Prime/boost vaccination route impacts vaginal mucosal antibody levels and antigen reactive B cells.

<p>Mucosal antigen-specific IgG (<b>a</b>) and IgA (<b>b</b>) titres from vaginal mucosal lavage were assessed (n = 8 per group) at the end of the study (week 13). Antibody results are expressed as group means (µg/ml+SEM). At the end of the study, antibody secreting IgG (<b>c</b>) and IgA (<b>d</b>) cells were obtained from excised spleens and plated at a density of 5×10⁶ cells/ml in a commercial IgG or IgA ELISPOT plate. Samples were stimulated with 5 µg/ml gp140 antigen for 16 h and developed as per the manufacturer’s instructions. Results represent group means (n = 8 mice per group) of IgG or IgA Spot Forming Units (SFU)/million antigen stimulated cells (+ SEM). Antigen-specific IgA secreting B cells from enzymatically digested and mechanically disrupted vaginal tissue (<b>e</b>) were determined using a direct ELISPOT assay at the end of the study (week 13). Results represent pooled samples and are expressed as fold increase in SFU/million antigen stimulated cells (+ SEM) as compared to negative controls. Statistical significance was assessed using Mann Whitney U test with ***<i>p</i><0.0005, **<i>p</i><0.005 and *<i>p</i><0.05. Data are representative of two independent experiments or are pooled from two experiments.