The assembly and immunological properties of non-linear synthetic immunogens containing T-cell and B-cell determinants.
For the rational design of synthetic vaccines, a potential immunogen must contain the appropriate helper T-cell and B-cell determinants to elicit a strong and relevant immune response. In this study we describe a method for the assembly of antigenic determinants from influenza virus hemagglutinin onto a lysine-based support, resulting in dimeric and trimeric constructs bearing both T-cell and B-cell determinants. A panel of synthetic immunogens was constructed incorporating peptides representing: (i) the B-cell determinant TLKLATG and the T-cell determinant PKYVKQNTLKLA which overlaps this sequence in the heavy chain (HA1) of the hemagglutinin; and (ii) the same B-cell determinant with an alternate T-cell determinant ALNNRFQIKGVELKS from the light chain (HA2). With these peptides we were able to investigate the effects of altering the source of T-cell help, increasing the copy number of B-cell determinants as well as comparing the presentation of determinants in either linear tandem or branched geometries. In general, peptides incorporating the non-native helper T-cell determinant in a branched conformation were superior immunogens, eliciting higher titres of both peptide-specific and virus-specific antibody. Increasing the copy number of the B-cell determinant also proved to be an advantage in terms of increasing antibody titres. Other evidence was obtained indicating that presentation of determinants to T cells may be different for linear peptide constructs compared to branched immunogens bearing the same determinants.
Fitzmaurice
C J
CJ
Department of Microbiology, University of Melbourne, Parkville, Victoria, Australia.
Brown
L E
LE
McInerney
T L
TL
Jackson
D C
DC
D000595:Amino Acid Sequence; D000818:Animals; D000914:Antibodies, Viral / Q000096:biosynthesis; D004306:Dose-Response Relationship, Immunologic; D018985:Epitopes, B-Lymphocyte / Q000737:chemistry* / Q000276:immunology*; D018984:Epitopes, T-Lymphocyte / Q000737:chemistry* / Q000276:immunology*; D005260:Female; D006389:Hemagglutinins, Viral / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D008808:Mice, Inbred CBA; D008969:Molecular Sequence Data; D009975:Orthomyxoviridae / Q000276:immunology; D010455:Peptides / Q000276:immunology
D000914:Antibodies, Viral; D018985:Epitopes, B-Lymphocyte; D018984:Epitopes, T-Lymphocyte; D006389:Hemagglutinins, Viral; D010455:Peptides