Figure 1.
CpG AuNP conjugate design schematics.
(a) Design 1, CpG-SH, directly has CpG (black lines) assembled on the AuNP surface. (b) Design 2, CpG-T11-SH, incorporates a poly-T nucleotide spacer (light gray lines) to increase the spacing between CpGs (straight arrows). (c) Design 3, CpG-TEG-T11-SH, adds a triethylene glycol (dark gray dotted line) between the CpG sequence and the nucleotide spacer to allow rotation of the CpG segment (curved arrow).
Figure 2.
DNA content characterization and functional TNFα assays of CpG AuNP conjugated constructs.
(a) CpG AuNP conjugate designs and DNA content of each CpG design on the 15 nm, 30 nm and 80 nm AuNPs (*p<0.05). 80 nm AuNP CpG-SH construct aggregated and thus the DNA content was not measured. (b) TNFα levels from macrophage stimulation by CpG conjugated AuNP designs for the three particle sizes (*p<0.05; **p<0.01). (c) TNFα levels normalized to amount of CpG presented by the nanoparticles (*p<0.05; **p<0.01).
Figure 3.
Cytokine and growth factor secretion following stimulation with free CpG, AuNPs coated with control tmCpG, or AuNPs coated with tmCpG for 15 nm, 30 nm and 80 nm core sizes.
(a) TNFα secretion, (b) IL-6 secretion, and (c) G-CSF secretion (pg/ml) (***p<0.0001; **p<0.01; *p<0.05).
Figure 4.
In vivo anti-tumor effect following intratumor injections on days 0, 4, and 7, as indicated (arrows), with PBS, free CpG or tmCpG- AuNP.
(a) Tumor growth and (b) survival percentage after the first CpG injection (*p<0.05).
Figure 5.
Percent tumor infiltration immune cells.
(a) CD11b+ cells, (b) CD11c+, (c) CD8+ cells, (d) CD4+ cells, and (e) CD11b+Gr-1+ cells.