Induction of double restricted responses after peptide immunization of HLA transgenic mice.

<p>Eighteen groups of 3–5 mice were injected intraperitoneally three times at two week intervals with CAF09 plus one of the 9-11mer peptides (20nM). One week after the last immunization, IFNγ ELISpot was used to measure HLA-A*02:01 and/or HLA-DRB1*01:01 restricted CD8 and CD4 T cell responses. As a control, a group of mice (n = 4) was injected intraperitoneally three times at two week intervals with CAF09 plus the known HLA-A*02:01 binding peptide M(58–66) (21). Each bar represents mean ± SD of IFNγ spot forming units (SFU) of groups of 3–5 immunized mice from which splenocytes were pooled and depleted for CD4 or CD8 positive cells before being stimulated <i>ex vivo</i> with peptide in four replicate cultures. Experiments were conducted 1–5 times depending on the individual peptides.