Verification of gene expression by the H1N1 vaccine candidates.

<p>HA expression in A549 cells transduced with AdHA.wt, AdHA.cod or Ad5. <b>A,</b> HA detection in cell lysates by western blot analysis (60 ug of total protein loaded in each lane) using ferret antiserum. <b>B</b>, Flow cytometric analysis of HA expression at the cell surface using ferret or mouse antiserum. Shown are the percentages of HA positive cells.</p

Induced protection against H1N1 by the H1N1 vaccine candidates.

<p>Protection of immunized animals (5 or 6 weeks after immunization with AdHA.wt vs Ad5 control or AdHA.cod vs Ad5 control) against an intranasal challenge with 1000 pfu of A/Ohio/7/09 (H1N1)pdm was measured as viral titers in lung and nasal turbinate determined 3 days post-inoculation in a plaque formation assay using MDCK-L cells. Shown are log10 values of mean titer for each group ± SEM. The horizontal dashed line represents the lower limit of detection of the assay.</p

Induction of humoral and cellular immune responses by the H1N1 vaccine candidates.

<p>Induction of influenza H1N1-specific immune responses after subcutaneous immunization of Balb/c mice with adenovirus encoding wildtype (AdHA.wt) or codon-optimized (AdHA.cod) HA antigen or control adenovirus (Ad5). <b>A,</b> Antibody to influenza A/Texas/05/09 (H1N1)pdm in sera of mice after receiving dose escalations of the vaccine candidates (AdHA.wt or AdHA.cod). Shown are log2 values of HI-titer. Horizontal lines represent geometric means in each group. <b>B</b>, Induction of H1N1-specific neutralizing antibodies in sera 5 weeks after immunization with AdHA.wt, AdHA.cod or Ad5 control. Shown are neutralizing antibody titer to influenza A/Texas/05/09 (H1N1)pdm measured by microneutralization assay. Horizontal lines represent geometric means in each group. <b>C,</b> HA peptide specific T cell responses in splenocytes detected in IFNγ Elispot assays. A representative result is shown as means of SFC ± SEM of triplicate determinations in each group.</p

Protection against influenza A/Ohio/7/09 (H1N1)pdm.

(1)<p>number of animals from which virus was cultured/number of animals inoculated.</p