Enhanced Immunogenicity of Peptide P277 by Heat Shock Protein HSP65 Vector Carrying Tandem Repeats of P277 to Prevent Type 1 Diabetes in NOD Mice

Abstract

The peptide P277 contains a target epitope for diabetogenic T cells and it has been used as an ideal target antigen to develop vaccines against type 1 diabetes. A major problem in developing P277 vaccine is its low immunogenicity. Recent applications involving multiple copies of self-peptide in linear alignment and conjugation with carrier proteins appear to increase the immune response. In this study, we designed a method based on isocaudamer technique to repeat tandemly the 24-residue sequence P277, then 6 tandemly repeated copies of the peptide P277 were fused to mycobacterial heat-shock protein 65 to construct a fusion protein HSP65-6×P277 as an immunogen. We examined the effect of the tandem repeats of the peptide P277 in eliciting an immune response by comparing the immunogenicity of the three immunogens: P277, HSP65-P277 and HSP65-6×P277. Immunization of mice with the fusion protein HSP65-6×P277 elicited much higher levels of specific anti-P277 antibodies than with P277 and HSP65-P277, which should suggest that multiple tandem repeats of a certain epitope is an efficient method to overcome the low immunogenicity of self-peptide antigens and the immunogen HSP65-6×P277 might be further developed to a vaccine against type 1 diabetes.

Key words

HSP65 - P277 - type 1 diabetes - tandem repeat epitope - immunogenicity

References

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