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

Figure 4.

Figure 4

Linked help is required for optimal CTL responses. (A) C57Bl/6 or HLA-A*0201 transgenic mice were immunized at day 0, 7 and 14 with ImmunoBody™ heavy chain only DNA constructs containing the H-2Kb restricted TRP2 epitope in CDRH2 or the HLA-A*0201 restricted gp100 210M epitope in CDRH1. On day 19, splenocytes were analyzed by IFNγ elispot assay against gp100 210M peptide, TRP2 peptide and control. Responses are measured as spots/million splenocytes (n = 6). (B) C57Bl/6 mice were immunized at days 0, 7 and 14 with an ImmunoBody™ DNA containing the H-2Kb restricted TRP2 epitope in CDRH2, the HLA-A*0201 restricted epitope gp100 210M in CDRH1 and the I-Ab restricted HepB helper epitope in CDRL1 with either human IgG1 or murine IgG2a constant domains. On day 19, splenocytes were analyzed by IFNγ elispot assay against TRP2 peptide, HepB helper peptide and control. Responses are measured as spots/million splenocytes (n = 6). (C) HLA-DR*0401 transgenic mice were immunized at days 0, 7 and 14 with an ImmunoBody™ DNA containing the gp100 DR4 epitope in CDR H1, TRP2 epitope in CDR H2 and gp100 DR7 epitope in CDR H3 with either human IgG1 or murine IgG2a constant domains. On day 19, splenocytes were analyzed by IFNγ elispot assay against TRP2 peptide, gp100 DR4 helper peptide and control. Responses are measured as spots/million splenocytes (n = 6). (D) C57Bl/6 (n = 48), Balb/c (n = 9) or HLA-DR*0401 (n = 6) transgenic mice were immunized on days 0, 7 and 14 with an ImmunoBody™ construct containing either I-Ab restricted HepB, I-Ad restricted Influenza or HLA-DR*0401 restricted gp100 epitopes in CDR L1. On day 19 splenocytes were analyzed by IFNγ elispot assay against HepB, Influenza or gp100 helper peptides and an irrelevant control. Responses are measured as spots/million splenocytes. (E) C57Bl/6 (n = 6), HLA-DR*0101 (n = 6) or HLA-DR*0401 (n = 6) transgenic mice were immunized on days 0, 7 and 14 with an ImmunoBody™ construct containing either I-Ab restricted HepB or HLA-DR*0401 restricted gp100 or tyrosinase or HLADR* 0101 restricted triosephosphate isomerase (TPI) epitopes in CDR L3. On day 19 splenocytes were analyzed by IFNγ elispot assay against HepB, gp100, TPI or tyrosinase helper peptides and an irrelevant control. Responses are measured as spots/million splenocytes. All results are an average of at least two independent experiments.