Identification of hepatitis A virus mimotopes by phage display, antigenicity and immunogenicity.
A phage-displayed peptide approach was used to identify ligands mimicking antigenic determinants of hepatitis A virus (HAV) for the first time. Bacteriophages displaying HAV mimotopes were isolated from a phage-display peptide library by affinity selection on serum antibodies from hepatitis A patients. Selected phage-peptides were screened for reactivity with sera from HAV infected patients and healthy controls. Four cloned peptides with different sequences were identified as mimotopes of HAV; three of them showed similarity in their amino acid sequences with at least one of the VP3 and VP1 antigenic proteins of HAV. One clone was recognised by 92% of the positive sera. The phagotopes competed effectively with HAV for absorption of anti-HAV-specific antibodies in human sera, as determined by ELISA. The four phage clones induced neutralising anti-HAV antibodies in immunised mice. These results demonstrate the potential of this method to elucidate the disease related epitopes of HAV and to use these mimotopes in diagnostic applications or in the development of a mimotope-based hepatitis A vaccine without the necessity of manipulation of the virus.
Larralde
Osmany G
OG
Institute of Biomedical and Life Sciences, University of Glasgow, Glasgow, UK. 0202767L@student.gla.ac.uk
Martinez
Raiza
R
Camacho
Frank
F
Amin
Nevis
N
Aguilar
Alicia
A
Talavera
Arturo
A
Stott
David I
DI
Perez
Ela M
EM
D000595:Amino Acid Sequence; D000818:Animals; D001435:Bacteriophages / Q000235:genetics / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D035921:Hepatitis A Antigens / Q000737:chemistry / Q000276:immunology*; D017957:Hepatitis A Virus, Human / Q000235:genetics / Q000276:immunology*; D006801:Humans; D007106:Immune Sera / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D018716:Molecular Mimicry / Q000276:immunology*; D008969:Molecular Sequence Data; D009500:Neutralization Tests; D019151:Peptide Library; D010455:Peptides / Q000276:immunology / Q000302:isolation & purification; D017386:Sequence Homology, Amino Acid
D035921:Hepatitis A Antigens; D007106:Immune Sera; D019151:Peptide Library; D010455:Peptides