Immunization of TAA-derived peptides induced peptide-specific and HLA-DR4-restricted Th-cell responses in Tgm.

<p>Peptides emulsified in CFA and IFA were injected into the tail base of Tgm or C57BL/6 mice (WT) on day 0 and day 7. On day 14, the splenocytes were harvested and cultured <i>in </i><i>vitro</i> for 7 days with the peptides (1 μg/ml). (A) Purified CD4⁺ cells (1 × 10⁵/well) were co-cultured with L-DR4 (3 × 10⁴/well) pulsed with or without CMV-egH_290–302 peptide for 72 h and ³H-thymidine uptake was measured. (B) Purified CD4⁺ cells (1 × 10⁵/well) were co-cultured with L cells (3 × 10⁴/well) or L-DR4 (3 × 10⁴/well) pulsed with or without CDCA1_55-78 peptide for 72 h and ³H-thymidine uptake was measured. (C, D) Immunization of Tgm with syngeneic BM-DCs (5 × 10⁵) pulsed with WT1_332-347 peptide (C) or KIF20A_494-517 peptide (D) followed by a booster shot of the peptide in CFA successfully induced IFN-γ production by Th cells (1 × 10⁵/well) in response to the peptide-pulsed L-DR4 cells (5 × 10⁴/well), but not to unpulsed L-DR4 cells.