Adjuvant effect of dendritic cells activator Imiquimod in genetic immunization with HIV-1 p55 Gag.

Journal of immunological techniques in infectious diseases Pub Date : 2023-01-01 Epub Date: 2023-01-10
Janaina M Alves, Mikhail Inyushin, Vassiliy Tsytsarev, Joshua A Roldan-Kalil, Eric Miranda-Valentin, Gerónimo Maldonado-Martinez, Karla M Ramos-Feliciano, Robert Hunter-Mellado
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Abstract

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 a 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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树突状细胞激活剂咪喹莫特在 HIV-1 p55 Gag 基因免疫中的辅助作用
树突状细胞(DC)是重要的抗原递呈细胞,具有诱导和维持 T 细胞免疫或在过度免疫时减弱 T 细胞免疫的能力。进一步激活树突状细胞可能有助于疫苗接种。咪喹莫特是Toll样受体(TLR7)的特异性激动剂,而Toll样受体主要位于DC上。为了研究在小鼠模型中直流刺激对 HIV-1 p55 gag DNA 疫苗有效性的影响,我们使用了 25、50 和 100 nM 的咪喹莫特作为佐剂。随后,我们使用 Western 印迹分析法对免疫后 p55 蛋白的生成进行了定量分析。为了确定T细胞免疫反应的特征,分别通过ELIspot检测法和ELISA法测定了分泌IFN-γ细胞的频率以及IFN-γ和IL-4的产生。研究发现,低浓度的咪喹莫特能有效刺激 Gag 的产生和 T 细胞免疫反应的强度,而高浓度的咪喹莫特则会降低疫苗接种效果。我们的研究结果表明,咪喹莫特的佐剂效应取决于浓度。使用咪喹莫特可能有助于研究DC与T细胞的通讯,包括可能诱导免疫耐受。
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