Alves Janaina M, Inyushin Mikhail, Tsytsarev Vassiliy, Roldan-Kalil Joshua A, Miranda-Valentin Eric, Maldonado-Martinez Gerónimo, Ramos-Feliciano Karla M, Hunter-Mellado Robert
Universidad Central del Caribe School of Medicine, Bayamón, Puerto Rico 00960.
University of Maryland School of Medicine, Baltimore, USA.
J Immunol Tech Infect Dis. 2023;12(1). Epub 2023 Jan 10.
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.
树突状细胞(DC)是重要的抗原呈递细胞,具有诱导和维持T细胞免疫的能力,或在超免疫过程中使其减弱。DC的额外激活可能对疫苗接种有用。咪喹莫特是已知的主要位于DC上的Toll样受体(TLR7)的特异性激动剂。为了在小鼠模型中研究DC刺激对HIV-1 p55 gag DNA疫苗有效性的影响,我们使用25、50和100 nM的咪喹莫特作为佐剂。随后,采用蛋白质免疫印迹分析来定量免疫后p55蛋白的产生。为了表征T细胞免疫反应,分别通过酶联免疫斑点分析和酶联免疫吸附测定来测量分泌IFN-γ的细胞频率以及IFN-γ和IL-4的产生。发现低浓度的咪喹莫特能有效刺激Gag的产生以及T细胞免疫反应的强度,而高浓度则会降低疫苗接种效果。我们的结果表明,咪喹莫特的佐剂作用取决于浓度。使用咪喹莫特可能有助于研究DC与T细胞之间的通讯,包括可能诱导免疫耐受。