Identification of critical residues of an immunodominant region of Echinococcus granulosus antigen B.
Immune evasion strategies often shape the immunogenicity of parasite components. We recently found that the N-terminal extension of the major subunit of Echinococcus granulosus antigen B (AgB), the causative agent of hydatid disease, concentrates the immunoreactive B cell epitopes of the native molecule. The nature of this immunodominance was analyzed using four monoclonal antibodies (mAbs) defining overlapping epitopes in this region of the AgB molecule. The minimal epitope requirements of these mAbs were determined using phage display peptide libraries. The consensus sequences isolated with the mAbs, and alanine replacement analysis with synthetic peptides mapped the relevant molecular contacts within a short stretch corresponding to residues 17-24 of the AgB major subunit. Substitution of two critical residues within this stretch produced a dramatic loss of antigenicity, as determined by using patient sera. The circular dichroism spectra of the antigen, together with the distribution of the contact residues, suggest that this region adopts an amphipathic alpha-helix structure that clusters the contact residues on its polar side. To provide further insight in the interpretation of the structure activity relationships for this immunoreactive region of E. granulosus AgB, we developed a model for the N-terminal extension of the AgB major subunit, which helps to rationalize our data.
González-Sapienza
Gualberto
G
Cátedra de Inmunología, Facultad de Química, Universidad de la República, Instituto de Higiene, Av. A. Navarro 3051, Montevideo 11600, Uruguay. ggonzal@fg.edu.uy
Cachau
Raul E
RE
D000595:Amino Acid Sequence; D000818:Animals; D004446:Echinococcus / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D015801:Helminth Proteins; D008074:Lipoproteins / Q000737:chemistry*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D017386:Sequence Homology, Amino Acid
D015801:Helminth Proteins; D008074:Lipoproteins; C089273:antigen B, Echinococcus