Figure 5.

Absence of NKT-cell anergy following repeated therapeutic immunisations with I-αGC-LM. BALB/c mice were challenged with 4T1 tumour cells and immunised therapeutically with saline, LM-Mb and I-αGC-LM-Mb as described in Figure 1. Fifteen days after the first immunisation, splenocytes were obtained and stained for NKT cells using αGC-loaded CD1d tetramers (0 h time point). In parallel, a group of mice receiving the same treatment was injected i.p. with free αGC and splenocytes were stained for NKT cells after 12 and 72 h. (A and B) Representative dot plots depicting staining with anti-TCRβ and αGC-loaded CD1d tetramers are shown in A, and the results using N=3 mice per group are summarised in B. Mean values and s.e. are shown. *P<0.05, **P<0.01 (ANOVA with Sidak post-test).