain of HLA-DR2 beta chain. Eur J Immunol 17:497-502
Atassi, H; Ulrich, R G; Atassi, M Z (1987) The continuous antigenic regions in the second domain of the beta chain of human MHC DR2 antigen: antigenic profile of the entire extracellular part of the chain. Eur J Immunol 17:769-73
Mulac-Jericevic, B; Atassi, M Z (1986) Segment alpha 182-198 of Torpedo californica acetylcholine receptor contains second toxin-binding region and binds anti-receptor antibodies. FEBS Lett 199:68-74
McCormick, D J; Lennon, V A; Atassi, M Z (1985) Synthesis of an antigenic site of native acetylcholine receptor peptide 159-169 of Torpedo acetylcholine receptor alpha-chain. Biochem J 226:193-7
Lennon, V A; McCormick, D J; Lambert, E H et al. (1985) Region of peptide 125-147 of acetylcholine receptor alpha subunit is exposed at neuromuscular junction and induces experimental autoimmune myasthenia gravis, T-cell immunity, and modulating autoantibodies. Proc Natl Acad Sci U S A 82:8805-9