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. Author manuscript; available in PMC: 2014 Oct 1.
Published in final edited form as: Virus Res. 2013 Jul 23;177(1):10.1016/j.virusres.2013.07.008. doi: 10.1016/j.virusres.2013.07.008

Figure 5. Effect of murine ß-defensin 2 (Mbd2) on the frequency of HA-518 and NP-147 epitope-specific IFN-γ secreting CD8+ T cells in the spleens of HAd-HA-NP vaccinated aged mice.

Figure 5

Aged mice (18 month-old) were inoculated intramuscularly (i.m.) either with PBS or 1 × 108 plaque-forming units (pfu) of HAd-Mbd2 (adenovirus vector expressing Mbd2). Seven days later, animals were immunized with 1 × 108 pfu of HAd-HA-NP [adenovirus vector expressing hemagglutinin (HA) and nucleoprotein (NP) of a H5N1 influenza virus]. Animals similarly inoculated with 1 × 108 pfu of HAd-ΔE1E3 on days 0 and 7 to serve as negative controls. Mice were euthanized four weeks after the last immunization, and the spleens were collected. Single cell suspensions were prepared by passage through screens, and 1 × 106 cells were cultured in the presence of HA-518 or NP-147 peptide on anti-interferon-γ antibody-coated 96-well filter plates and developed according to an ELISpot protocol. The ELISpot plates were read using Bioreader 5000 (BIOSYS Miami, FL). The bars represent mean ± standard deviation (SD) from 3 animals/group. ** ; P<0.001, *** ; P<0.0001, compared to HAd-ΔE1E3 or PBS-HAd-HA-NP control group.