Article Figures & Data
Figures
- Figure 1
Protection of mice immunized with hTRP-2 from syngeneic tumor challenge. Groups of 3-5 C57BL/6 mice (wild type) and mice of the same background deficient in IL4 (IL4-/-) or IFN-gamma (IFN-gamma-/-) were immunized weekly with hTRP-2. Intravenous tumor challenge was performed 7 days after the last immunization and lung metastases were counted 14 days after challenge. Error bars represent standard error of the mean.
- Figure 2
Coat depigmentation in mice immunized with hTRP-2. Groups of 12-19 C57BL/6 mice (wild type) and mice of the same background deficient in IL4 (IL4-/-) or IFN-gamma (IFN-gamma-/-) were immunized with hTRP-2 and observed for the appearance of white hair. (A) The appearance of representative mice observed for 70 days after immunization- left to right: wild type, IL4-/-, IFN-gamma-/-. (B) The number of depigmented abdominal quadrants measured over time. IL4-/- mice depigmented most quickly, but wild type animals eventually achieved the same degree of depigmentation.
- Figure 3
Effect of repletion with recombinant murine IFN-gamma on tumor immunity in IFN-gamma-/- animals. Groups of 5-7 wild type and IFN-gamma-/- mice were immunized with hTRP-2. The IFN-gamma-/- mice were divided into three groups: One group received no repletion, one group was repleted with recombinant murine IFN-gamma from the first day of immunization through the end of the tumor challenge (days 1-56), and one group was repleted with recombinant murine IFN-gamma during the effector phase only (days 41-56). Syngeneic tumor challenge with intravenous B16 melanoma was performed and lung metastases were counted twelve days after tumor injection.
- Figure 4
Cytokine release by CD4+ cell lines generated from C57BL/6 mice immunized with hTRP-2 DNA. Five days after the last immunization, CD4+ T lymphocytes were isolated from the spleen and draining inguinal lymph nodes by magnetic bead cell sorting (MACS) and co-cultured with naive irradiated splenocytes with the addition of hTRP-2 237-256 peptide. Following three weekly rounds of in vitro stimulation, cells were stimulated with plate-bound anti-CD3 and anti-CD28 for 6 hours, in the presence of Brefeldin A for the last 2 hours. Flow cytometry was then performed on the cells in order to analyze intracellular cytokines.
- Figure 5
Effect of cytokines on the antibody response to hgp75/TRP-1. Groups of three C57BL/6 mice were immunized five times with hgp75/TRP-1 preceded by treatment with various murine cytokine DNA constructs or control (null) plasmid. Sera were analyzed for the presence of antibodies detecting hgp75/TRP-1 in an immunoprecipitation-Western blot assay. Results are representative of all three mice per group (GM = GM-CSF).
- Figure 6
Anti-hgp75/TRP-1 antibody titers in wild type (129/SvEv) or IFN-gamma receptor deficient (IFN-gammaR-/-) mice. Groups of five mice were immunized with hgp75/TRP-1 weekly for five weeks. Sera were collected one week following the final immunization and used in a B16 cell-based ELISA at serial dilutions to determine titer (last dilution at which absorbance was above background). The horizontal lines represent median titer. Results were confirmed using immunoprecipitation-Western blot assays with serial dilutions of sera.
Volume 1, Issue 1
