The anchor-modification is needed to induce effector CTLs capable of degranulation. ABabDII mice were immunized subcutaneously with (A) the native peptide TEL-AML1 or (B) the anchor-modified peptide TEL-AML1-9V, mixed with CpG oligodeoxynucleotides, PBS and IFA. They were boosted, and splenocytes and draining LNs were isolated 10 days later. The cells were in vitro restimulated for 6 hours with 10−6M specific peptides (TEL-AML1 or TEL-AML1-9V), with an irrelevant peptide (NY-BR-1), or with anti-CD3/CD28 antibodies, as indicated. Prestaining for CD107a was performed by including anti-CD107a antibodies during the whole period of stimulation, with the addition of monensin. The cells were subsequently stained for CD3 and CD8 and analyzed by flow cytometry. Events shown are gated on CD3+CD8+ lymphocytes; numbers indicate percentages of cells in the respective quadrants. One representative of 5 (A) and 1 of 6 (B) mice are shown, from 2 independent experiments with similar results.
