Immunotherapeutic Potential of Mollusk Hemocyanins in Combination with Human Vaccine Adjuvants in Murine Models of Oral Cancer

Figure 4

Humoral and cellular immune responses of the mice from the prophylactic bioassay utilizing the oral cancer orthotopic model consisting of C3H/He mice challenged with the AT-84 E7 Luc cell line. Groups of five female C3H/He mice were immunized with one of the following treatments: 100 μg of KLH or FLH, 50 μg of KLH or FLH plus 10 μg of QS-21, and 10 μg of QS-21 or 100 μl of PBS. At 10 and 25 days, serum samples were taken to determine the titers of IgG subclasses by indirect ELISA. DTH determination was performed on day 15 of the bioassay. (a) Total anti-hemocyanin IgG titers in mouse sera. Data are shown as . Anti-hemocyanin IgG titers for each hemocyanin alone were compared by one-way ANOVA with the same hemocyanin in combination with QS-21 at day 10 and at day 21. and . Representative experiment with mice per group. (b) Anti-hemocyanin IgG subclass antibodies at day 10. Half of the maximum OD obtained for each of the isotypes with the different treatments was used for comparison [28]. (c) Anti-hemocyanin IgG subclass antibodies at day 25. Half of the maximum OD obtained for each of the isotypes with the different treatments was used for comparison [28]. IgG1 was compared against IgG2a, IgG2b, and IgG3 for each treatment. , , and , two-way ANOVA. (d) Delayed hypersensitivity reaction. This assay was performed by applying the corresponding hemocyanin for each treatment group to the left paw of the mouse and subsequently measuring the difference in thickness from the right paw after 24, 48, and 72 hours. Data are shown as of a representative experiment with mice per group. The foot thickness in the control group was compared at different measurement time points with those in the other treatments at the same time points. , , , and , analyzed by one-way ANOVA.

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