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

Figure 1

Specific humoral and cellular immune responses in C57BL/6 mice immunized with hemocyanins in combination with adjuvants. Groups of three female C57BL/6 mice were immunized subcutaneously on days 1 and 16 with one of the following treatments: 50 μg of KLH, CCH, or FLH alone or in combination with 100 μg of alum or 10 μg of QS-21 in a 1 : 1 (vol/vol) ratio with AddaVax, all in a total volume of 100 μl of PBS (vehicle). A serum sample was taken from each animal on day 37 after the second immunization for analysis by indirect ELISA. (a) Total anti-hemocyanin IgG titers in mouse sera. Values are presented as . The anti-hemocyanin IgG titers for each hemocyanin alone were compared by one-way ANOVA with the same hemocyanin in combination with an adjuvant. , , and . independent experiments. (b) Anti-hemocyanin IgG subclass antibodies from a pool of mice immunized with each hemocyanin alone. For these comparisons, half of the maximum OD obtained for each of the isotypes with the different treatments was used [28]. Representative experiments in which the IgG1, IgG2a, IgG2b, and IgG3 titers from each of the treatments were compared by two-way ANOVA. , , and ; independent experiments. (c) Mice immunized with hemocyanins in combination with alum. (d) Mice immunized with hemocyanins in combination with AddaVax. (e) Mice immunized with hemocyanins in combination with QS-21. (f) DTH footpad reactions induced by intradermal challenge with hemocyanins alone in mice immunized with each hemocyanin alone. The DTH test was performed by applying each hemocyanin treatment to the left feet of the mice and measuring the difference in thickness from the right foot at the start of the experiment (time 0) and after 24, 48, and 72 hours. (g) Mice immunized with hemocyanins in combination with alum. (h) Mice immunized with hemocyanins in combination with AddaVax. (i) Mice immunized with hemocyanins in combination with QS-21. Values are presented as . One-way ANOVA was used to compare the foot thickness of the control group at different measurement time points to those of the other treatments at the same time points. , , and , ; independent experiments.

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