Hong Enping, Dobrovolskaia Marina A.
Nanotechnology Characterization Lab, Cancer Research Technology Program, Frederick National Laboratory for Cancer Research sponsored by the National Cancer Institute, Frederick, MD 21702
Nanoparticles have been extensively tested as therapeutic vaccine delivery vehicles for the treatment of cancer. A key effort in this area is the delivery of antigens to antigen-presenting cells (APCs), such that tumor-specific peptides are presented in the context of class I major histocompatibility complex (MHC) molecules. The purpose of this protocol is to determine the ability of nanoparticles to deliver antigen to APCs, such that an antigenic class I peptide sequence is properly presented in the context of MHC. For the purposes of this assay, the model antigen ovalbumin (OVA) is used in the context of murine cells. The protocol requires the synthesis of nanoparticles that deliver SIINFEKL (OVA), the immunodominant class I peptide derived from OVA, either in its peptide form or as part of a larger molecule (such as the whole OVA protein). The 25-D1.16 antibody, which binds to SIINFEKL presented in the context of the mouse H2-K MHC molecule [1], is used to detect antigen presentation on APCs after nanoparticle delivery. The results of this protocol may be used to infer the ability of the nanoparticle system to deliver other similar antigens.
纳米颗粒已被广泛测试作为治疗癌症的治疗性疫苗递送载体。该领域的一项关键工作是将抗原递送至抗原呈递细胞（APC），从而使肿瘤特异性肽在I类主要组织相容性复合体（MHC）分子的背景下呈递。本方案的目的是确定纳米颗粒将抗原递送至APC的能力，以便抗原性I类肽序列在MHC的背景下正确呈递。为了该测定的目的，在鼠细胞的背景下使用模型抗原卵清蛋白（OVA）。该方案需要合成递送SIINFEKL（OVA）的纳米颗粒，SIINFEKL是源自OVA的免疫显性I类肽，其可以是肽形式或作为更大分子（如整个OVA蛋白）的一部分。25-D1.16抗体与在小鼠H2-K MHC分子背景下呈递的SIINFEKL结合[1]，用于在纳米颗粒递送后检测APC上的抗原呈递。本方案的结果可用于推断纳米颗粒系统递送其他类似抗原的能力。
