Figure 3

Testing of individual ORFs comprising parapoxvirus library pool X identifies the B2L gene. The ORF-expressing LEEs from pool X were inoculated individually into mouse abdomen sites using the gene gun (1 μg LEE per site), in triplicate, and skin sections were obtained 4 days later. Samples were processed and immuno-stained for the MHC II surface marker I-A^d/I-E^d with antibody 2G9. Scale bars are 200 micron. A. Immunostains of tissue sections from ORFV ORF-expressing LEEs comprising pool X. Viral gene numbers are indicated alongside corresponding skin sections. Each of the eight representative tissue sections is presented as an overlay of FITC labeled anti-I-A^d/I-E^d fluorescent microscopy. B. The same section of skin inoculated with B2L was viewed under both white visible (wh) light to observe gold particles (left panel) and under fluorescence (fluor) to visualize FITC-labeled class II positive cells (I-A/I-E+) (middle panel). An untreated section of skin was stained with FITC labeled anti-I-A/I-E and visualized under fluorescence (fluor) (right panel).