Directing the immune response to carbohydrate antigens.
Peptide mimetics may substitute for carbohydrate antigens in vaccine design applications. At present, the structural and immunological aspects of antigenic mimicry, which translate into immunologic mimicry, as well as the functional correlates of each, are unknown. In contrast to screening peptide display libraries, we demonstrate the feasibility of a structure-assisted vaccine design approach to identify functional mimeotopes. By using concanavalin A (ConA), as a recognition template, peptide mimetics reactive with ConA were identified. Designed peptides were observed to compete with synthetic carbohydrate probes for ConA binding, as demonstrated by enzyme-linked immunosorbent assay and isothermal titration calorimetry (ITC) analysis. ITC measurements indicate that a multivalent form of one particular mimetic binds to ConA with similar affinity as does trimannoside. Splenocytes from mimeotope-immunized mice display a peptide-specific cellular response, confirming a T-cell-dependent nature for the mimetic. As ConA binds to the Envelope protein of the human immunodeficiency virus, type 1 (HIV-1), we observed that mimeotope-induced serum also binds to HIV-1-infected cells, as assessed by flow cytometry, and could neutralize T-cell line adapted HIV-1 isolates in vitro, albeit at low titers. These studies emphasize that mimicry is based more upon functional rather than structural determinants that regulate mimeotope-induced T-dependent antibody responses to polysaccharide and emphasize that rational approaches can be employed to develop further vaccine candidates.
Cunto-Amesty
G
G
Department of Pathology and Laboratory Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104, USA.
Dam
T K
TK
Luo
P
P
Monzavi-Karbassi
B
B
Brewer
C F
CF
Van Cott
T C
TC
Kieber-Emmons
T
T
D000595:Amino Acid Sequence; D000818:Animals; D000941:Antigens / Q000737:chemistry*; D001665:Binding Sites; D001667:Binding, Competitive; D001671:Biochemistry / Q000379:methods*; D002151:Calorimetry; D002240:Carbohydrate Sequence; D002241:Carbohydrates / Q000737:chemistry*; D002455:Cell Division; D002469:Cell Separation; D003208:Concanavalin A / Q000737:chemistry* / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004797:Enzyme-Linked Immunosorbent Assay; D018604:Epitope Mapping; D005434:Flow Cytometry; D007700:Kinetics; D008024:Ligands; D051379:Mice; D008807:Mice, Inbred BALB C; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D017386:Sequence Homology, Amino Acid; D013154:Spleen / Q000166:cytology; D013816:Thermodynamics; D014612:Vaccines / Q000737:chemistry
D000941:Antigens; D002241:Carbohydrates; D008024:Ligands; D010455:Peptides; D014612:Vaccines; D003208:Concanavalin A