Figure 7.

LAG3 expression on CD8⁺ T cells increases with length of antigenic stimulation. (A) Transgenic OT1 splenocytes were stimulated for either 24 h or 72 h with a fixed concentration of SIINFEKL peptide and assayed for LAG3 expression on 4-1BB⁺ CD8⁺ T cells by flow cytometry at 72 h (data is representative of three independent experiments, with 5–10 independent samples each). (B) Transgenic OT1 splenocytes were adoptively transferred into wild-type C57BL6 mice (n = 4 to 5 animals per group) and administered a priming immunization of SIINFEKL peptide (100 µg) and pTVG4 (50 µg) 24 h later. Four days after priming, animals were divided into groups and boosted with either PBS, SIINFEKL peptide (100 µg), or SIINFEKL peptide (100 µg) and pTVG4 (50µg). Two days after the booster immunization, levels of LAG3 were assayed on SIINFEKL tetramer⁺ CD8⁺ T cells from the spleens of treated mice by flow cytometry. (C) Transgenic OT1 splenocytes were stimulated with either SIINFEKL, EIINFEKL, or SIIGFEKL peptides at 2 µg/mL for 24 h and assayed for LAG3 expression by flow cytometry on CD8⁺ T cells. For all panels, * denotes a p-value < 0.05, two-sided t-test.