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. 2009 Jan-Feb;1(1):71–85. doi: 10.4161/mabs.1.1.7492

Figure 3.

Figure 3

ImmunoBody™ DNA immunization. (A) C57Bl/6 mice were immunized at days 0, 7 and 14 with an ImmunoBody™ protein containing the H-2Kb epitope in the CDRH3 site (n = 6) or the equivalent DNA (intact) (n = 12) or ImmunoBody™ DNA containing the H-2Kb restricted TRP2 epitope in CDRH2 (non associated; n = 9). On day 19 splenocytes from immunized mice were analyzed ex vivo for the presence of epitope specific responses. Responses are measured as spots/million splenocytes. (B) C57Bl/6 mice were immunized on days 0, 7 and 14 with an ImmunoBody™ DNA construct containing TRP2 epitope in CDRH2 with or without leader sequence. On day 19 splenocytes were analyzed by IFNγ elispot assay against TRP2 peptide in triplicate. Responses are measured as spots/million splenocytes and normalised against an irrelevant peptide control (n = 4). (C) C57Bl/6 mice were immunized at days 0, 7 and 14 with an ImmunoBody™ DNA containing the H-2Kb epitope in the CDRH2 site (n = 6) or whole murine TRP2 antigen in pcDNA3 (n = 6) or pOrig vectors (n = 6). On day 19 splenocytes from immunized mice were analyzed ex vivo for the presence of epitope specific responses. Responses are measured as spots/million splenocytes. (D) C57Bl/6 mice were immunized with human IgG1 DNA containing TRP2 epitope in CDRH2 at days 0 and 7 (n = 6), or 0, 7 and 14 (n = 6). Splenocytes were analyzed on day 19 for the presence of TRP2 epitope specific responses by IFNγ elispot. Responses are measured as spots/million splenocytes. (E) C57Bl/6 mice were immunized with ImmunoBody™ DNA at days 0, 7 and 14 via gene gun (n = 6). Responses were analyzed at day 20, day 48 post primary immunization. A parallel group of mice was boosted at day 42 (n = 6). Responses were tested in ex vivo elispot assay against TRP2 peptide and an irrelevant control in triplicate. Responses are measured as spots/million splenocytes. All results are an average of two independent experiments.