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. Author manuscript; available in PMC: 2018 Dec 28.
Published in final edited form as: Cancer Res. 2014 Sep 9;74(21):5989–5998. doi: 10.1158/0008-5472.CAN-13-2042

Figure 5. HMGN1-gp100 DNA vaccination promoted a Th1-polarized anti-gp100 immune response in vivo.

Figure 5

C57BL/6 mice (7-week old, female, 5 mice/group) were DNA vaccinated with indicated plasmids via gene gun once a week for 3 consecutive weeks. Five days after the last vaccination, single cell suspensions of spleens and draining (inguinal) lymph nodes of vaccinated mouse were stimulated in vitro with 1 µM gp100(25–33) peptide for 6 h at 37°C in humidified air containing 5% CO2, with the addition of GolgiPlug™ for the last 4 h of culture. The cells were immunostained with fluorophore-conjugated antibodies against mouse CD3 and CD8, permeabilized, and followed by staining with fluorophore-conjugated anti-mouse IFNγ and anti-IL-13 antibodies as detailed in the Materials and Methods section. The data were analyzed for the presence of IFNγ+ or IL-13+ CD8 T cells. Shown is the average (mean ± SD) of each group (n=5), representative of one of two separate experiments.