HCMV epitope-specific T cell response following primary and secondary immunisation with Ad-CMVpoly vaccine.

<p>Two different groups of HHD-2 mice were immunised intramuscularly with Ad-CMVpoly (7.5×10⁸ PFU/mouse). <i>A & B</i>, Following primary immunisation, animals were sacrificed 10 days post immunisation and HCMV epitope-specific reactivity was assessed in the splenocytes by ELISPOT assays as described in the “<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0003256#s4" target="_blank">Material and Methods</a>” section. The epitopes tested for T cell reactivity were VLE (IE-1), NLV (pp65), RIF (pp65), VLA (IE-1), IIY (IE-2) AVG (gB). <i>C & D</i>, For immunological analysis following secondary immunisation, animals were given booster immunisation (7.5×10⁸ PFU/mouse) 100 days after primary immunisation and then sacrificed 10 days post secondary immunisation. HCMV epitope-specific reactivity was assessed as described above. <i>A & C</i> shows ELISPOT data based on the pooled HLA A2-restricted HCMV epitopes, while <i>B & D</i> shows relative T cell responses to individual epitopes. The results are expressed as Mean±SE of spot forming cells (SFC) per 10⁶ splenocytes from four individually tested mice. <i>E,</i> Anti-adenovirus antibody titre induced by immunisation with Ad-CMVpoly. Serum samples were collected at different time points after immunisation and anti-adenovirus titres were evaluated by ELISA as described in the “<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0003256#s4" target="_blank">Material and Methods</a>” section. All statistical analyses were conducted using GraphPad Prism 4 software.