Vaccines containing ribavirin and methods of use thereof
A technology of ribavirin and vaccine, which is applied in the field of compositions for enhancing the effect of vaccines in animals and humans, and can solve problems such as being in the early stage
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Embodiment 1
[0035]The following assays can be used with any ribavirin derivative or combination of ribavirin derivatives to determine the degree of adjuvant activity of a particular composition. In the first set of experiments, each group of 3- 5 Balb / c mice (BK Universal, Uppsala, Sweden). rNS3 was dissolved in phosphate buffered saline (PBS) or PBS containing 1 mg ribavirin (from ICN, Costa Mesa, CA). The mice were injected with a total volume of 100 μl per injection.
[0036] At 2, 4 and 6 weeks after the first i.p. immunization, all mice were sampled by retro-orbital bleeding. Serum samples were collected and analyzed for the presence of antibodies against rNS3. To determine antibody titers, an enzyme immunoassay (EIA) was performed. (See eg Hultgren et al., J Gen Virol. 79:2381-91 (1998) and Hultgren et al., Clin. Diagn. Lab. Immunol. 4:630-632 (1997)). The highest serum dilution at which the optical density at 405nm exceeds twice that of non-immunized mice was recorded as the a...
Embodiment 2
[0047] To determine the dose of ribavirin sufficient to provide an adjuvant effect, the following experiments were performed. In the first group of experiments, each group of mice (3 mice in each group) was immunized with 20 μg rNS3 alone or a mixture of 20 μg rNS3 and 0.1 mg, 1 mg or 10 mg ribavirin. Antibody levels against the antigen were then determined by EIA. Plot the mean endpoint titer curves for weeks 1 and 3, as figure 2 shown. The adjuvant effect provided by ribavirin was found to have different kinetics depending on the dose of ribavirin provided. For example, low doses (<1 mg) ribavirin were found to enhance antibody levels even at week 1 but not week 3, whereas higher doses (1-10 mg) were found to enhance antibody levels at week 3.
[0048] A second set of experiments was also performed. In these experiments, groups of mice were injected with vaccine compositions comprising varying amounts of ribavirin and rNS3, and IgG responses in these animals were monito...
Embodiment 3
[0054] This assay can be used with any ribavirin derivative or combination of ribavirin derivatives to determine the extent to which a particular vaccine formulation modulates the cellular immune response. To determine CD4 + T cell responses to ribavirin-containing vaccines. Groups of mice were immunized with 100 μg rNS3 in PBS or 100 μg rNS3 and 1 mg ribavirin in PBS s.c. Mice were sacrificed 10 days after immunization, and their lymph nodes were harvested and drained. Then an in vitro memory test was performed. (See eg Hultgren et al., J Gen Virol. 79:2381-91 (1998) and Hultgren et al., Clin. Diagn. Lab. Jmnmnol. 4:630-632 (1997)). [ 3 H] thymidine assay for CD4 + T cell proliferation.
[0055] Such as image 3 As shown, mice immunized with 100 μg rNS3 mixed with 1 mg ribavirin had a much greater T cell proliferative response than mice immunized with 100 μg rNS3 in PBS. These data provide further evidence that ribavirin enhances or promotes cellular immune responses (...
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