1. Volume 13, Issue 1, Pages 194-202 (January 2006)
Enhancement of Immunity by a DNA Melanoma Vaccine against TRP2 with CCL21 as an Adjuvant 
Tomoki Yamano, Yasufumi Kaneda, Sharon Huang, Suzanne H. Hiramatsu, Dave S.B. Hoon 
Molecular Therapy 
Volume 13, Issue 1, Pages (January 2006)
DOI: /j.ymthe
Copyright © 2005 The American Society of Gene Therapy Terms and Conditions

2. FIG. 1
DTH response against rTRP2 after vaccination. Vaccine protocols: (vertical stripes) mock, (empty bar) TRP2, (horizontal stripes) T-C, and (filled bar) C-T. One week after vaccination, DTH was measured 24 and 48 h after rTRP2 injection. Each bar represents the mean ± SD of DTH from four pooled independent experiments (n = 5 mice/experiment). One DTH unit = 0.01 mm. C-T vs other vaccines, P < 0.001.
Molecular Therapy  , DOI: ( /j.ymthe )
Copyright © 2005 The American Society of Gene Therapy Terms and Conditions

3. FIG. 2
Activation of cytokine responses by vaccines. Cytokine production by spleen cells after stimulation with rTRP2 is shown. Spleen cells (1 week after second vaccination) were stimulated (□) or not stimulated (▪) with rTRP2 for 48 h and then specific cytokine production was measured by ELISA. Data show the means ± SE of a representative experiment (n = 5). IFN-γ production: C-T induced significantly higher IFN-γ production than all other vaccine protocols.
Molecular Therapy  , DOI: ( /j.ymthe )
Copyright © 2005 The American Society of Gene Therapy Terms and Conditions

4. FIG. 3
IFN-γ production by spleen cells after coculture with B16 or EL4 cells. Spleen cells were cocultured with B16 (□) or EL4 (▪) cells for 24 h, then IFN-γ production was measured by ELISA. Data represent the means ± SE from pooled data of three independent experiments (n = 5 mice/experiment). C-T spleen cells cocultured with B16 vs other vaccine protocols, P < 0.05. There was no significant difference in IFN-γ production against EL4 among vaccines.
Molecular Therapy  , DOI: ( /j.ymthe )
Copyright © 2005 The American Society of Gene Therapy Terms and Conditions

5. FIG. 4
Prophylactic vaccine protection against B16 melanoma cell challenge. The percentage of tumor rejection is shown; numbers in parentheses indicate the numbers of mice rejecting tumor/total mice. The results of five independent experiments were pooled.
Molecular Therapy  , DOI: ( /j.ymthe )
Copyright © 2005 The American Society of Gene Therapy Terms and Conditions

6. FIG. 5
Timing effect of CCL21 administration. Cytokine production in vitro by spleen cells is shown. CCL21 was administered simultaneous with (C-T(S)), 24 h before (C-T), or 3 days before (C-T(D3)) TRP2 DNA vaccine or C-T was given once (C-T (once)). One week after the second vaccine, spleen cells were harvested and stimulated with (□) or without (▪) rTRP2 for 48 h and assessed for cytokine production. Data show the means ± SE of a representative experiment (n = 5) of three independent experiments.
Molecular Therapy  , DOI: ( /j.ymthe )
Copyright © 2005 The American Society of Gene Therapy Terms and Conditions

7. FIG. 6
Immunostaining analysis of vaccine site draining lymph nodes after TRP2 vaccine alone (T) or CCL21+ TRP2 vaccine (C-T). Immunostaining of lymph nodes at day 7 (B) vs day 1 (A) shows CD11c+ (brown) and CCL21+ cells (purple) increased (a, b), CCR7+ cells (purple) of lymph nodes (c, d), and LYVE+ cells (purple) of lymph nodes (e, f).
Molecular Therapy  , DOI: ( /j.ymthe )
Copyright © 2005 The American Society of Gene Therapy Terms and Conditions