Nanotechnology Characterization Laboratory, Cancer Research Technology Program, Frederick National Laboratory for Cancer Research, Frederick, MD, USA.
Methods Mol Biol. 2024;2789:161-169. doi: 10.1007/978-1-0716-3786-9_17.
Nanoparticles are frequently considered in vaccine applications due to their ability to co-deliver multiple antigens and adjuvants to antigen-presenting cells. Some nanoparticles also have intrinsic adjuvant properties that further enhance their ability to stimulate immune cells. The delivery of tumor-specific antigens to antigen-presenting cells (APCs) with subsequent antigenic peptide presentation in the context of class I major histocompatibility complex (MHC-I) molecules represents an essential effort in developing nanotechnology-based cancer vaccines. Experimental models are, therefore, needed to gauge the efficiency of nanotechnology carriers in achieving peptide antigen delivery to APCs and presentation in the context of MHC-I. The assay described herein utilizes a model antigen ovalbumin and model APCs, murine bone marrow-derived dendritic cells. The 25-D1.16 antibody, specific to the ovalbumin (OVA) MHC-I peptide SIINFEKL, recognizes this peptide presented in the context of the murine H2-K class I MHC molecule, allowing the presentation of this antigen on APCs to be detected by flow cytometry after nanoparticle delivery.
纳米颗粒由于能够将多种抗原和佐剂共同递送至抗原呈递细胞而经常被应用于疫苗研究中。一些纳米颗粒本身具有内在的佐剂特性，进一步增强了它们刺激免疫细胞的能力。将肿瘤特异性抗原递送至抗原呈递细胞（APCs），并在 I 类主要组织相容性复合物（MHC-I）分子的背景下呈递抗原肽，这是开发基于纳米技术的癌症疫苗的重要工作。因此，需要实验模型来评估纳米技术载体在将肽抗原递送至 APC 并在 MHC-I 背景下呈递方面的效率。本文所述的测定方法利用模型抗原卵清蛋白和模型 APC 即鼠骨髓来源的树突状细胞。针对卵清蛋白（OVA）MHC-I 肽 SIINFEKL 的 25-D1.16 抗体识别该肽在鼠 H2-K 类 I MHC 分子背景下的呈递，允许在纳米颗粒递送后通过流式细胞术检测 APC 上的抗原呈递。
