Fig. 2

In vivo and ex vivo fluorescent images of melanomas injected with tyrosinase or RSV promoter-driven bFGF or FGFR-1 antisense constructs. MGP melanomas, grown as subcutaneous tumors on the dorsal site of nude mice, were injected with tyrosinase or RSV promoter-driven bFGF or FGFR-1 antisense constructs as described in Figure 1. Twenty-four hours following the second injection of plasmid construct, each tumor was injected with 5 µg of Cy5-conjugated CD31 antibody. Two hours later, the tumors were imaged noninvasively with a Cy5 excitation and emission filter. Thereafter, the tumors were injected with 5 µg of Cy7-conjugated S100 antibody, and 2 hours later, the tumors were imaged noninvasively using a Cy7 emission and excitation filter. The three groups of control tumors were MGP melanomas, which received fluorochrome-conjugated S100 and CD31 antibody but no plasmid, MGP melanomas that received DC-Chol liposomes and the two fluorochrome-conjugated antibodies, and tumors that were injected with pREP7 vector and the two cyanine dye-conjugated antibodies. Noninvasive, dynamic fluorescence imaging of the tumors was performed twice a week. All animals were sacrificed when the control tumor-bearing mice reached the maximal allowable size of 2 cm in perpendicular diameter. Examples of pseudocolored, noninvasive fluorescent images of control tumors, captured after 2 weeks of intratumoral antibody injections, are depicted in (A). (Fig. 2A, panels a and d) An MGP melanoma, which received only Cy7-conjugated S100 (panel a, pseudocolored red), and Cy5-conjuagted CD31 antibody (panel d, pseudocolored green). Panels (b) and (e) show an MGP melanoma injected with DC-Chol liposomes and Cy7-S100 (panel b) and Cy5-CD31 antibody (panel e). (Fig. 2A, panels c and f) A tumor that was inoculated with pREP7 vector, mixed with DC-Chol liposomes, followed by injection of Cy7-S100 (panel c) and Cy7-CD31 antibody (panel f). (Fig. 2B, panels a–h) Pseudocolored, noninvasively captured fluorescent images of MGP melanomas 2 weeks after injections with tyrosinase or RSV-promoter-driven bFGF or FGFR-1 antisense constructs and fluorochrome-conjugated S100 and CD31 antibody. A tumor that received Tyr-bFGF antisense construct and Cy7-S100 antibody is displayed in (panel a). A Cy5-CD31 fluorescent image of the same tumor is shown in (panel e). The tumor shown in panels b and f received Tyr-FGFR-1 antisense construct and Cy7-S100 (panel b) and Cy5-CD31 antibody (panel f). Panel c depicts a tumor inoculated with RSV-bFGF antisense construct and Cy7-S100 and Cy5-CD31 antibody (panel g). An MGP melanoma, which received RSV-FGFR-1 antisense-oriented plasmid and Cy7-S100 and Cy5-CD31 antibody, is presented in panels d and h, respectively. Displayed in Fig. 2C, panels a–g are superimposed, pseudocolored fluorescent images of tissue sections, which were prepared from the noninvasively imaged tumors, depicted in Fig. 2A, and imaged in the Cy5 and Cy7 channel. (Panel a) uninjected MGP melanoma; (panel b) MGP melanoma injected with DC-Chol liposomes; (panel c) MGP melanoma inoculated with pREP7 vector; (panel d) tumor inoculated with Tyr-bFGF antisense construct; (panel e) tumor injected with Tyr-FGFR-1 antisense construct; (panel f) MGP melanoma that received RSV-bFGF antisense-oriented vector; and (panel g) tumor injected with RSV-FGFR-1 antisense construct. Melanoma cells are pseudocolored, green, and tumor-interspersing blood vessels, red.