Immunization onto bare skin with synthetic peptides: immunomodulation with a CpG‐containing oligodeoxynucleotide and effective priming of influenza virus‐specific CD4+ T cells

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Abstract
Exploiting the immune system of the skin for vaccine administration offers an attractive alternative to the currently used invasive immunization procedures. In this study we report that a synthetic peptide representing a T-helper (Th) epitope from influenza virus haemagglutinin (aa 307–319) can be an effective immunogen when coapplied with cholera toxin (CT) onto bare skin. Proliferation of both peptide- and influenza virus-specific CD4+ T cells was measured in lymphocyte cultures from spleens and regional lymph nodes. The presence of the CpG oligodeoxynucleotide 1826 in the peptide/CT formulation, enhanced the proliferation of peptide- and virus-specific T cells as measured by the conventional [3H]thymidine uptake and interleukin (IL)-2 assays. Furthermore, the bias towards Th2-type of responses stimulated by CT was shifted towards Th1 as demonstrated (i) by the increase of interferon-γ and decrease of IL-4 cytokine levels measured in culture supernatants, (ii) by the predominance of IG2a anti-CT antibodies in the serum, and (iii) by the down-regulation of total serum IgE antibody levels. These findings demonstrate the potential of the bare skin as a non-invasive route for administration of small molecular size peptide antigens. Furthermore, with the selection and combination of the appropriate type of adjuvants, immune responses can be modulated towards the desired type of Th phenotype.