Murine B-cell and T-cell epitopes of the allergen Hev b 5 from natural rubber latex.
Hev b 5, a proline-rich acidic protein with a predominantly random secondary structure, is a major allergen in natural rubber latex and a candidate for specific immunotherapy of latex allergy. As a first step in the identification of candidate peptides for immunotherapy, we have begun to identify the B-cell and T-cell epitopes of Hev b 5 in BALB/c mice. The mice were immunized with a Hev b 5 fusion protein. The B-cell epitopes were determined by the SPOTS method using overlapping octamers or by ELISA inhibition using a series of overlapping 20-mers. The T-cell epitopes were determined by the proliferation and cytokine release of splenocytes cultured in the presence of the 20-mers. Potential antibody binding regions included residues in regions 1-38, 55-74, 109-128 and 132-151. Examination of the binding sequences for common motifs suggested enhanced antibody binding to the KXEE or KEXE sequences, where X is empty, threonine or alanine. Splenocyte stimulation and cytokine release suggest T-cell epitopes with the regions 1-20, 37-56, 73-101 and 109-146. Since they may contain major T-cell epitopes but do not exhibit significant antibody binding, peptide regions 38-48 and 75-101 are candidates for specific immunotherapy to Hev b 5 in the BALB/c mouse model.
Slater
J E
JE
Center for Molecular Mechanisms of Disease Research, Children's Research Institute, Children's National Medical Center, Washington, DC 20010, USA. slaterj@cber.fda.gov
Paupore
E J
EJ
O'Hehir
R E
RE
D000485:Allergens / Q000235:genetics / Q000276:immunology*; D000595:Amino Acid Sequence; D000818:Animals; D017951:Antigen Presentation; D052179:Antigens, Plant; D001402:B-Lymphocytes / Q000276:immunology*; D000939:Epitopes / Q000235:genetics / Q000276:immunology*; D020315:Latex Hypersensitivity / Q000276:immunology*; D008211:Lymphocyte Cooperation; D051379:Mice; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000235:genetics / Q000276:immunology; D010940:Plant Proteins; D017421:Sequence Analysis; D013601:T-Lymphocytes / Q000276:immunology*
D000485:Allergens; D052179:Antigens, Plant; D000939:Epitopes; D010446:Peptide Fragments; D010940:Plant Proteins; C101800:allergen Hev b 5