Supporting Figures

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Supporting Figure 5
Supporting Figure 6
Supporting Figure 7
Supporting Figure 8

Fig. 5. Increasing the number of DCs used to immunize mice increases the T cell response but competition still occurs. CFSE-labeled P14.Thy1.1 were injected with or without OT-I cells, and mice were immunized with various numbers of DCs as shown. Spleen cells were gated on CD8⁺ class II^- cells. P14 cells are identified by Thy1.1 expression, and OT-I cells are identified by staining with K^b-Ova-PE tetramers. This is the same experiment in as Fig. 1.

Fig. 6. OT-I cells divide to a lesser extent in the presence of P14 cells. A total of 1 ´ 10⁷ OT-I.Thy1.1 spleen cells labeled with CFSE were injected i.v. in the presence or absence of equal numbers of P14 spleen cells. These mice were immunized 24 h later with 2 ´ 10⁶ DCs pulsed with no antigen, ovalbumin and gp33 peptides, or A2 (SAINFEKL, a weak agonist version of the ovalbumin peptide) and gp33. OT-I responses were measured on day 4 after immunization. Histograms are gated on CD8⁺ Thy1.1⁺ class II MHC^- events.

Fig. 7. Additional activation marker expression on P14 cells at 48 h in the presence or absence of OT-I competition. Mice were injected i.v. with CFSE-labeled P14 cells and OT-I cells and peptide-pulsed DCs as shown, and P14 cells in the spleen were tested for activation marker expression at 48 h as in Fig. 2.

Fig. 8. Transfer of competing OT-I cells at the time of immunization results in competition. Cell transfer and immunization were performed as in previous experiments except that OT-I cells were transferred at the time points indicated. DC immunization took place at 0 h. A representative of two experiments is shown.