Figure 3.

Cloning of the TCRα and TCRβ chain from mB29b-TCR hybridoma LHEP4 into TCR⁻ cells results in Hsp70 peptide-specific transfectants. Cloned transfected cells were cocultured with A20 B cells (A) or BMDCs (B), which were loaded with 5, 0.5, or 0.05 μg/ml Hsp70 peptides. After stimulation with Hsp70 peptides mB29a, mB29b, or B29, as well as length variants of mB29b (a1a or a8), supernatants were added to CTLL-16 cells and IL-2-dependent proliferation was determined after 24 h. The graph shows more IL-2-dependent proliferation of CTLL-16 cells when stimulated with a high concentration Hsp70 peptide mB29b or B29. There is no response to stimulation with mB29a and to a minor extent to the length variants of mB29b (a8 or a1a). Similar results were obtained with the BMDC coculture (B). As a control, cocultured LHEP4 cells were unstimulated or stimulated with ConA (data not shown). ³H-thymidine incorporation is shown as mean of triplicate samples/well ± SEM. Data shown are representative of two independent experiments.