A structure-based approach to a synthetic vaccine for HIV-1.
The generation of neutralizing antibodies by peptide immunization is dependent on achieving conformational compatibility between antibodies and native protein. Consequently, approaches are needed for developing conformational mimics of protein neutralization sites. We replace putative main-chain hydrogen bonds (NH --> O=CRNH) with a hydrazone link (N-N=CH-CH(2)CH(2)) and scan constrained peptides for fit with neutralizing monoclonal antibodies (MAbs). To explore this approach, a V3 MAb 58.2 that potently neutralizes T-cell lab-adapted HIV-1(MN) was used to identify a cyclic peptide, [JHIGPGR(Aib)F(D-Ala)GZ]G-NH(2) (loop 5), that binds with >1000-fold higher affinity than the unconstrained peptide. NMR structural studies suggested that loop 5 stabilized beta-turns at GPGR and R(Aib)F(D-Ala) in aqueous solvent implying considerable conformational mimicry of a Fab 58.2 bound V3 peptide determined by X-ray crystallography [Stanfield, R. L. et al. (1999) Structure 142, 131-142]. Rabbit polyclonal antibodies (PAbs) generated to loop 5 but not to the corresponding uncyclized peptide bound the HIV-1(MN) envelope glycoprotein, gp120. When individual rabbit antisera were scanned with linear and cyclic peptides, further animal-to-animal differences in antibody populations were characterized. Loop 5 PAbs that most closely mimicked MAb 58.2 neutralized HIV-1(MN) with similar potency. These results demonstrate the remarkable effect that conformation can have on peptide affinity and immunogenicity and identify an approach that can be used to achieve these results. The implications for synthetic vaccine and HIV-1 vaccine research are discussed.
Cabezas
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The Burnham Institute, 10901 Torrey Pines Road, La Jolla, California 92037, USA.
Wang
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M
Parren
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PW
Stanfield
R L
RL
Satterthwait
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AC
D016915:AIDS Vaccines / Q000138:chemical synthesis / Q000737:chemistry* / Q000276:immunology; D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism; D001666:Binding Sites, Antibody; D001667:Binding, Competitive / Q000276:immunology; D002999:Clone Cells; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D015699:HIV Envelope Protein gp120 / Q000008:administration & dosage / Q000276:immunology / Q000378:metabolism; D015497:HIV-1 / Q000276:immunology*; D006801:Humans; D006825:Hybridomas; D051379:Mice; D018716:Molecular Mimicry; D008969:Molecular Sequence Data; D019906:Nuclear Magnetic Resonance, Biomolecular; D009842:Oligopeptides / Q000008:administration & dosage / Q000138:chemical synthesis / Q000276:immunology / Q000378:metabolism; D010456:Peptides, Cyclic / Q000008:administration & dosage / Q000138:chemical synthesis / Q000276:immunology / Q000378:metabolism; D011817:Rabbits; D013329:Structure-Activity Relationship; D014614:Vaccines, Synthetic / Q000737:chemistry* / Q000276:immunology
D016915:AIDS Vaccines; D000911:Antibodies, Monoclonal; D015699:HIV Envelope Protein gp120; D009842:Oligopeptides; D010456:Peptides, Cyclic; D014614:Vaccines, Synthetic