Janaina M. Alves, Mikhail Inyushin, Vassiliy Tsytsarev, Joshua A. Roldan-Kalil, Eric Miranda-Valentin, Gerónimo Maldonado-Martinez, Karla M. Ramos-Feliciano, Robert HunterMellado
Dendritic Cells (DC) are important antigen-presenting cells that have abilities to induce and maintain T-cell immunity, or attenuate it during hyperimmunization. Additional activation of DCs may be useful for vaccination purposes. Imiquimod is known to be a specific agonist of the Toll-Like Receptors (TLR7), which are located mainly on DCs. To study the effect of DC stimulation on the effectiveness of an HIV-1 p55 gag DNA vaccine in mice model, we employed 25, 50 and 100 nM of imiquimod as an adjuvant. Subsequently, western blot analysis was used to quantify p55 protein production after the immunization. To characterize T-cells immune response, both the frequency of IFN-γ-secreting cells and IFN-γ and IL-4 production were measured, via an ELIspot assay and ELISA, respectively. Low concentrations of imiquimod were found to effectively stimulate Gag production and the magnitude of the T- cell immune response, whereas higher concentrations reduced vaccination effects. Our results show that the adjuvant effects of imiquimod depend on concentration. The use of imiquimod may be helpful to study DC to T cell communication, including possible induction of immunotolerance.
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