Functional Characterization of Antibodies against Neisseria gonorrhoeae Opacity Protein Loops

Background

The development of a gonorrhea vaccine is challenged by the lack of correlates of protection. The antigenically variable neisserial opacity (Opa) proteins are expressed during infection and have a semivariable (SV) and highly conserved (4L) loop that could be targeted in a vaccine. Here we compared antibodies to linear (Ab_linear) and cyclic (Ab_cyclic) peptides that correspond to the SV and 4L loops and selected hypervariable (HV₂) loops for surface-binding and protective activity in vitro and in vivo.

Methods/Findings

Ab_{SV cyclic} bound a greater number of different Opa variants than Ab_{SV linear}, including variants that differed by seven amino acids. Antibodies to the 4L peptide did not bind Opa-expressing bacteria. Ab_{SV cyclic} and Ab_{HV2 cyclic}, but not Ab_{SV linear} or Ab_{HV2 linear} agglutinated homologous Opa variants, and Ab_{HV2BD cyclic} but not Ab_{HV2BD linear} blocked the association of OpaB variants with human endocervical cells. Only Ab_{HV2BD linear} were bactericidal against the serum resistant parent strain. Consistent with host restrictions in the complement cascade, the bactericidal activity of Ab_{HV2BD linear} was increased 8-fold when rabbit complement was used. None of the antibodies was protective when administered vaginally to mice. Antibody duration in the vagina was short-lived, however, with <50% of the antibodies recovered 3 hrs post-administration.

Conclusions

We conclude that an SV loop-specific cyclic peptide can be used to induce antibodies that recognize a broad spectrum of antigenically distinct Opa variants and have agglutination abilities. HV₂ loop-specific cyclic peptides elicited antibodies with agglutination and adherence blocking abilities. The use of human complement when testing the bactericidal activity of vaccine-induced antibodies against serum resistant gonococci is also important.