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. Author manuscript; available in PMC: 2013 Sep 15.
Published in final edited form as: Cancer Res. 2012 Aug 3;72(18):4642–4651. doi: 10.1158/0008-5472.CAN-11-3775

Figure 2. B-16-OVA melanoma induces anergy in antigen specific CD4+ T cells in the tumor DLN.

Figure 2

A–D. OT-II mice were challenged with 5×105 B16-OVA cells. Ten to twelve days post tumor challenge, CD4+ T cells were isolated from the DLN and distal non-DLN and stimulated with OVA323–339 peptide-loaded splenocytes. IL-2, IFNγ, IL-17 and IL-4 production were measured by ELISA. Results are presented as mean+SEM from 3–5 mice. *P<0.05; **P<0.01. E. Cell proliferation was also measured in cells stimulated with anti-CD3 and anti-CD28 by BrdU incorporation. Results from are presented as mean±SEM from 4 independent experiments. F. CD4+ T cells isolated from DLN and non-DLN of the B16-OVA challenged OT-II mice (106 B16 cells per mouse) were lysed after isolation for extraction of RNA. cDNA was synthesized and levels of anergy-associated genes transcripts were measured by real-time PCR. Results, expressed as fold induction of the expression of anergy-associated genes in T cells from DLN and non-DLN compared to T cells from control non-tumor bearing mice, are presented as mean±SEM from 3 to 4 independent experiments.