Figure 5.
CTL epitopes can be processed and presented to elicit high frequency responses. (A) HLA-A2 transgenic mice were immunized on days 0, 7 and 14 with an ImmunoBody™ construct containing either HLA-A*0201 restricted gp100 210M (n = 12), gp100 wild type (n = 6), Mage3 (n = 6), hTERT (n = 6) or Tie2 (n = 6) epitopes in CDRH1. On day 19 splenocytes were analyzed by IFNγ elispot assay against gp100 210M, gp100 wild type, Mage3, hTERT or Tie2 peptides and an irrelevant control. Responses are measured as spots/million splenocytes. (B) C57Bl/6, Balb/c or HLA-A*0201 transgenic mice were immunized on days 0, 7 and 14 with an ImmunoBody™ construct containing the H-2Kb restricted TRP2 epitope (n = 50) which is also restricted through HLA-A*0201 (n = 12), the H-2Kd restricted HepB epitope (n = 6), the H-2Kb Ovalbumin epitope (n = 6) or the HLA-A*0201 restricted hTERT epitope (n = 6) in CDRH2. On day 19 splenocytes were assayed ex vivo in IFNγ elispot assay against relevant and control peptides. Responses are measured as spots/million splenocytes. (C) C57Bl/6 or HLA-A*0201 transgenic mice were immunized on days 0, 7 and 14 with an ImmunoBody™ construct containing the H-2Kb restricted TRP2 epitope (n = 12) or HLA-A*0201 restricted VEGFR2 epitopes (n = 6) in CDRH3. On day 19 splenocytes were assayed for presence of specific responses by IFNγ elispot assay. Responses are measured as spots/million splenocytes. (D) Splenocytes from immunized mice were depleted of CD8 T cells and analyzed against TRP2 peptide, HepB helper peptide and a media control in triplicate for the presence epitope specific responses in IFNγ elispot assay in triplicate. Responses are measured as spots/million splenocytes (n = 4). (E) HLA-A*0201 transgenic mice were immunized on days 0, 7 and 14 with an ImmunoBody™ construct containing HLA-A*0201 restricted TRP2 epitope in CDRH2, the HLA-A*0201 restricted gp100 210M epitope in CDRH1 and the I-Ab restricted HepB helper epitope in CDRL1. On day 19 splenocytes were analyzed by IFNγ elispot assay against relevant peptides and an irrelevant control in triplicate. Responses are measured as spots/million splenocytes (n = 12). All results are an average of at least two independent experiments.