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De Novo Design of Peptide Immunogens That Mimic the Coiled Coil Region of Human T-cell Leukemia Virus Type-1 Glycoprotein 21 Transmembrane Subunit for Induction of Native Protein Reactive Neutralizing Antibodies
Ist Teil von
The Journal of biological chemistry, 2004-06, Vol.279 (23), p.24141-24151
Ort / Verlag
United States: American Society for Biochemistry and Molecular Biology
Erscheinungsjahr
2004
Quelle
EZB-FREE-00999 freely available EZB journals
Beschreibungen/Notizen
Peptide vaccines able to induce high affinity and protective neutralizing antibodies must rely in part on the design of antigenic
epitopes that mimic the three-dimensional structure of the corresponding region in the native protein. We describe the design,
structural characterization, immunogenicity, and neutralizing potential of antibodies elicited by conformational peptides
derived from the human T-cell leukemia virus type 1 (HTLV-1) gp21 envelope glycoprotein spanning residues 347â374. We used
a novel template design and a unique synthetic approach to construct two peptides (WCCR2T and CCR2T) that would each assemble
into a triple helical coiled coil conformation mimicking the gp21 crystal structure. The peptide B-cell epitopes were grafted
onto the ϵ side chains of three lysyl residues on a template backbone construct consisting of the sequence acetyl-XGKGKGKGCONH2
(where X represents the tetanus toxoid promiscuous T cell epitope (TT) sequence 580â599). Leucine substitutions were introduced
at the a and d positions of the CCR2T sequence to maximize helical character and stability as shown by circular dichroism and guanidinium
hydrochloride studies. Serum from an HTLV-1-infected patient was able to recognize the selected epitopes by enzyme-linked
immunosorbent assay (ELISA). Mice immunized with the wild-type sequence (WCCR2T) and the mutant sequence (CCR2T) elicited
high antibody titers that were capable of recognizing the native protein as shown by flow cytometry and whole virus ELISA.
Sera and purified antibodies from immunized mice were able to reduce the formation of syncytia induced by the envelope glycoprotein
of HTLV-1, suggesting that antibodies directed against the coiled coil region of gp21 are capable of disrupting cell-cell
fusion. Our results indicate that these peptides represent potential candidates for use in a peptide vaccine against HTLV-1.