Nanotechnology Characterization Laboratory, Cancer Research Technology Program, Frederick National Laboratory for Cancer Research, Frederick, MD, USA.
Methods Mol Biol. 2024;2789:171-184. doi: 10.1007/978-1-0716-3786-9_18.
The assessment of antigen presentation by dendritic cells and subsequent antigen-dependent activation of T lymphocytes is a critical step underlying the efficacy of nanoparticle-based therapeutic vaccines. Since nanoparticle physicochemical properties determine their interactions with the immune system, the early stages of nanotechnology-based vaccine development commonly involve optimizing the particles' properties to create a formulation with desired stability, antigen release, targeting of desired cell populations, and efficacy. To accelerate this process, in vitro models suitable for the rapid assessment of a novel vaccine candidate's efficacy are highly desirable. One such model is described in this protocol. Herein, nanoparticles are formulated to deliver a model antigen, SIINFEKL (OVA), the immunodominant class I peptide derived from ovalbumin. These nanoparticles are added to the culture of murine bone marrow-derived dendritic cells, which are subsequently co-incubated with CD8 T cells from OT-I transgenic mice. The efficient antigen presentation by dendritic cells results in the antigen-dependent proliferation of CD8 T cells, which is detected by flow cytometry.
树突状细胞对抗原的呈递以及随后抗原依赖性 T 淋巴细胞的激活是基于纳米颗粒的治疗性疫苗功效的关键步骤。由于纳米颗粒的物理化学性质决定了它们与免疫系统的相互作用，因此基于纳米技术的疫苗开发的早期阶段通常涉及优化颗粒的特性，以创建具有所需稳定性、抗原释放、针对目标细胞群体和功效的制剂。为了加速这一过程，非常需要适合快速评估新型疫苗候选物功效的体外模型。本方案中描述了这样一种模型。在此，将纳米颗粒制成可递呈模型抗原 SIINFEKL（OVA），即源自卵清蛋白的免疫优势 I 类肽。将这些纳米颗粒添加到来自 OT-I 转基因小鼠的 CD8 T 细胞与鼠骨髓来源的树突状细胞的共培养物中。树突状细胞的有效抗原呈递导致 CD8 T 细胞的抗原依赖性增殖，通过流式细胞术检测到这一点。
