Characterisation of the antibody response to a totally synthetic immunocontraceptive peptide vaccine based on LHRH.

In this study we describe our attempts to improve the immunogenicity of a synthetic epitope-based vaccine. The vaccine consists of an epitope (P25) that is recognised by CD4+ helper T cells and the target epitope luteinising hormone releasing hormone (LHRH). We show that replacement of the single cysteine residue within P25 with amino acids such as alanine, aminobutyric acid, serine or with carboxymethylated cysteine leads to diminished immunogenicity of the vaccine and only the oxidised dimeric form of the peptide retains the full immunogenicity of the vaccine. Secondly, by measuring the serum antibody response and the number of the antigen secreting cells in spleen and bone marrow we found that three doses of 20 nmol per mouse induced the more consistent and higher immune responses than those induced by three doses of either 2 nmol or 80 nmol per mouse. A greater variation in antibody titre was observed in mice that received the 2 mol or 80 nmol dose regimes. Last, by administering the vaccine in its lipidated form in the presence or absence of additional adjuvant we found that either inoculation regime elicited similar antibody responses. Only at low doses of antigen was a synergistic effect observed when lipopeptide was co-administered with additional adjuvant.